| 著作名稱： | Biotransformation of celastrol to a novel, well-soluble, low-toxic and antioxidant celastrol-29-O-beta-glucoside by Bacillus glycosyltransferases |
| 年度： | 2021 |
| 類別： |
期刊論文
Journal of Bioscience and Bioengineering
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| 摘要： | Celastrol is a quinone-methide triterpenoid isolated from the root extracts of Tripterygium wilfordii (Thunder god vine). Although celastrol possesses multiple bioactivities, the potent toxicity and rare solubility in water hinder its clinical application. Biotransformation of celastrol using either whole cells or purified enzymes to form less toxic and more soluble derivatives has been proven difficult due to its potent antibiotic and enzyme-conjugation property. The present study evaluated biotransformation of celastrol by four glycosyltransferases from Bacillus species and found one glycosyltransferase (BsGT110) from Bacillus subtilis with significant activity toward celastrol. The biotransformation metabolite was purified and identified as celastrol-29-O-β-glucoside by mass and nuclear magnetic resonance spectroscopy. Celastrol-29-O-β-glucoside showed over 53-fold higher water solubility than celastrol, while maintained 50% of the free radical scavenging activity of celastrol. When using zebrafish as the in vivo animal model, celastrol-29-O-β-glucoside exhibited 50-fold less toxicity than celastrol. To our knowledge, the present study is not only the first report describing the biotransformation of celastrol, but also the first one detailing a new compound, celastrol-29-O-β-glucoside, that is generated in the biotransformation process. Moreover, celastrol-29-O-β-glucoside may serve as a potential candidate in the future medicine application due to its higher water solubility and lower toxicity. |
| 關鍵字： | Bacillus; Biotransformation; Celastrol; Glycosyltransferase; Triterpenoid. |
| 著作名稱： | Production of a new triterpenoid disaccharide saponin from sequential glycosylation of ganoderic acid A by two novel Bacillus glycosyltransferases |
| 年度： | 2021 |
| 類別： |
期刊論文
Bioscience, Biotechnology, and Biochemistry
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| 摘要： | Ganoderic acid A (GAA) is a lanostane-type triterpenoid, isolated from medicinal fungus Ganoderma lucidum, and possesses multiple bioactivities. In the present study, GAA was sequentially biotransformed by 2 recently discovered Bacillus glycosyltransferases (GT), BtGT_16345 and BsGT110, and the final product was purified and identified as a new compound, GAA-15,26-O-β-diglucoside, which showed 1024-fold aqueous solubility than GAA. |
| 關鍵字： | ganoderic acid A, glycosyltransferase, triterpenoid, glucosides, saponin |
| 著作名稱： | One-Pot Bi-Enzymatic Cascade Synthesis of Novel Ganoderma Triterpenoid Saponins |
| 年度： | 2021 |
| 類別： |
期刊論文
Catalysts
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| 摘要： | Ganoderma lucidum is a medicinal fungus whose numerous triterpenoids are its main bioactive constituents. Although hundreds of Ganoderma triterpenoids have been identified, Ganoderma triterpenoid glycosides, also named triterpenoid saponins, have been rarely found. Ganoderic acid A (GAA), a major Ganoderma triterpenoid, was synthetically cascaded to form GAA-15-O-β-glucopyranoside (GAA-15-G) by glycosyltransferase (BtGT_16345) from Bacillus thuringiensis GA A07 and subsequently biotransformed into a series of GAA glucosides by cyclodextrin glucanotransferase (Toruzyme® 3.0 L) from Thermoanaerobacter sp. The optimal reaction conditions for the second-step biotransformation of GAA-15-G were found to be 20% of maltose; pH 5; 60 °C. A series of GAA glucosides (GAA-G2, GAA-G3, and GAA-G4) could be purified with preparative high-performance liquid chromatography (HPLC) and identified by mass and nucleic magnetic resonance (NMR) spectral analysis. The major product, GAA-15-O-[α-glucopyranosyl-(1→4)-β-glucopyranoside] (GAA-G2), showed over 4554-fold higher aqueous solubility than GAA. The present study demonstrated that multiple Ganoderma triterpenoid saponins could be produced by sequential actions of BtGT_16345 and Toruzyme®, and the synthetic strategy that we proposed might be applied to many other Ganoderma triterpenoids to produce numerous novel Ganoderma triterpenoid saponins in the future. |
| 關鍵字： | Ganoderma lucidum; ganoderic acid A; triterpenoid; glycosyltransferase; cyclodextrin glucanotransferase; saponin |
| 著作名稱： | Production of new isoflavone diglucosides from glycosylation of 8-hydroxydaidzein by Deinococcus geothermalis amylosucrase |
| 年度： | 2021 |
| 類別： |
期刊論文
Fermentation
|
| 摘要： | first_pagesettings
Open AccessArticle
Production of New Isoflavone Diglucosides from Glycosylation of 8-Hydroxydaidzein by Deinococcus geothermalis Amylosucrase
by Chien-Min Chiang 1,†ORCID,Tzi-Yuan Wang 2,†ORCID,Jiumn-Yih Wu 3,†,Yun-Rong Zhang 4,Shu-Yuan Lin 4 andTe-Sheng Chang 4,*ORCID
1
Department of Pharmacy, Chia Nan University of Pharmacy and Science, No. 60, Sec. 1, Erh-Jen Rd., Jen-Te District, Tainan 71710, Taiwan
2
Biodiversity Research Center, Academia Sinica, Taipei 11529, Taiwan
3
Department of Food Science, National Quemoy University, Kinmen County 892, Taiwan
4
Department of Biological Sciences and Technology, National University of Tainan, Tainan 70005, Taiwan
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this manuscript.
Academic Editors: Clemencia Chaves-López and Alexander Rapoport
Fermentation 2021, 7(4), 232; https://doi.org/10.3390/fermentation7040232
Received: 23 August 2021 / Revised: 1 October 2021 / Accepted: 14 October 2021 / Published: 16 October 2021
(This article belongs to the Special Issue Fermentation and Bioactive Metabolites 3.0)
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Abstract
8-Hydroxydaidzein (8-OHDe) is a non-natural isoflavone polyphenol isolated from fermented soybean foods. 8-OHDe exhibits a wide range of pharmaceutical activities. However, both the poor solubility and instability of 8-OHDe limit its applications. To resolve the limitations of 8-OHDe, Deinococcus geothermalis amylosucrase (DgAS) has previously been used to glycosylate 8-OHDe to produce soluble and stable 8-OHDe-7-O-α-glucopyranoside (8-OHDe-7-G) in a 0.5 h reaction time. In this study, we aimed to use DgAS and an extended reaction time to produce 8-OHDe diglucosides. At least three 8-OHDe derivatives were produced after a 24 h reaction time, and two major products were successfully purified and identified as new compounds: 8-OHDe-7-O-[α-glucopyranosyl-(1→6)-α-glucopyranoside] (8-OHDe-7-G2) and 8-OHDe-7,4′-O-α-diglucopyranoside (8-OHDe-7-G-4′-G). 8-OHDe-7-G-4′-G showed a 4619-fold greater aqueous solubility than 8-OHDe. In addition, over 92% of the 8-OHDe diglucosides were stable after 96 h, while only 10% of the 8-OHDe could be detected after being subjected to the same conditions. The two stable 8-OHDe diglucoside derivatives have the potential for pharmacological usage in the future. |
| 關鍵字： | amylosucrase; Deinococcus geothermalis; 8-hydroxydaidzein; isoflavone; glycosylation |
| 著作名稱： | Enzymatic synthesis of novel vitexin glucosides |
| 年度： | 2021 |
| 類別： |
期刊論文
Molecules
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| 摘要： | Vitexin is a C-glucoside flavone that exhibits a wide range of pharmaceutical activities. However, the poor solubility of vitexin limits its applications. To resolve this limitation, two glycoside hydrolases (GHs) and four glycosyltransferases (GTs) were assayed for glycosylation activity toward vitexin. The results showed that BtGT_16345 from the Bacillus thuringiensis GA A07 strain possessed the highest glycosylation activity, catalyzing the conversion of vitexin into new compounds, vitexin-4′-O-β-glucoside (1) and vitexin-5-O-β-glucoside (2), which showed greater aqueous solubility than vitexin. To our knowledge, this is the first report of vitexin glycosylation. Based on the multiple bioactivities of vitexin, the two highly soluble vitexin derivatives might have high potential for pharmacological usage in the future. |
| 關鍵字： | glycoside hydrolase; glycosyltransferase; glycosylation; vitexin; glucoside |
| 著作名稱： | Glycosylation of ganoderic acid G by Bacillus glycosyltransferases |
| 年度： | 2021 |
| 類別： |
期刊論文
Int J Mol Sci
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| 摘要： | Ganoderma lucidum is a medicinal fungus abundant in triterpenoids, its primary bioactive components. Although numerous Ganoderma triterpenoids have already been identified, rare Ganoderma triterpenoid saponins were recently discovered. To create novel Ganoderma saponins, ganoderic acid G (GAG) was selected for biotransformation using four Bacillus glycosyltransferases (GTs) including BtGT_16345 from the Bacillus thuringiensis GA A07 strain and three GTs (BsGT110, BsUGT398, and BsUGT489) from the Bacillus subtilis ATCC 6633 strain. The results showed that BsUGT489 catalyzed the glycosylation of GAG to GAG-3-o-β-glucoside, while BsGT110 catalyzed the glycosylation of GAG to GAG-26-o-β-glucoside, which showed 54-fold and 97-fold greater aqueous solubility than that of GAG, respectively. To our knowledge, these two GAG saponins are new compounds. The glycosylation specificity of the four Bacillus GTs highlights the possibility of novel Ganoderma triterpenoid saponin production in the future. |
| 關鍵字： | Bacillus; Ganoderma lucidum; glycosyltransferase; saponin; triterpenoid. |
| 著作名稱： | Improving aqueous solubility of natural antioxidant mangiferin through glycosylation by maltogenic amylase from Parageobacillus galactosidasius DSM 18751 |
| 年度： | 2021 |
| 類別： |
期刊論文
Antioxidants
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| 摘要： | Mangiferin is a natural antioxidant C-glucosidic xanthone originally isolated from the Mangifera indica (mango) plant. Mangiferin exhibits a wide range of pharmaceutical activities. However, mangiferins poor solubility limits its applications. To resolve this limitation of mangiferin, enzymatic glycosylation of mangiferin to produce more soluble mangiferin glucosides was evaluated. Herein, the recombinant maltogenic amylase (MA; E.C. 3.2.1.133) from a thermophile Parageobacillus galactosidasius DSM 18751T (PgMA) was cloned into Escherichia coli BL21 (DE3) via the expression plasmid pET-Duet-1. The recombinant PgMA was purified via Ni2+ affinity chromatography. To evaluate its transglycosylation activity, 17 molecules, including mangiferin (as sugar acceptors), belonging to triterpenoids, saponins, flavonoids, and polyphenol glycosides, were assayed with β-CD (as the sugar donor). The results showed that puerarin and mangiferin are suitable sugar acceptors in the transglycosylation reaction. The glycosylation products from mangiferin by PgMA were isolated using preparative high-performance liquid chromatography. Their chemical structures were glucosyl-α-(1→6)-mangiferin and maltosyl-α-(1→6)-mangiferin, determined by mass and nucleic magnetic resonance spectral analysis. The newly identified maltosyl-α-(1→6)-mangiferin showed 5500-fold higher aqueous solubility than that of mangiferin, and both mangiferin glucosides exhibited similar 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activities compared to mangiferin. PgMA is the first MA with glycosylation activity toward mangiferin, meaning mangiferin glucosides have potential future applications. |
| 關鍵字： | Parageobacillus galactosidasius; glucoside; glycosylation; maltogenic amylase; mangiferin. |
| 著作名稱： | Biotransformation of isoflavones daidzein and genistein by recombinant Pichia pastoris expressing membrane-anchoring and reductase fusion chimeric CYP105D7 |
| 年度： | 2016 |
| 類別： |
期刊論文
Journal of Taiwan Chemical Engineerers
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| 摘要： | The biotransformation of soy isoflavones into ortho-hydroxyisoflavones by CYP105D7 from Streptomyces avermitilis MA4680 is investigated through chimeric expression in Pichia pastoris. Using N-terminal fusion with the transmembrane domain of CYP57B3 from Aspergillus oryzae and C-terminal fusion with a P450 reductase from Saccharomyces cerevisiae, CYP105D7 was expressed in the form of reductase fusion and membrane anchoring in P. pastoris. Recombinant P. pastoris expressing the chimera catalyzed the biotransformation of both daidzein and genistein. This is the first study to show the catalyzing activity of CYP105D7 toward genistein. The major product from daidzein was identified as 6-hydroxydaidzein by comparing the results of the ultra-performance liquid chromatography analysis with the authentic standard. The major product from genistein was purified using preparative high-performance liquid chromatography and identified as 3-hydroxygenistein based on nuclear magnetic resonance and mass data. The recombinant P. pastoris produced 6-hydroxydaidzein and 3-hydroxygenistein in a 5-L fermenter, with maximal yields of 7.5 and 15.0 mg/l, respectively. The production of 3-hydroxygenistein was higher than any previously reported in the literature. |
| 關鍵字： | Keywords: hydroxylation, daidzein, genistein, cytochrome P450 monooxygenase, Pichia pastoris, Streptomyces avermitilis? |
| 著作名稱： | Identification of 3’-hydroxygenistein as a potent melanogenesis inhibitor from biotransformation of genistein by recombinant Pichia pastoris. |
| 年度： | 2015 |
| 類別： |
期刊論文
Process Biochemisty
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| 摘要： | A product resulting from the biotransformation of genistein by a recombinant Pichia pastoris was isolated and identified as 3-hydroxygenistein, on the basis of mass, 1H-NMR, and 13C-NMR spectrophotometric analysis. The maximal product concentration and the conversion yield of the biotransformation in a 5 l fermenter were 3.5 mg/l and 14%, respectively. The inhibitory effects of 3-hydroxygenistein on tyrosinase activity were investigated in vitro using mushroom tyrosinase. The results showed that 3-hydroxygenistein potently inhibited tyrosinase activity with an IC50 value of 15.9 M. Furthermore, the inhibitory effects of 3-hydroxygenistein on melanogenesis were also investigated in vitro in cultured B16 melanoma cells, and it was shown that 3-hydroxygenistein dose-dependently inhibited melanogenesis in non-toxic concentrations. In summary, the 3-hydroxygenistein that was produced from genistein by the recombinant yeast was confirmed as a potent tyrosinase inhibitor and inhibited melanogenesis in B16 cells. |
| 關鍵字： | 3-hydroxygenistein, genistein, melanogenesis, tyrosinase, inhibition, Pichia pastoris |
| 著作名稱： | Inhibition of Melanogenesis by Yeast Extracts from Cultivations of Recombinant Pichia pastoris Catalyzing ortho-Hydroxylation of Flavonoids |
| 年度： | 2015 |
| 類別： |
期刊論文
Cur. Pharm. Biotehcnol.
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| 摘要： | The inhibition of melanogenesis by yeast extracts from cultivations of recombinant Pichia pastoris catalyzing ortho-hydroxylation of flavonoids was investigated. The recombinant yeast harbored a fusion gene composed of the CYP57B3 gene from Aspergillus oryzae and a cytochrome reductase gene from Saccharomyces cerevisiae. Ten flavonoids belonging to flavones, flavonols, flavanones, flavanols, and isoflavones were evaluated for biotransformation by the recombinant strain. The results showed that five flavonoids, including the flavone apigenin, the flavanones naringenin and liquiritigenin, and the isoflavones daidzein and genistein, could be biotransformed. The yeast extracts from the five biotransformation fermentations were then evaluated for inhibitory activity on melanogenesis in cultured mouse B16 melanoma cells. Three yeast extracts from biotransformation fermentation feeding with daidzein, genistein, or apigenin showed inhibitory activity on melanogenesis in the B16 cells, while the extract from genistein biotransformation exhibited the highest activity. The yeast extract from genistein biotransformation also showed inhibitory activity on cellular tyrosinase activity in the B16 cells. The present study shows a CYP with multiple flavonoid substrates for the first time and highlights the usage of yeast extracts from cultivations of the recombinant yeast catalyzing flavonoids’ biotransformation in the development of skin-whitening agents. |
| 關鍵字： | biotransformation; apigenin; daidzein; genistein; naringenin; liquiritigenin; P450; Pichia pastoris |
| 著作名稱： | Isolation, bioactivity, and production of ortho-hydroxydaidzein and ortho-hydroxygenistein. |
| 年度： | 2014 |
| 類別： |
期刊論文
Int. J. Mol. Sci.
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| 摘要： | Daidzeinandgenisteinaretwomajorcomponentsofsoyisoflavones.Theyexistabundantlyinplantsandpossessmultiplebioactivities.Incontrast,ortho-hydroxydaidzein(OHD)andortho-hydroxygenistein(OHG),including6-hydroxydaidzein(6-OHD),
8-hydroxydaidzein(8-OHD),3-hydroxydaidzein(3-OHD),6-hydroxygenistein(6-OHG),8-hydroxygenistein(8-OHG),and3-hydroxygenistein(3-OHG),arerarelyfoundinplants.Instead,theyareusuallyisolatedfromfermentedsoybeanfoodsormicrobialfermentationbrothfeedingwithsoybeanmeal.Accordingly,thebioactivityofOHDandOHGhasbeeninvestigatedlesscomparedtothatofsoyisoflavones.Recently,OHDandOHGwereproducedbygeneticallyengineeringmicroorganismsthroughgenecloningofcytochromeP450(CYP)enzymesystems.ThissuccessopensupbioactivityinvestigationandindustrialapplicationsofOHDandOHGinthefuture.Thisarticlereviewsisolation
ofOHDandOHGfromnon-syntheticsourcesandproductionofthecompoundsbygeneticallymodifiedmicroorganisms.Severalbioactivities,suchasanticancerandantimelanogenesis-relatedactivities,ofOHDandOHG,arealsodiscussed.
|
| 關鍵字： | soyisoflavones;daidzein;genistein;hydroxylation;ortho-hydroxydaidzein;ortho-hydroxygenistein;bioactivity;isolation;production;cancer;melanogenesis |
| 著作名稱： | Production of ortho-hydroxydaidzein derivatives by a recombinant strain of Pichia pastoris harboring a cytochrome P450 fusion gene. |
| 年度： | 2013 |
| 類別： |
期刊論文
Process Biochemisty
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| 摘要： | CYP57B3 from Aspergillus oryzae was recently discovered to catalyze the ortho-hydroxylation of the soyisoflavone genistein. In the present study, the gene encoding CYP57B3 was fused with the reductase domain of the CYP102A1 gene (BM3R) from Bacillus megaterium, and recombinant Pichia pastoris harboring the P450 fusion gene was evaluated for its ability to produce ortho-hydroxydaidzein derivatives from daidzein. The results showed that 8-hydroxydaidzein (8-OHDe), 3-hydroxydaidzein (3-OHDe), and 6-hydroxydaidzein (6-OHDe) were produced during fermentation with a maximal conversion of 2.4, 0.9, and 36.3 %, respectively. The maximal production concentration of 6-OHDe by the recombinant strain was 9.1 mg/l. To our knowledge, both the maximal production concentration and the conversion efficiency of 6-OHDe from daidzein in the present study are the highest values reported in the literatures to date. The present study is also the first to demonstrate production of ortho-hydroxydaidzein derivatives using a fusion fungus P450. |
| 關鍵字： | hydroxydaidzein, daidzein, Aspergillus oryzae, cytochrome P450 monooxygenase, Pichia pastoris |
| 著作名稱： | Melanogenesis inhibition by homoisoflavanone sappanone A from Caesalpinia sappan. |
| 年度： | 2012 |
| 類別： |
期刊論文
Int. J. Mol. Sci.
|
| 摘要： | Abstract: Homoisoflavanone, sappanone A, was isolated from Caesalpinia sappan and
proven to dose-dependently inhibit both melanogenesis and cellular tyrosinase activity via
repressing tyrosinase gene expression in mouse B16 melanoma cells. To our knowledge,
sappanone A is the first homoisoflavanone to be discovered with melanogenesis inhibitory
activity. Our results give a new impetus to the future search for other homoisoflavanone
melanogenesis inhibitors |
| 關鍵字： | Keywords: Caesalpinia sappan; inhibition; melanogenesis; sappanone A; tyrosinase |
| 著作名稱： | Natural Melanogenesis Inhibitors Acting Through the Down-Regulation of Tyrosinase Activity. |
| 年度： | 2012 |
| 類別： |
期刊論文
Materials
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| 摘要： | Abstract: Melanogenesis is a biosynthetic pathway for the formation of the pigment melanin in human skin. A key enzyme, tyrosinase, catalyzes the first and only rate-limiting steps in melanogenesis, and the down-regulation of enzyme activity is the most reported method for the inhibition of melanogenesis. Because of the cosmetically important issue of hyperpigmentation, there is a big demand for melanogenesis inhibitors. This encourages researchers to seek potent melanogenesis inhibitors for cosmetic uses. This article reviews melanogenesis inhibitors that have been recently discovered from natural sources. The reaction mechanisms of the inhibitors on tyrosinase activity are also discussed. |
| 關鍵字： | Keywords: inhibitors; melanogenesis; tyrosinase |
| 著作名稱： | Chang TS*, Chen CT (2012) Inhibitory Effect of Homochlorcyclizine on Melanogenesis in -Melanocyte Stimulating Hormone-Stimulated Mouse B16 Melanoma Cells. Arch. Pharm. Res. 35, 1: 119-127 [SCI Ranking 78 % (185/236) in the field of Pharm. Pharm., IF 1.159 in 2009]. |
| 年度： | 2012 |
| 類別： |
期刊論文
|
| 摘要： | The histamine receptor H1 antagonist homochlorcyclizine (HC) has been widely used as an antihistamine agent for the treatment of allergies. However, the effect of HC on skin pigmentation is not known. In the present study, we investigated the inhibitory effect of HC on melanogenesis in mouse B16 melanoma cells. Our results showed that HC inhibited melanogenesis in either -melanocyte stimulating hormone (-MSH)- or 3-isobutyl-1-methylxanthin (IBMX)-stimulated B16 cells in a dose-dependent manner. Despite the strong inhibition of melanogenesis by HC, it was surprisingly found that HC did not reduce either cellular or melanosomal tyrosinase activity in -MSH-stimulated B16 cells. In addition, HC also did not directly inhibit either murine or mushroom tyrosinase activity in the cell-free system. Moreover, western blotting and reverse-transcription polymerase chain reaction (RT-PCR) analyses respectively confirmed that HC did not downregulate levels of tyrosinase protein and its mRNA in -MSH-stimulated B16 cells. These results clearly demonstrated that HC inhibits melanogenesis of B16 cells by a mechanism other than reduction of the cellular tyrosinase activity. From the present study, HC was proven to be a good candidate as a skin-whitening agent for treatment of skin hyperpigmentation, and this generic drug might be suitable for use in combination with other depigmenting agents due to its unique inhibition mechanism. |
| 關鍵字： | Homochlorcyclizine, Melanogenesis, Tyrosinase, Melanin, Inhibition |
| 著作名稱： | Ding HY, Chang TS, Chiang CM, Li SY, Tseng DY* (2011) Melanogenesis inhibition by a crude extract of Magnolia officinalis. J. Med. Plants Res. 5, 237-244 |
| 年度： | 2011 |
| 類別： |
期刊論文
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| 摘要： | In the present study, we investigated the inhibitory effect of a crude extract from Magnolia officinalis (MOE) on melanogenesis in both mouse B16 melanoma cells and zebrafish. Our results showed that MOE inhibited melanogenesis in either -melanocyte stimulating hormone (-MSH)- or 3-isobutyl-1-methylxanthin (IBMX)-stimulated B16 cells in a dose-dependent manner with an IC50 value of 9.3 g/ml. In addition, MOE also inhibited cellular tyrosinase activity with an IC50 value of 13.4 g/ml while no inhibitory activity was found by MOE against cell-free tyrosinase activity. Moreover, western blotting and real time reverse-transcription polymerase chain reaction (qRT-PCR) analyses respectively confirmed that MOE downregulated levels of tyrosinase protein but not that of its mRNA in -MSH-stimulated B16 cells. These results demonstrated that MOE inhibits melanogenesis of B16 cells by a pre-translational regulation on tyrosinase gene expression. In the other hand, when using zebrafish as a depigmenting assay system, MOE could inhibit both melanogenesis and tyrosinase activity in the in vivo model. From the present study, MOE was proven to be a good candidate as a skin-whitening agent for treatment of skin hyperpigmentation. |
| 關鍵字： | Magnolia officinalis, Melanogenesis, Tyrosinase, Melanin, Inhibition |
| 著作名稱： | Ding HY, Chang TS, Chiang CM, Tai SK* (2011) Inhibitory Effect of a Water Extract from Pemphis acidula on Melanogenesis in Mouse B16 Melanoma Cells. J. Cosmet. Sci. 62, 41-48. |
| 年度： | 2011 |
| 類別： |
期刊論文
|
| 摘要： | The inhibitory effect of a water extract from Pemphis acidula on melanogenesis in mouse B16 melanoma cells was investigated. The results showed that the P. acidula extract (PAE) inhibited melanogenesis in 3-isobutyl-1-methylxanthin (IBMX)-stimulated B16 cells in a dose-dependent manner with an IC50 value of 33.5 g/ml. In addition, PAE also inhibited cellular tyrosinase activity. Moreover, western blot and real time reverse transcriptase polymerase chain reaction (qRT-PCR) analyses respectively confirmed that PAE downregulated levels of tyrosinase protein and its mRNA in IBMX-stimulated B16 cells. These results demonstrated that PAE inhibits melanogenesis of B16 cells by reducing tyrosinase gene expression. From the present study, PAE was proven to be a good candidate as a skin-whitening agent for treatment of skin hyperpigmentation. |
| 關鍵字： | Pemphis acidula, melanogenesis, tyrosinase, melanin, inhibition |
| 著作名稱： | Lin VC, Ding HY, Tsai PC, Wu JY, Lu YH, Chang TS* (2011) In Vitro and In Vivo Melanogenesis Inhibition by Biochanin A from Trifolium pretense. Biosci. Biotechnol. Biochem. 75, 914-918 |
| 年度： | 2011 |
| 類別： |
期刊論文
|
| 摘要： | Our previous study showed that a methanol extract from Trifolium pratense exerted potent inhibitory activity on melanogenesis in mouse B16 melanoma cells. In the present study, the active compound in this Chinese herb extract was isolated and identified as biochanin A by mass spectrum, 1H-NMR, and 13C-NMR analysis. The inhibitory effects of biochanin A on melanogenesis were investigated in vitro in cultured melanoma cells and in vivo in zebrafish and mice. Biochanin A dose-dependently inhibited both melanogenesis and cellular tyrosinase activity in B16 cells and in zebrafish embryos. Application of a cream containing 2% biochanin A twice daily to the skin of mice also increased the skin-whitening index value after 1 week of treatment, and the increase continued for another 2 weeks. Biochanin A was confirmed as a good candidate for use as a skin-whitening agent in the treatment of skin hyperpigmentation disorders. |
| 關鍵字： | biochanin A; inhibition; melanogenesis; Trifolium pretense; tyrosinase |
| 著作名稱： | Lin VC, Ding HY, Kuo SY, Chin LW, Wu JY, Chang TS* (2011) Evaluation of in Vitro and in Vivo Depigmenting Activity of Raspberry Ketone from Rheum officinale. Int. J. Mol. Sci. 12, 4819-4835 |
| 年度： | 2011 |
| 類別： |
期刊論文
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| 摘要： | Abstract: Melanogenesis inhibition by raspberry ketone (RK) from Rheum officinale was investigated both in vitro in cultivated murine B16 melanoma cells and in vivo in zebrafish and mice. In B16 cells, RK inhibited melanogenesis through a post-transcriptional regulation of tyrosinase gene expression, which resulted in down regulation of both cellular tyrosinase activity and the amount of tyrosinase protein, while the level of tyrosinase mRNA transcription was not affected. In zebrafish, RK also inhibited melanogenesis by reduction of tyrosinase activity. In mice, application of a 0.2% or 2% gel preparation of RK applied to mouse skin significantly increased the degree of skin whitening within one week of treatment. In contrast to the widely used flavoring properties of RK in perfumery and cosmetics, the skin-whitening potency of RK has been demonstrated in the present study. Based on our findings reported here, RK would appear to have high potential for use in the cosmetics industry. |
| 關鍵字： | B16 melanoma; tyrosinase; inhibition; zebrafish; melanogenesis; Rheum officinale; raspberry ketone |
| 著作名稱： | Ding HY, Chang TS, Shen HC, Tai SSK* (2011) Murine Tyrosinase Inhibitors from Cynanchum bungei and Evaluation of in Vitro and in Vivo Depigmenting Activity. Experimental. Dermatol. 20, 720-724. |
| 年度： | 2011 |
| 類別： |
期刊論文
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| 摘要： | Abstract- Two natural acetophenone derivatives, 2,5-dihydroxyacetophenone (2,5-DHAP) and 2,6-dihydroxyacetophenone (2,6-DHAP), were purified from Cynanchum bungei and identified as murine tyrosinase inhibitors. Investigation of 2,5-DHAP showed it to be an uncompetitive inhibitor of murine tyrosinase (KI 0.28 mM). 2,5-DHAP strongly inhibited both melanogenesis and cellular tyrosinase activity in vitro in IBMX-stimulated B16 mouse melanoma cells or in vivo in zebrafish and mouse models, but showed no cytotoxicity at the concentrations used. In B16 cells, 2,5-DHAP inhibition was dose-dependent and was four-fold greater than that of arbutin. 2,5-DHAP had no effect on the expression of tyrosinase protein or mRNA, as confirmed by Western blotting and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), respectively. A 2% gel preparation of 2,5-DHAP applied to the skin of mice significantly increased the average skin whitening index (L value), indicating its potential use as a treatment for skin hyperpigmentation in humans. |
| 關鍵字： | Tyrosinase, Inhibitor, Zebrafish, Melanogenesis, Acetophenone, Cynanchum bungei |
| 著作名稱： | Chang TS*, Lin VC (2011) Melanogenesis Inhibitory Activity of Two Generic Drugs: Cinnarizine and Trazodone in Mouse B16 Melanoma Cells. Int. J. Mol. Sci. 12, 8787-8796 (IF 2.279). |
| 年度： | 2011 |
| 類別： |
期刊論文
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| 摘要： | More than 200 generic drugs were screened to identify the inhibitory activity
on melanogenesis in mouse B16 melanoma cells. Cinnarizine and trazodone were
identified as melanogenesis inhibitors. The inhibitory effects of the two drugs on cell
survival, melanogenesis, and tyrosinase activity were investigated. The results showed that
both cinnarizine and trazodone inhibited melanogenesis in B16 cells by a dose-dependent
manner at the non-cytotoxic concentrations. Based on the results of the present study,
seeking new melanogenesis inhibitors from generic drugs is an alternative approach to
developing new depigmenting agents in cosmeceuticals. Moreover, cinnarizine and
trazodone were proven to be good candidates as skin-whitening agents for treatment of
skin hyperpigmentation |
| 關鍵字： | cinnarizine; inhibition; melanogenesis; trazodone; tyrosinase |
| 著作名稱： | Chang TS*, Lin MY, Lin HJ (2010) Identifying 8-Hydroxynaringenin as a Suicide Substrate of Mushroom Tyrosinase. J. Cosmet. Sci. 61, 205-210. |
| 年度： | 2010 |
| 類別： |
期刊論文
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| 摘要： | A biotransformed metabolite of naringenin was isolated from the fermentation broth of Aspergillus oryzae fed with naringenin and identified as 8-hydroxynaringenin based on the mass, 1H-, and 13C-NMR spectral data. The compound showed characteristics of both an irreversible inhibitor and a substrate of mushroom tyrosinase in the preincubation and HPLC analysis. These results demonstrate that 8-hydroxynaringenin belongs to a suicide substrate of mushroom tyrosinase. The partition ratio between the compounds molecules in the formation of product and in the inactivation of the enzyme was determined to be 283 ± 21. The present studys results, together with our previous findings, which proved that both 8-hydroxydaidzein and 8-hydroxygenistein are suicide substrates of mushroom tyrosinase, show that 7,8,4-trihydroxyl functional groups on flavonoids skeleton play important roles in producing suicide substrate properties toward mushroom tyrosinase. |
| 關鍵字： | Aspergillus oryzae, 8-hydroxynaringenin, suicide substrate, tyrosinase |
| 著作名稱： | Chang TS*, Lin JJ (2010) Inhibitory Effect of Danazol on Melanogenesis in Mouse B16 Melanoma Cells. Arch. Pharm. Res.33: 1959-1965 [SCI Ranking 78 % (185/236) in the field of Pharm. Pharm., IF 1.159 in 2009]. |
| 年度： | 2010 |
| 類別： |
期刊論文
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| 摘要： | In the present study, we screened more than 200 generic drugs to verify their applicability as skin-lightening agents by using mouse B16 melanoma cells. Of these, danazol was found to inhibit melanogenesis in B16 cells in a dose-dependent manner with an IC50 value of 9.3 M. In addition, danazol reduced cellular tyrosinase activity in B16 cells but not directly inhibited murine tyrosinase activity in the cell-free system. Furthermore, western blotting analysis confirmed that danazol downregulated the level of tyrosinase protein in B16 cells, while reverse-transcription polymerase chain reaction (RT-PCR) analysis found that danazol did not downregulate the level of tyrosinase mRNA in the cells. These results demonstrated that danazol inhibits melanogenesis in B16 cells via reducing tyrosinase activity with a post-transcriptional regulation. |
| 關鍵字： | |
| 著作名稱： | Ding HY, Lin HC, Chang TS*(2009) Tyrosinase inhibitors isolated from the roots of Paeonia suffruticosa. J. Cosmet. Sci. 60, 347-352 [SCI Ranking 88% (55/62) in the field of App. Chem., IF 0.283 in 2007] . |
| 年度： | 2009 |
| 類別： |
期刊論文
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| 摘要： | The inhibition of mushroom tyrosinase by Paeonia suffruticosa root-derived materials was evaluated. Six tyrosinase inhibitors were isolated by ethanol extraction, n-hexane, ethyl acetate, n-BuOH, and water partition, silica gel column chromatography, Sephadex LH-20, Lobar PR-8, and high-performance liquid chromatography methods and identified as kaempferol (I), quercetin (II), mudanpioside B (III), benzoyloxypaeoniflorin (IV), mudanpioside H (V), and pentagalloyl--D-glucose (VI) on the basis of spectroscopic evidence. The inhibitory activities of the compound I to VI against mushroom tyrosinase were determined with IC50 values of 0.120, 0.108, 0.368, 0.453, 0.324, and 0.063, mM, respectively. The kinetic study indicated that all purified inhibitors acted competitively for the L-dopa binding site of the enzyme with an exception of compound VI which acted non-competitively.
|
| 關鍵字： | |
| 著作名稱： | Chang TS*(2009) 8-Hydroxydaidzein is unstable in alkaline solutions. J. Cosmet. Sci. 60, 353-357 [ SCI Ranking 88% (55/62) in the field of App. Chem., IF 0.283 in 2007]. |
| 年度： | 2009 |
| 類別： |
期刊論文
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| 摘要： | 8-Hydroxydaidzein is a suicide substrate of mushroom tyrosinase with potent irreversible inhibitory activity. Despite its highly potential in the cosmetics industry, it was found that 8-hydroxydaidzein was unstable in the formulated cream. In this technical note, stabilities of 8-hydroxydaidzein in various solutions were investigated. The compound was dissolved in a series of solvents and the residual 8-hydroxydaidzeins in the prepared solution were sequentially determined during storage by HPLC. As a result, the loss in time of 8-hydroxydaidzein in both pH 6.8 phosphate buffer and DMSO showed typical first-order kinetics, and the loss rate constant of the compound in pH 6.8 phosphate buffer (4.48 x 10-3 hour-1) was 18-fold higher than that in DMSO (2.5 x 10-4 hour-1). The stabilities of the compound in different buffers with pH values ranging from pH 5 to pH 9 were determined in advance. The results showed that the compound was completely degraded in one day in the pH 8 and pH 9 buffers. In contrast, 8-hydroxydaidzein remained above 85% after 20 days’ storage in the pH 5 and pH 6 buffers. In addition to the residual 8-hydroxydaidzein analysis, the residual bioactivities including tyrosinase inhibitory activity and DPPH-radical scavenging activity of the 8-hydroxydaidzein solutions after 20 days’ storage in different pH values were also determined, and the results correlated well with that of the stability experiments. All the results demonstrated that 8-hydroxydaidzein is unstable in alkaline solutions. According to the data in the present report, it is recommended that 8-hydroxydaidzein should be formulated in an acid solution for its applications.
|
| 關鍵字： | Key words: |
| 著作名稱： | Chang TS*(2009) An updated review on tyrosinase inhibitors. Int.J. Mol. Sci. 10, 2440-2475 [Invited review, SCI Ranking 64% (81/127) in the field of Multidisciplinary Chem., IF 0.75 in 2007]
|
| 年度： | 2009 |
| 類別： |
期刊論文
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| 摘要： | Tyrosinase is a multifunctional, glycosylated, and copper-containing oxidase, which catalyzes the first step in mammalian melanogenesis and is responsible for enzymatic browning reactions in damaged fruits during post-harvest handling and processing. Neither hyperpigmentation in human skin nor enzymatic browning in fruits are desirable. These phenomena have encouraged researchers to seek new potent tyrosinase inhibitors for use in foods and cosmetics. This article surveys tyrosinase inhibitors newly discovered from natural and synthetic sources. The inhibitory strength is compared with that of a standard inhibitor, kojic acid, and their inhibitory mechanisms are discussed.
|
| 關鍵字： | Keywords: Browning; inhibitors; melanogenesis; tyrosinase. |
| 著作名稱： | Tai SS, Lin CG, Wu MH, Chang TS* (2009) Evaluation of Depigmenting Activity by 8-Hydroxydaidzein in Mouse B16 Melanoma Cells and Human Volunteers. Int. J. Mol. Sci. 10, 4257-4266 [SCI Ranking 57% (72/125) in the field of Multidisciplinary Chem., IF 0.978 in 2008 |
| 年度： | 2009 |
| 類別： |
期刊論文
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| 摘要： | In our previous study, 8-hydroxydaidzein (8-OHDe) was demonstrated to be a potent and unique suicide substrate of mushroom tyrosinase. In this study, the compound was evaluated for in vitro cellular tyrosinase and melanogenesis inhibitory activities in mouse B16 melanoma cells and for in vivo skin-whitening activity in human volunteers. Tyrosinase activity and melanogenesis in the cell culture incubated with 10 uM of 8-OHDe were decreased to 20.1% and 51.8% of control, respectively, while no obvious cytotoxicity was observed in this concentration. In contrast, a standard tyrosinase inhibitor, kojic acid, showed 69.9% and 71.3% of control in cellular tyrosinase and melanogenesis activity, respectively, at a concentration as high as 100 uM. Hence, 8-OHDe exhibited more than an inhibitory effects on melanin production in B16 cells 10-fold stronger than kojic acid. In addition, when a cream containing 4% 8-OHDe was applied to human skin in an in vivo study, significant increases in the dL*-values were observed after three weeks. Moreover, the increase in the dL*-values after 8-week treatment with 4% 8-OHDe (from -0.57 to 1.94) is stronger than those of 2% 8-OHDe treatment (from 0.26 to 0.94) and 2% ascorbic acid-2-glucoside treatment (from 0.07 to 1.54). From the results of the study, it was concluded that 8-OHDe, the potent suicide substrate of mushroom tyrosinase, has depigmenting activities in both mouse melanoma cells and in human volunteers. Thus, the compound has significant potential for use in cosmetics as a skin-whitening ingredient.
|
| 關鍵字： | Keywords: 8-Hydroxydaidzein; suicide substrate; skin whitening; tyrosinase inhibitor |
| 著作名稱： | Chang TS*, Tseng M, Ding HY, Tai SSK(2008)Isolation and Characterization of Streptomyces hiroshimensis strain TI-C3 with anti-tyrosinase activity. J Cosmet Sci (Accepted, SCI)
|
| 年度： | 2008 |
| 類別： |
期刊論文
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| 摘要： | |
| 關鍵字： | |
| 著作名稱： | Chang TS* (2007) Two potent suicide substrates of mushroom tyrosinase: 7,8,4’-trihydroxyisoflavone and 5,7,8,4’-tetrahydroxyisoflavone. J Agric Food Chem 55: 2010 – 2015 [SCI Ranking 8% (8/96) in the field of Food Science & Technology, IF 2.322 in 2006]. |
| 年度： | 2007 |
| 類別： |
期刊論文
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| 摘要： | Abstract-The inhibitory characteristics of two isoflavone metabolites, 7,8,4-trihydroxyisoflavone and 5,7,8,4-tetrahydroxyisoflavone, on mushroom tyrosinase were investigated. The two isoflavones were isolated from soygerm koji and inhibited both monophenolase and diphenolase activities of tyrosinase. Their inhibition type was demonstrated to be irreversible inhibition by pre-incubation and recovery experiments. By using HPLC analysis, it was found that mushroom tyrosinase could catalyze the two isoflavones. These results revealed that the two isoflavones belonged to suicide substrates of mushroom tyrosinase. The partition ratios between molecules of suicide substrate in the formation of product and in the inactivation of enzyme were determined to be 81.7 ± 5.9 and 35.5 ± 3.8, for 7,8,4-trihydroxyisoflavone and 5,7,8,4-tetrahydroxyisoflavone, respectively. From kinetic studies, maximal inactivation rate constants and Michaelis constants were 0.79 ± 0.08 and 1.01 ± 0.04 /min and 18.7 ± 2.31 and 7.81 ± 0.05 M for 7,8,4-trihydroxyisoflavone and 5,7,8,4-tetrahydroxyisoflavone, respectively, when the L-DOPA was used as the enzyme substrate. The structure analysis by comparing the inactivating activity between the two isoflavones and their structure analogs showed not only the 7,8-dihydroxyl groups but also the isoflavone skeleton of the two isoflavones played an important role in inactivating tyrosinase activity. The present study demonstrated that 7,8,4-trihydroxyisoflavone and 5,7,8,4-tetrahydroxyisoflavone are potent suicide substrates of mushroom tyrosinase.
|
| 關鍵字： | KEYWORDS: Inhibitor; irreversible; isoflavone; suicide substrate; tyrosinase |
| 著作名稱： | Chang TS*, Ding HY, Tai SSK, Wu CY, (2007) Metabolism of the soy isoflavone daidzein and genistein by the fungi used for the preparation of various fermented soybean foods. Biosci Biotechnol Biochem 71: 1330 - 1333 [SCI Ranking 30% (28/96) in the field of Food Science & Technology, IF 1.256 in 2006]. |
| 年度： | 2007 |
| 類別： |
期刊論文
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| 摘要： | The ability of fungi used in the preparation of fermented soybean foods to metabolize the soy isoflavones daidzein and genistein was investigated. A total of 21 fungal strains from dou-chi, miso, sake, soy sauce, and sufu were screened. The genera of the tested fungi included Actinomucor, Aspergillus, Candida, Debaryomyces, Monascus, Mucor, Rhizopus, Saccharomyces, and Zygosaccharomyces. The results were that all tested Aspergillus strains from these soybean foods, including five A. oryzae strains, one A. sojae strain, and one A. tamarii strain, metabolized both daidzein and genistein. In contrast, no other tested fungi from the fermented soybean foods metabolized either daidzein or genistein. The metabolites of daidzein and genistein by Aspergillus strains were identified as 8-hydroxydaidzein and 8-hydroxygenistein, respectively, based on their mass, 1H-, and 13C-NMR spectra.
|
| 關鍵字： | Key words: daidzein; fermented soybean foods; fungi; genistein; isoflavones |
| 著作名稱： | Chang TS*, Ding HY, Tai SSK, Wu CY,(2007)Tyrosinase Inhibitors Isolated from Soygerm Koji Fermented with Aspergillus oryzae BCRC 32288. Food Chem 105: 1430 – 1438 [SCI Ranking 6% (6/96) in the field of Food Science & Technology, IF 2.433 in 2006].
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| 年度： | 2007 |
| 類別： |
期刊論文
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| 摘要： | The inhibition of mushroom tyrosinase in soygerm koji fermented with Aspergillus oryzae BCRC 32288 was investigated. A methanol extract of the soygerm koji was partitioned into hexane, ethyl acetate and water. The ethyl acetate extract showed potent anti-tyrosinase activity with IC50 value of 0.19 mg/ml. The active compounds were isolated by activity-guided silica gel column chromatography and high-performance liquid chromatography (HPLC) method. Seven tyrosinase inhibitors were purified and identified as 6,7,4-trihydroxyisoflavone, 7,8,4-trihydroxyisoflavone, 5,7,8,4-tetrahydroxyisoflavone, 7,4-dihydroxyisoflavone (daidzein), 6-Methoxy-7,4-dihydroxyisoflavone (glycitein), 4-hydroxyisoflavone-7-O-glucoside (daidzin), and 5,4-dihydroxyisoflavone-7-O-glucoside (genistin) by comparing their mass, 1H-NMR, and 13C-NMR spectral data with those in the literatures. The purified seven isoflavones from fermented soygerm koji were divided into two groups based on their inhibitory characterizations on mushroom tyrosinase. Five isolated isoflavones showed inhibitory activity against monophenolase activity of mushroom tyrosinase only, with IC50 values of 0.009 ± 0.001 (6,7,4-trihydroxyisoflavone), 0.203 ± 0.018 (daidzein), 0.218 ± 0.007 (glycitein), 0.267 ± 0.008 (daidzin), and 0.343 ± 0.013 (genistin) mM. The kinetic study indicated that the five inhibitors significantly lengthened the lag time of the monophenolase activity of tyrosinase and acted competitively for the L-tyrosine binding site of the enzyme. So, the five isoflavones were competitive inhibitors on the monophenolase activity of tyrosinase. The other two isoflavones, 7,8,4-trihydroxyisoflavone and 5,7,8,4-tetrahydroxyisoflavone, inhibited both monophenolase and diphenolase activities of tyrosinase. Moreover, pre-incubation of each of the two isoflavones with tyrosinase resulted in total irreversible inhibition of the enzyme activity even under concentrations as low as of 10 M. Hence, 7,8,4-trihydroxyisoflavone and 5,7,8,4-tetrahydroxyisoflavone were irreversible inhibitors of mushroom tyrosinase.
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| 關鍵字： | Keywords: Inhibitor; Isoflavone; Irreversible; Soygerm; Tyrosinase |
| 著作名稱： | Chang TS* and Tseng M (2006) Preliminary screening of soil actinomycetes for anti-tyrosinase activity. J. Mar. Sci. Technol. 14, 190-193 [SCI Ranking 15% (1/7) in the field of Marine Engineering, IF 0.83 in 2008]. |
| 年度： | 2006 |
| 類別： |
期刊論文
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| 摘要： | |
| 關鍵字： | |
| 著作名稱： | Chang TS*, Ding HY, Lin HC (2005) Identifying 6,7,4’-Trihydroxyisoflavone as a Potent Tyrosinase Inhibitor. Biosci. Biotechnol. Biochem. 69, 1999-2001 [SCI Ranking 36% (38/107) in the field of Food Science & Technology, IF 1.39 in 2008]. |
| 年度： | 2005 |
| 類別： |
期刊論文
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| 摘要： | A known biotransformed compound, 6,7,4’-trihydroxyisoflavone, was identified as a potent tyrosinase inhibitor. It inhibited mushroom tyrosinase with an IC50 value of 9.2 M, which is six times the anti-tyrosinase activity of kojic acid (IC50 54.4 M). The inhibition kinetics, analyzed by Lineweaver-Burk plots, indicated 6,7,4’-trihydroxyisoflavone to be a competitive inhibitor of tyrosinase when L-tyrosine was used as a substrate. Its biosynthesis precursors and analogs, including glycitein, daidzein, and genistein, showed little anti-tyrosinase activity. The results suggest that hydroxyl groups at the C-6 and C-7 positions of the isoflavone skeleton might play an important role in the expression of tyrosinase inhibitory activity.
|
| 關鍵字： | Key words: inhibitor; isoflavone; kojic acid; 6,7,4’-trihydroxyisoflavone; tyrosinase |
| 著作名稱： | Novel Glycosylation by Amylosucrase to Produce Glycoside Anomers |
| 年度： | 2022 |
| 類別： |
期刊論文
Biology
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| 摘要： | Glycosylation occurring at either lipids, proteins, or sugars plays important roles in many biological systems. In nature, enzymatic glycosylation is the formation of a glycosidic bond between the anomeric carbon of the donor sugar and the functional group of the sugar acceptor. This study found novel glycoside anomers without an anomeric carbon linkage of the sugar donor. A glycoside hydrolase (GH) enzyme, amylosucrase from Deinococcus geothermalis (DgAS), was evaluated to glycosylate ganoderic acid F (GAF), a lanostane triterpenoid from medicinal fungus Ganoderma lucidum, at different pH levels. The results showed that GAF was glycosylated by DgAS at acidic conditions pH 5 and pH 6, whereas the activity dramatically decreased to be undetectable at pH 7 or pH 8. The biotransformation product was purified by preparative high-performance liquid chromatography and identified as unusual α-glucosyl-(2→26)-GAF and β-glucosyl-(2→26)-GAF anomers by mass and nucleic magnetic resonance (NMR) spectroscopy. We further used DgAS to catalyze another six triterpenoids. Under the acidic conditions, two of six compounds, ganoderic acid A (GAA) and ganoderic acid G (GAG), could be converted to α–glucosyl-(2→26)-GAA and β–glucosyl-(2→26)-GAA anomers and α-glucosyl-(2→26)-GAG and β-glucosyl-(2→26)-GAG anomers, respectively. The glycosylation of triterpenoid aglycones was first confirmed to be converted via a GH enzyme, DgAS. The novel enzymatic glycosylation-formed glycoside anomers opens a new bioreaction in the pharmaceutical industry and in the biotechnology sector. |
| 關鍵字： | amylosucrase; Deinococcus geothermalis; Ganoderma lucidum; glycosyl hydrolase; saponin; triterpenoid |
| 著作名稱： | A New Stilbene Glucoside from Biotransformation-Guided Purification of Chinese Herb Ha-Soo-Oh |
| 年度： | 2022 |
| 類別： |
期刊論文
plants
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| 摘要： | Ha-Soo-Oh is a traditional Chinese medicine prepared from the roots of Polygonum multiflorum Thunb. The herb extract has been widely used in Asian countries as a tonic agent and nutritional supplement for centuries. To identify new bioactive compounds in Chinese herbs, the biotransformation-guided purification (BGP) process was applied to Ha-Soo-Oh with Bacillus megaterium tyrosinase (BmTYR) as a biocatalyst. The result showed that a major biotransformed compound could be purified using the BGP process with preparative high-performance liquid chromatography (HPLC), and it was confirmed as a new compound, 2,3,5,3′,4′-pentahydroxystilbene-2-O-β-glucoside (PSG) following mass and nucleic magnetic resonance (NMR) spectral analyses. PSG was further confirmed as a biotransformation product from 2,3,5,4′-tetrahydroxystilbene-2-O-β-glucoside (TSG) by BmTYR. The new PSG exhibited 4.7-fold higher 1,1-diphenyl-2-picrylhydrazine (DPPH) free radical scavenging activity than that of TSG. The present study highlights the potential usage of BGP in herbs to discover new bioactive compounds in the future.
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| 關鍵字： | Polygonum multiflorum; Ha-Soo-Oh; tyrosinase; biotransformation; hydroxylation; melanogenesis; antioxidant |
| 著作名稱： | A Novel Soy Isoflavone Derivative, 3′-Hydroxyglycitin, with Potent Antioxidant and Anti-α-Glucosidase Activity |
| 年度： | 2022 |
| 類別： |
期刊論文
plants
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| 摘要： | This study demonstrated the enzymatic hydroxylation of glycitin to 3′-hydroxyglycitin, confirming the structure by mass and nucleic magnetic resonance spectral analyses. The bioactivity assays further revealed that the new compound possessed over 100-fold higher 1,1-diphenyl-2-picrylhydrazine free-radical scavenging activity than the original glycitin, although its half-time of stability was 22.3 min. Furthermore, the original glycitin lacked anti-α-glucosidase activity, whereas the low-toxic 3′-hydroxyglycitin displayed a 10-fold higher anti-α-glucosidase activity than acarbose, a standard clinical antidiabetic drug. The inhibition mode of 3′-hydroxyglycitin was noncompetitive, with a Ki value of 0.34 mM. These findings highlight the potential use of the new soy isoflavone 3′-hydroxyglycitin in biotechnology industries in the future. |
| 關鍵字： | soy isoflavone; tyrosinase; biotransformation; hydroxylation; antioxidant; glucosidase |
| 著作名稱： | Antioxidant and anti-α-glucosidase activities of biotransformable dragon’s blood via predicted data mining approach |
| 年度： | 2023 |
| 類別： |
期刊論文
Process Biochemistry
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| 摘要： | |
| 關鍵字： | |
| 著作名稱： | Novel Ganoderma Triterpenoid Saponins from the Biotransformation-guided Purification of a Commercial Ganoderma Extract |
| 年度： | 2023 |
| 類別： |
期刊論文
Journal of Biosciences and Biotechnology
|
| 摘要： | |
| 關鍵字： | |
| 著作名稱： | Enzymatic Synthesis of Novel and Highly Soluble Puerarin Glucoside by Deinococcus geothermalis Amylosucrase |
| 年度： | 2022 |
| 類別： |
期刊論文
Molecules
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| 摘要： | Puerarin (daidzein-8-C-glucoside) is an isoflavone isolated from several leguminous plants of the genus Pueraria. Puerarin possesses several pharmacological properties; however, the poor solubility of puerarin limits its applications. To resolve this poor solubility, Deinococcus geothermalis amylosucrase (DgAS) was used to modify puerarin into more soluble derivatives. The results showed that DgAS could biotransform puerarin into a novel compound: puerarin-4′-O-α-glucoside. The biotransformation reaction was manipulated at different temperatures, pH values, sucrose concentrations, reaction times, and enzyme concentrations. The results showed that the optimal reaction condition was biotransformed by 200 μg/mL DgAS with 20% (w/v) sucrose at pH 6 and incubated at 40 °C for 48 h, and the optimal production yield was 35.1%. Puerarin-4′-O-α-glucoside showed 129-fold higher solubility than that of puerarin and, thus, could be further applied for pharmacological use in the future. |
| 關鍵字： | amylosucrase; glycosylation; puerarin |
| 著作名稱： | Application of biotransformation-guided purification in Chinese medicine: An example to produce butin from licorice. |
| 年度： | 2022 |
| 類別： |
期刊論文
Catalysts
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| 摘要： | Abstract: Natural compounds are considered treasures in biotechnology; however, in the past, the process of discovering bioactive compounds is time consuming, and the purification and validation of the biofunctions and biochemistry of compounds isolated from a medicinal herb are tedious tasks. In this study, we developed an economical process called biotransformation‐guided purification (BGP), which we applied to analyze licorice, a traditional Chinese medicine widely used in many therapies. This medicinal herb contains various flavonoids and triterpenoids and, thus, is a suitable material used to assess the ability of BGP to identify and produce bioactive compounds. In the BGP process, the ethyl acetate extract of a commercial licorice medicine was partially purified into three fractions by Sephadex LH‐20 chromatography, and Bacillus megaterium tyrosinase (BmTYR) was used to catalyze the biotransformation of the extract from each fraction. One of the products pro‐ duced via BmTYR‐driven biotransformation was purified from the biotransformation‐positive ex‐ tract using preparative C‐18 high‐performance liquid chromatography, and it was identified as butin (3′‐hydroxyliquiritigenin) through nucleic magnetic resonance and mass spectral analyses. Butin was produced from liquiritigenin through BmTYR‐catalyzed hydroxylation, with commercial liquiritigenin as the biotransformation precursor. The proposed alternative approach quickly iden‐ tified and isolated the biotransformed butin from licorice. Moreover, butin demonstrated an anti‐ oxidant activity that is stronger by over 100‐fold compared with that of its precursor (liquiritigenin). This study showed that the economical BGP process could quickly obtain and validate bioactive molecules from crude extracts of medicinal herbs.
|
| 關鍵字： | Glycyrrhiza; licorice; tyrosinase; biotransformation; hydroxylation; antioxidant |
| 著作名稱： | Potential Industrial Production of a Well-Soluble, Alkaline-Stable, and Anti-Inflammatory Isoflavone Glucoside from 8-Hydroxydaidzein Glucosylated by Recombinant Amylosucrase of Deinococcus geothermalis |
| 年度： | 2019 |
| 類別： |
期刊論文
Moleculues
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| 摘要： | : 8-Hydroxydaidzein (8-OHDe), an ortho-hydroxylation derivative of soy isoflavone
daidzein isolated from some fermented soybean foods, has been demonstrated to possess potent
anti-inflammatory activity. However, the isoflavone aglycone is poorly soluble and unstable in
alkaline solutions. To improve the aqueous solubility and stability of the functional isoflavone,
8-OHDe was glucosylated with recombinant amylosucrase of Deinococcus geothermalis (DgAS) with
industrial sucrose, instead of expensive uridine diphosphate-glucose (UDP-glucose). One major
product was produced from the biotransformation, and identified as 8-OHDe-7-α-glucoside, based
on mass and nuclear magnetic resonance spectral analyses. The aqueous solubility and stability of
the isoflavone glucoside were determined, and the results showed that the isoflavone glucoside was
almost 4-fold more soluble and more than six-fold higher alkaline-stable than 8-OHDe. In addition,
the anti-inflammatory activity of 8-OHDe-7-α-glucoside was also determined by the inhibition
of lipopolysaccharide-induced nitric oxide production in RAW 264.7 cells. The results showed
that 8-OHDe-7-α-glucoside exhibited significant and dose-dependent inhibition on the production
of nitric oxide, with an IC50 value of 173.2 µM, which remained 20% of the anti-inflammatory
activity of 8-OHDe. In conclusion, the well-soluble and alkaline-stable 8-OHDe-7-α-glucoside
produced by recombinant DgAS with a cheap substrate, sucrose, as a sugar donor retains moderate
anti-inflammatory activity, and could be used in industrial applications in the future. |
| 關鍵字： | 8-hydroxydaidzein; stable; soluble; anti-inflammation; amylosucrase;Deinococcus geothermalis |
| 著作名稱： | A New Triterpenoid Glucoside from a Novel Acidic Glycosylation of Ganoderic Acid A via Recombinant Glycosyltransferase of Bacillus subtilis. |
| 年度： | 2019 |
| 類別： |
期刊論文
Molecules
|
| 摘要： | Ganoderic acid A (GAA) is a bioactive triterpenoid isolated from the medicinal fungus Ganoderma lucidum. Our previous study showed that the Bacillus subtilis ATCC (American type culture collection) 6633 strain could biotransform GAA into compound (1), GAA-15-O-β-glucoside, and compound (2). Even though we identified two glycosyltransferases (GT) to catalyze the synthesis of GAA-15-O-β-glucoside, the chemical structure of compound (2) and its corresponding enzyme remain elusive. In the present study, we identified BsGT110, a GT from the same B. subtilis strain, for the biotransformation of GAA into compound (2) through acidic glycosylation. BsGT110 showed an optimal glycosylation activity toward GAA at pH 6 but lost most of its activity at pH 8. Through a scaled-up production, compound (2) was successfully isolated using preparative high-performance liquid chromatography and identified to be a new triterpenoid glucoside (GAA-26-O-β-glucoside) by mass and nuclear magnetic resonance spectroscopy. The results of kinetic experiments showed that the turnover number (kcat) of BsGT110 toward GAA at pH 6 (kcat 11.2 min−1) was 3-fold higher than that at pH 7 (kcat 3.8 min−1), indicating that the glycosylation activity of BsGT110 toward GAA was more active at acidic pH 6. In short, we determined that BsGT110 is a unique GT that plays a role in the glycosylation of triterpenoid at the C-26 position under acidic conditions, but loses most of this activity under alkaline ones, suggesting that acidic solutions may enhance the catalytic activity of this and similar types of GTs toward triterpenoids. |
| 關鍵字： | ganoderic acid A; glucosyltransferase; acidic; Bacillus subtilis; triterpenoid |
| 著作名稱： | A Genome-Centric Approach Reveals a Novel Glycosyltransferase from the GA A07 Strain of Bacillus thuringiensis Responsible for Catalyzing 15-O-Glycosylation of Ganoderic Acid A. |
| 年度： | 2019 |
| 類別： |
期刊論文
Int. J. Mol. Sci.
|
| 摘要： | Abstract: Strain GA A07 was identified as an intestinal Bacillus bacterium of zebrafish, which has high efficiency to biotransform the triterpenoid, ganoderic acid A (GAA), into GAA-15-O-β-glucoside. To date, only two known enzymes (BsUGT398 and BsUGT489) of Bacillus subtilis ATCC 6633 strain can biotransform GAA. It is thus worthwhile to identify the responsible genes of strain GA A07 by whole genome sequencing. A complete genome of strain GA A07 was successfully assembled. A phylogenomic analysis revealed the species of the GA A07 strain to be Bacillus thuringiensis. Forty glycosyltransferase (GT) family genes were identified from the complete genome, among which three genes (FQZ25_16345, FQZ25_19840 , and FQZ25_19010) were closely related to BsUGT398 and BsUGT489. Two of the three candidate genes, FQZ25_16345 and FQZ25_19010, were successfully cloned and expressed in a soluble form in Escherichia coli, and the corresponding proteins, BtGT_16345 and BtGT_19010, were purified for a biotransformation activity assay. An ultra-performance liquid chromatographic analysis further confirmed that only the purified BtGT_16345 had the key biotransformation activity of catalyzing GAA into GAA-15-O-β-glucoside. The suitable conditions for this enzyme activity were pH 7.5, 10 mM of magnesium ions, and 30 °C. In addition, BtGT_16345 showed glycosylation activity toward seven flavonoids (apigenein, quercetein, naringenein, resveratrol, genistein, daidzein, and 8-hydroxydaidzein) and two triterpenoids (GAA and antcin K). A kinetic study showed that the catalytic efficiency (kcat/KM) of BtGT_16345 was not significantly different compared with either BsUGT398 or BsUGT489. In short, this study identified BtGT_16345 from B. thuringiensis GA A07 is the catalytic enzyme responsible for the 15-O-glycosylation of GAA and it was also regioselective toward triterpenoid substrates. |
| 關鍵字： | Keywords: nanopore sequencing; ganoderic acid; Bacillus thuringiensis; biotransformation; glycosyltransferase; whole genome sequencing |
| 著作名稱： | Sequential Biotransformation of Antcin K by Bacillus subtilis ATCC 6633. |
| 年度： | 2018 |
| 類別： |
期刊論文
Catalysts
|
| 摘要： | The biotransformation of antcin K, a major ergostane triterpenoid from the fruiting bodies of Antrodia cinnamomea, by Bacillus subtilis (B. subtilis) ATCC 6633 was studied. Four metabolites from the biotransformation were isolated with preparative high-performance liquid chromatography and identified as 25S-antcin K 26-O--glucoside, 25R-antcin K 26-O--glucoside, 25S-antcin K 26-O--(6’-O-succinyl)-glucoside, and 25R-antcin K 26-O--(6’-O-succinyl)-glucoside with mass and nuclear magnetic resonance spectral analysis. By using either 25S-antcin K 26-O--glucoside or 25R-antcin K 26-O--glucoside as the biotransformation precursor, it was proven that 25S-antcin K 26-O--(6’-O-succinyl)-glucoside and 25R-antcin K 26-O--(6’-O-succinyl)-glucoside were biotransformed from 25S-antcin K 26-O--glucoside and 25R-antcin K 26-O--glucoside, respectively. To the best of our knowledge, this is the first study on the glycosylation of triterpenoids from A. cinnamomea, and the first time the succinylation of triterpenoid glycosides by microorganisms has been found. In addition, all four antcin K glucoside derivatives are new compounds. |
| 關鍵字： | Biotransformation, Antcin K, Bacillus subtilis |
| 著作名稱： | Production of New Isoflavone Glucosides from Glycosylation of 8-Hydroxydaidzein by Glycosyltransferase from Bacillus subtilis ATCC 6633. |
| 年度： | 2018 |
| 類別： |
期刊論文
Catalysts
|
| 摘要： | 8-Hydroxydaidzein (8-OHDe) has been proven to possess some important bioactivities; however, the low aqueous solubility and stability of 8-OHDe limit its pharmaceutical and cosmeceutical applications. The present study focuses on glycosylation of 8-OHDe to improve its drawbacks in solubility and stability. According to the results of phylogenetic analysis with several identified flavonoid-catalyzing glycosyltransferases (GTs), three glycosyltransferase genes (BsGT110, BsGT292 and BsGT296) from the genome of the Bacillus subtilis ATCC 6633 strain were cloned and expressed in Escherichia coli. The three BsGTs were then purified and the glycosylation activity determined toward 8-OHDe. The results showed that only BsGT110 possesses glycosylation activity. The glycosylated metabolites were then isolated with preparative high-performance liquid chromatography and identified as two new isoflavone glucosides, 8-OHDe-7-O-β-glucoside and
8-OHDe-8-O-β-glucoside, whose identity was confirmed by mass spectrometry and nuclear magnetic resonance spectroscopy. The aqueous solubility of 8-OHDe-7-O-β-glucoside and
8-OHDe-8-O-β-glucoside is 9.0- and 4.9-fold, respectively, higher than that of 8-OHDe. Moreover, more than 90% of the initial concentration of the two 8-OHDe glucoside derivatives remained after 96 h of incubation in 50 mM of Tris buffer at pH 8.0. In contrast, the concentration of 8-OHDe decreased to 0.8% of the initial concentration after 96 h of incubation. The two new isoflavone glucosides might have potential in pharmaceutical and cosmeceutical applications.
|
| 關鍵字： | 8-Hydroxydaidzein, Glycosyltransferase, Bacillus |
| 著作名稱： | New Triterpenoid from Novel Triterpenoid 15-O-Glycosylation on Ganoderic Acid A by Intestinal Bacteria of Zebrafish. |
| 年度： | 2018 |
| 類別： |
期刊論文
Molecules
|
| 摘要： | Functional bacteria that could biotransform triterpenoids may exist in the diverse microflora of fish intestines. Ganoderic acid A (GAA) is a major triterpenoid from the medicinal fungus Ganoderma lucidum. In studying the microbial biotransformation of GAA, dozens of intestinal bacteria were isolated from the excreta of zebrafish. The bacteria’s ability to catalyze GAA were determined using ultra-performance liquid chromatography analysis. One positive strain, GA A07, was selected for functional studies. GA A07 was confirmed as Bacillus sp., based on the DNA sequences of the 16S rRNA gene. The biotransformed metabolite was purified with the preparative high-performance liquid chromatography method and identified as GAA-15-O-β-glucoside, based on the mass and nuclear magnetic resonance spectral data. The present study is the first to report the glycosylation of Ganoderma triterpenoids. Moreover, 15-O-glycosylation is a new microbial biotransformation of triterpenoids, and the biotransformed metabolite, GAA-15-O-β-glucoside, is a new compound. |
| 關鍵字： | Triterpenoid , Ganoderic Acid A, Intestinal Bacteria |
| 著作名稱： | Uridine Diphosphate-Dependent Glycosyltransferases from Bacillus subtilis ATCC 6633 Catalyze the 15-O-Glycosylation of Ganoderic Acid A. |
| 年度： | 2018 |
| 類別： |
期刊論文
Int. J. Mol. Sci.
|
| 摘要： | Bacillus subtilis ATCC 6633 was found to biotransform ganoderic acid A (GAA), which is a major lanostane triterpenoid from the medicinal fungus Ganoderma lucidum. Five glycosyltransferase family 1 (GT1) genes of this bacterium, including two uridine diphosphate-dependent glycosyltransferase (UGT) genes, BsUGT398 and BsUGT489, were cloned and overexpressed in Escherichia coli. Ultra-performance liquid chromatography confirmed the two purified UGT proteins biotransform ganoderic acid A into a metabolite, while the other three purified GT1 proteins cannot biotransform GAA. The optimal enzyme activities of BsUGT398 and BsUGT489 were at pH 8.0 with 10 mM of magnesium or calcium ion. In addition, no candidates showed biotransformation activity toward antcin K, which is a major ergostane triterpenoid from the fruiting bodies of Antrodia cinnamomea. One biotransformed metabolite from each BsUGT enzyme was then isolated with preparative high-performance liquid chromatography. The isolated metabolite from each BsUGT was identified as ganoderic acid A-15-O-β-glucoside by mass and nuclear magnetic resonance spectroscopy. The two BsUGTs in the present study are the first identified enzymes that catalyze the 15-O-glycosylation of triterpenoids. |
| 關鍵字： | Glycosyltransferases, Bacillus subtilis, Ganoderic Acid A |
| 著作名稱： | Biotransformation of Ganoderic Acid A to 3-O-Acetyl Ganoderic Acid A by Soil-isolated Streptomyces sp. |
| 年度： | 2018 |
| 類別： |
期刊論文
Fermentation
|
| 摘要： | The medicinal fungus Ganoderma lucidum contains many bioactive triterpenoids, ganoderic acid A (GAA) being one of the major ones. The present study explored the microbial biotransformation of GAA, isolating 283 strains of soil actinomycetes and determining their abilities to biotransform GAA with ultra-performance liquid chromatography analysis. One positive strain, AI 045, was selected to validate the biotransformation activity. The strain was identified as Streptomyces sp. based on the sequenced 16S rRNA gene. The produced compound obtained from the biotransformation of GAA was purified with the preparative high-performance liquid chromatography method and identified as 3-O-acetyl GAA based on mass and nuclear magnetic resonance spectral data. The present study is the first report that bacteria have the novel ability to biotransform the triterpenoids of fungus G. lucidum. Moreover, the identified 3-O-acetyl GAA is a new triterpenoid product discovered in microbes. |
| 關鍵字： | Biotransformation, Ganoderic Acid A, Streptomyces |
| 著作名稱： | Production and anti-melanoma activity of methoxyisoflavones from biotransformation of genistein by two recombinant Escherichia coli strains. |
| 年度： | 2017 |
| 類別： |
期刊論文
Molecules
|
| 摘要： | Biotransformation of the soy isoflavone genistein by sequential 30-hydroxylation using
recombinant Escherichia coli expressing tyrosinase from Bacillus megaterium and then methylation using another recombinant E. coli expressing O-methyltransferase from Streptomyces peucetius was conducted. The results showed that two metabolites were produced from the biotransformation, identified as 5,7,40-trihydroxy-30-methoxyisoflavone and 5,7,30-trihydroxy-40-methoxyisoflavone, respectively, based on their mass and nuclear magnetic resonance spectral data. 5,7,40-Trihydroxy-30-methoxyisoflavone showed potent antiproliferative activity toward mouse B16 melanoma cells with an IC50 value of 68.8 M. In contrast, the compound did not show any cytotoxicity towardmouse normal fibroblast cells, even at 350 M concentration. The results of the present study offer insight on the production of both 5,7,40-trihydroxy-30-methoxyisoflavone and 5,7,30-trihydroxy-40-methoxyisoflavone by two recombinant E. coli strains and the potential anti-melanoma applications of 5,7,40-trihydroxy-30-methoxyisoflavone. |
| 關鍵字： | |
| 著作名稱： | Biotransformation of Ergostane Triterpenoid Antcin K from Antrodia cinnamomea by Soil-Isolated Psychrobacillus sp. AK 1817 |
| 年度： | 2017 |
| 類別： |
期刊論文
Catalysts
|
| 摘要： | Antcin K is one of the major ergostane triterpenoids from the fruiting bodies of Antrodia cinnamomea, a parasitic fungus that grows only on the inner heartwood wall of the aromatic tree Cinnamomum kanehirai Hay (Lauraceae). To search for strains that have the ability to biotransform antcin K, a total of 4311 strains of soil bacteria were isolated, and their abilities to catalyze antcin K were determined by ultra-performance liquid chromatography analysis. One positive strain, AK 1817, was selected for functional studies. The strain was identified as Psychrobacillus sp., based on the DNA sequences of the 16S rRNA gene. The biotransformation metabolites were purified with the preparative high-performance liquid chromatography method and identified as antcamphin E and antcamphin F, respectively, based on the mass and nuclear magnetic resonance spectral data. The present study is the first to report the biotransformation of triterpenoids from A. cinnamomea (Antrodia cinnamomea). |
| 關鍵字： | Antrodia cinnamomea; biotransformation; antcin K; triterpenoid; Psychrobacillus |
| 著作名稱： | Improving free radical scavenging activity of soy isoflavone glycosides daidzin and genistin by 3’-hydroxylation using recombinant Escherichia coli. |
| 年度： | 2016 |
| 類別： |
期刊論文
Molecules
|
| 摘要： | The present study describes the biotransformation of a commercially available crude extract
of soy isoflavones, which contained significant amounts of the soy isoflavone glycosides daidzin
and genistin, by recombinant Escherichia coli expressing tyrosinase from Bacillus megaterium. Two major products were isolated from the biotransformation and identified as 30-hydroxydaidzin and 30-hydroxygenistin, respectively, based on their mass and nuclear magnetic resonance spectral data. The two 30-hydroxyisoflavone glycosides showed potent 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity with IC50 values of 7.4 and 9.8 M for 30-hydroxydaidzin and 30-hydroxygenistin, respectively. The free radical scavenging activities of the two 30-hydroxyisoflavone glycosides were, respectively, 120 and 72 times higher than the activity of their precursors, daidzin and genistin, and were also stronger than the activity of ascorbic acid, which showed an IC50 value of 15.1 M. This is the first report of the bio-production and potential antioxidant applications of both 30-hydroxydaidzin and 30-hydroxygenistin. |
| 關鍵字： | antioxidant; biotransformation; daidzin; genistin; hydroxylation |
| 著作名稱： | Improving 3-hydroxygenistein production in recombinant Pichia pastoris by a periodic hydrogen peroxide-shocking strategy |
| 年度： | 2016 |
| 類別： |
期刊論文
Journal of Microbiology and Biotechnolgoy
|
| 摘要： | 3-Hydroxygenistein can be obtained from the biotransformation of genistein by the engineered Pichia pastoris X-33 strain, which harbors a fusion gene composed of CYP57B3 from Aspergillus oryzae and a cytochrome P450 oxidoreductase gene (sCPR) from Saccharomyces cerevisiae. P. pastoris X-33 mutants with higher 3-hydroxygenistein production were selected through a periodic hydrogen peroxide-shocking strategy. One mutant (P2-D14-5) was found to produce 23.0 mg/l of 3-hydroxygenistein, which is an amount 1.87-fold higher than that produced by the recombinant X-33. In a 5-L fermenter, 20.3 mg/l of 3-hydroxygenistein was produced by P2-D14-5. The P2-D14-5 mutant therefore has high potential for industrial-scale 3-hydroxygenistein production. |
| 關鍵字： | 3-Hydroxygenistein, orobol, CYP57B3, Pichia pastoris |
| 著作名稱： | Production of Two Novel Methoxy-Isoflavones from
Biotransformation of 8-Hydroxydaidzein by
Recombinant Escherichia coli Expressing
O-Methyltransferase SpOMT2884 from
Streptomyces peucetius |
| 年度： | 2015 |
| 類別： |
期刊論文
International Journal of Molecular Science
|
| 摘要： | Biotransformation of 8-hydroxydaidzein by recombinant Escherichia coli expressing
O-methyltransferase (OMT) SpOMT2884 from Streptomyces peucetius was investigated. Two
metabolites were isolated and identified as 7,41-dihydroxy-8-methoxy-isoflavone (1) and
8,41-dihydroxy-7-methoxy-isoflavone (2), based on mass, 1H-nuclear magnetic resonance (NMR)
and 13C-NMR spectrophotometric analysis. The maximum production yields of compound (1) and
(2) in a 5-L fermenter were 9.3 mg/L and 6.0 mg/L, respectively. The two methoxy-isoflavones
showed dose-dependent inhibitory effects on melanogenesis in cultured B16 melanoma cells under
non-toxic conditions. Among the effects, compound (1) decreased melanogenesis to 63.5% of the
control at 25 M. This is the first report on the 8-O-methylation activity of OMT toward isoflavones.
In addition, the present study also first identified compound (1) with potent melanogenesis
inhibitory activity. |
| 關鍵字： | 7,41-dihydroxy-8-methoxy-isoflavone; 8,41-dihydroxy-7-methoxy-isoflavone; 8-hydroxydaidzein; melanogenesis; inhibition; O-methyltransferase; SpOMT2884 |
| 著作名稱： | Methanol Partition Fraction of Ethanol of Discorea nipponica Makino Inhibits Melanogenesis |
| 年度： | 2014 |
| 類別： |
期刊論文
Tropical Journal of Pharmaceutical Research
|
| 摘要： | Abstract
Purpose: To investigate the inhibitory effects of methanol layer of Dioscorea nipponica Makino ethanolic extract (DNM) on melanogenesis both in vitro and in vivo.
Methods: Both in vitro cultured mouse B16 melanoma cell and in vivo zebrafish were used to evaluate the melanogenesis inhibitory activity of DNM. In B16 cells, inhibitory effects on intracellular melanogenesis, tyrosinase activity and reactive oxygen species (ROS) were determined after DNM treatment. In zebrafish, both toxic and antimelanogenic activities of DNM on developed larvae were evaluated.
Results: In B16 cells, results showed that DNM dose-dependently inhibited melanogenesis under nontoxic concentrations. Surprisingly, however, DNM didn’t reduce either intracellular tyrosinase activity or the protein amount of the enzyme in B16 cells. On the other hand, DNM showed strong antioxidant activities against cell-free 2,2-diphenyl-1-picryl-hydrazl (DPPH) and 2,2’-azino-bis (3-etnylbenzthiazoline-6-sulphaonic acid) (ABTS+) free radical and intracellular ROS in B16 cells. In zebrafish, DNM significantly and dose-dependently inhibited skin melanogenesis of zebrafish larvae under nontoxic concentrations.
Conclusions: The present study demonstrated that DNM inhibited melanogenesis in vitro in B16 melanoma cells and in vivo in zebrafish. Moreover, DNM exhibited the potent inhibition on melanogenesis not due to down-regulating intracellular tyrosinase activity, but possibly through antioxidant activity in the cells.
|
| 關鍵字： | Keywords: Dioscorea nipponica M., Melanogenesis, Tyrosinase, Antioxidant |
| 著作名稱： | Finding New Compounds via a Predicted Data Mining Approach |
| 年度： | 2023 |
| 類別： |
會議論文
|
| 摘要： | |
| 關鍵字： | |
| 著作名稱： | Inhibition of alpha-glucosidase by loureirin A and loureirin B |
| 年度： | 2023 |
| 類別： |
會議論文
|
| 摘要： | |
| 關鍵字： | |
| 著作名稱： | Using Copper-Containing Tyrosinase to Produce 3’-Hydroxyglycitein |
| 年度： | 2022 |
| 類別： |
會議論文
|
| 摘要： | |
| 關鍵字： | |
| 著作名稱： | Transglycosylation Activity of Bifidobacterium dentium Amylosucrase |
| 年度： | 2022 |
| 類別： |
會議論文
|
| 摘要： | |
| 關鍵字： | |
| 著作名稱： | Bacterial glycosyltransferase from Bacillus strains acting on Gaonderma triterpenoid |
| 年度： | 2021 |
| 類別： |
會議論文
|
| 摘要： | |
| 關鍵字： | |
| 著作名稱： | Inhibitory Effects of 8-Methoxydaidzein on Tyrosinase Activity in Mouse B16 Melanoma Cells |
| 年度： | 2017 |
| 類別： |
會議論文
|
| 摘要： | Our previous study has |
| 關鍵字： | Daidzein, 8-hydroxydaidzein, methylation, methyltransferase, melanogenesis, inhibition, tyrosinase |
| 著作名稱： | Biotransformation of Quercetin by Recombinant Escherichia coli Expressing O-Methyltransferase from Streptomyces peucetius |
| 年度： | 2016 |
| 類別： |
會議論文
|
| 摘要： | Biotransformation of Quercetin by Recombinant Escherichia coli Expressing O-Methyltransferase from Streptomyces peucetius
Chien-Min Chianga, Chun-Ping Jenb Te-Sheng Changc
a Department of Biotechnology, Chia Nan University of Pharmacy, No. 60, Sec. 1, Erh-Jen Rd., Jen-Te District, Tainan, Taiwan, R.O.C.
E-mail address: cmchiang@mail.cnu.edu.tw
b Department of Mechanical Engineering, National Chung Cheng University, Taiwan, R.O.C.
c Department of Biological Sciences and Technology, National University of Tainan, No. 33, Sec. 2, Shu-Lin St. Tainan, Taiwan, R.O.C.
E-mail address: mozyme2001@gmail.com
1. Background
O-Methylation modification is a part of biosynthesis of some isoflavones and plays a key role in the secondary metabolism in plants [1]. Enzymatic O-methylation is carried out by O-methyltransferase (OMT), which uses S-adenosylmethionine (SAM) as a methyl group donor. Our previous study demonstrated that recombinant Escherichia coli expressing O-methyltransferase from Streptomyces peucetius catalyzed both C-7 and C-8 methylation of 8-hydroxydaidzein (7,8,4-trihydroxyisoflavone) [2]. In the present study, the biotransformation of quercetin (2,5,7,3,4-tetrahydroxyflavone)by the recombinant E. coli was investigated.
2. Materials and Methods
Recombinant E. coli BL21 (DE3) harboring expression vector pETDuet-SpOMT was obtained from our previous study [2]. Quercetin and isopropyl--D-thiogalactopyranoside (IPTG) were purchased from Sigma (St. Louis, MO). The recombinant E. coli harboring the expression vector were cultivated in 20 mL of LeMaster and Richards minimal medium (LR medium) containing 50 g/ml of ampicillin and 0.4% glycerol, with 200 rpm shaking at 37oC. As the optical density at 600 nm reached 0.6, 0.5 mM of IPTG and 0.1 mM of quercetin were added to induce expression of the OMT gene and start biotransformation. At indicated time intervals, aliquots of cells were harvested, extracted with MeOH/ACN (50%:50%), and analyzed by ultra performance liquid chromatography (UPLC). The operational conditions of UPLC were the same as our previous work [2].
3. Results and Discussion
For studying biotransformation, the substrate quercetin was added together with IPTG in E. coli cultivation, and the products profile were determined by UPLC to check whether the substrate could be converted by the recombinant cells. As shown in Figure 1A, one major metabolite appeared as a new peak at retention time of 5.0 min in the profile of fermentation broth at 24 h of incubation. The result proved that the recombinant E. coli also catalyzed quercetin. Our previous study showed that the recombinant E. coli catalyzed both C-8 and C-7 methylation toward 8-hydroxydaidzin. However, the present study showed that only one metabolite appeared during the biotransformation and implied that the recombinant E. coli catalyzed methylation at single carbon position of quercetin. The purification and then resolving the chemical structure of the metabolite is in progress.
The production profile of the metabolite is shown in Figure 1B. In the result, the product was accumulated rapidly within the initial 6 h after induction and then followed with a very slow rate. The possible reason for the decreased enzymatic conversion rate after 6 h of induction is insufficiency of cofactor SAM. We tried to feed SAM into the cultivation at 6 and/or 18 h of induction and did not increase the production (data not shown). It has been reported that co-expression of SAM synthetase gene could increase OMT activity. Further study should be conducted to evaluate the effect of co-expression of SAM synthetase gene on the biotransformation of quercetin by the recombinant E. coli.
Keywords: Quercetin, O-Methyltransferase, Methylation, Streptomyces peucetius
References
[1] Kim BG, Sung SH, Chong SY, Lim Y, Ahn JH. Plant flavonoid O-methyltransferase: substrate specificity and application. J Plant Biol 2010; 53: 321–329.
[2] Chiang CM, Ding HY, Ts |
| 關鍵字： | Quercetin, O-Methyltransferase, Methylation, Streptomyces peucetius |
| 著作名稱： | Biotransformation of 3’-Hydroxydaidzein by Escherichia coli Expressing O-Methyltransferase |
| 年度： | 2016 |
| 類別： |
會議論文
|
| 摘要： |
Two metabolites were produced from biotransformation reaction from 3’-hydroxydaidzein by recombinant Escherichia coli, which expressed O-methyltransferase (OMT) SpOMT2884 from Streptomyces peucetius. The two methoxy-isoflavones metabolites were isolated by using preparative high-pressure liquid chromatography (HPLC). Anti-melanogenesis activity of both methoxy-isoflavones metabolites were determined using cultured B16 melanoma cells. The results showed that one (metabolite 2) of the two methoxy-isoflavones metabolites dose-dependently inhibited melanogenesis under non-toxic conditions. Using NMR and Mass spectral analysis to resolve the chemical structure of the active compound was undertaken in our laboratory. The present study highlights the production and application of methoxy-isoflavones in the cosmetics industry.
|
| 關鍵字： | 3’-Hydroxydaidzein, Methyltransferase, Biotransformation |
| 著作名稱： | Improving substrate spectrum and product yields of CYP105D7 from Streptomyces avermitilis MA4680 through membrane-anchoring and reductase fusion expression in Pichia pastoris |
| 年度： | 2015 |
| 類別： |
會議論文
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| 摘要： | Biotransformation of soy isoflavones into ortho-hydroxyisoflavones by CYP105D7 from Streptomyces avermitilis MA4680 was investigated through chimeric expression in Pichia pastoris. Using N-terminal fusion with the transmembrane domain of CYP57B3 from Aspergillus oryzae and C-terminal fusion with a P450 reductase from Saccharomyces cerevisiae, CYP105D7 was expressed as a reductase fusion and membrane-anchoring form in P. pastoris. Recombinant P. pastoris expressing the chimera catalyzed both biotransformation of daidzein and genistein. This is the first study to show the catalyzing activity of CYP105D7 toward genistein. The major product from daidzein was identified as 6-hydroxydaidzein via comparing ultra-performance liquid chromatography analysis with the authentic standard. The major product from genistein was purified using preparative high-performance liquid chromatography and identified as 3-hydroxygenistein on the basis of nuclear magnetic resonance and mass data. The recombinant P. pastoris produced 6-hydroxydaidzein and 3-hydroxygenistein in a 5-L fermenter, with maximal yields of 7.5 and 15.0 mg/l, respectively, and the production of 3-hydroxygenistein was higher than any previously reported in the literature. The present study demonstrated that substrate spectrum and product yields of biotransformation from soy isoflavones into ortho-hydroxyisoflavones by bacterial CYP105D7 could be improved through membrane-anchoring and reductase fusion expression in P. pastoris.
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| 關鍵字： | CYP105D7, isoflavones, daidzein, genistein, Streptomyces |
| 著作名稱： | Production of 6-Hydroxyapigenin by Recombinant Pichia pastoris Harboring Fusion P450 Gene |
| 年度： | 2014 |
| 類別： |
會議論文
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| 摘要： | Biotransformation of apigenin by recombinant Pichia pastoris was investigated. The recombinant yeast harbored a fusion gene composed of CYP57B3 gene from Aspergillus oryzae and a cytochrome reductase gene from Saccharomyces cerevisiae. The recombinant P. pastoris was found to biotransform apigenin into 6-hydroxyapigenin (scutellarein). This is the first report about 6-hydroxylation of apigenin via microbial biotransformation.
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| 關鍵字： | 6-Hydroxyapigenin, Pichia, P450 |
| 著作名稱： | TRANSFORMATION OF DAIDZEIN BY A RECOMBINANT STRAIN OF PICHIA PASTORIS |
| 年度： | 2013 |
| 類別： |
會議論文
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| 摘要： | Aspergillus oryzae is the most well-known microorganism that transforms soyisoflavones into the corresponding ortho-hydroxyl derivatives. A P450 enzyme from the microorganism, CYP57B3, was recently shown to catalyze ortho-hydroxylation of soyisoflavone genistein by cooperating with a P450 reductase (CPR) from Saccharomyces cerevisiae (Nazir et al., 2011).Because of the skin-whitening activity of ortho-hydroxydaidzein derivatives (Chang et al., 2005; Chang et al., 2007; Chang, 2007; Tai et al., 2009), we investigate the biotransformation of daidzein by the CYP57B3 in the present study. We used an EazySelectTM Pichia expression system (Invitrogen) to study. In order to solve the requirement for redox proteins, which is considered to be a bottle neck in P450 enzyme reaction, we fused CYP57B3 with CPR from S. cerevisiae. Fig. 1 shows a diagram of the biotransformation of daidzein to ortho-hydroxydaidzein derivatives by P450 oxidation systems and a map of the constructed plasmid pGAP-CYP-CPR. |
| 關鍵字： | Daidzein, Pichia, Biotransformation |
| 著作名稱： | Knockdown of Adenosine Deaminase 1 Acting on RNA Inhibits Expression of Proopiomelanocortin in HaCaT Cells |
| 年度： | 2012 |
| 類別： |
會議論文
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| 摘要： | In 2003, ADAR1 (Adenosine Deaminase 1 Acting on RNA) was identified as the disease gene for an autosomal dominant disease — dyschromatosis symmetrica hereditaria (DSH) — which is characterized by a mixture of hyperpigmented and hypopigmented macules that are distributed on the dorsal aspects of the extremities and freckle-like macules on the face. To date, more than 100 ADAR1 mutations have been discovered in DSH patients. However, the exact molecular pathogenesis of DSH has not been clearly identified. To investigate the effects of ADAR1 mutations on the signal transduction pathway of melanogenesis, we used the RNA interference (RNAi) method to knockdown ADAR1 in human keratinocyte HaCaT cells, and determined a hormone precursor peptide proopiomelanocortin (POMC) expression in the ADAR1-knockdown HaCaT cells. Based on the results, we found that ADAR1 knockdown inhibits POMC expression in HaCaT cells. The findings of the present study imply that ADAR1 plays an important role in regulating POMC expression in keratinocytes, and possibly, the UV-induced pigmentation of skin. |
| 關鍵字： | ADAR1, HaCaT, RNA editing |
| 著作名稱： | Anti-melanoma activity of corylin from Psoralea corylifolia |
| 年度： | 2023 |
| 類別： |
會議論文
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| 摘要： | |
| 關鍵字： | |
| 著作名稱： | Using Biotransformation-guided Purification to Produce New Ganoderma Saponins |
| 年度： | 2023 |
| 類別： |
會議論文
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| 摘要： | |
| 關鍵字： | |
| 著作名稱： | Production of soy isoflavone glycosides 3-hydroxydaidzin and 3-hydroxygenistin by using recombinant Escherichia coli |
| 年度： | 2016 |
| 類別： |
會議論文
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| 摘要： | The present study describes the biotransformation of a commercially available crude extract of soy isoflavones, which contained significant amounts of the soy isoflavone glycosides daidzin and genistin, by recombinant Escherichia coli expressing tyrosinase from Bacillus megaterium. Two major products were isolated from the biotransformation and identified as 3’-hydroxydaidzin and 3’-hydroxygenistin, respectively, based on their mass and nuclear magnetic resonance spectral data. |
| 關鍵字： | 3-hydroxydaidzin, 3-hydoxygenistin |
| 著作名稱： | PRODUCTION AND ANTI-TYROSINASE ACTIVITY OF METHOXY-QUERCETIN |
| 年度： | 2016 |
| 類別： |
會議論文
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| 摘要： | Abstract –The gene encoding O-methyltransferase from Streptomyces peucetius was subcloned into pETDuet-1TM to form the expression vector pETDuet-SpOMT2884. Biotransformation of quercetin by the recombinant Escherichia coli harboring the expression vector was conducted. The results showed that the recombinant strain catalyzed biotransformation of quercetin. The produced methoxy-quercetin was then isolated by preparative high performance liquid chromatography method. The inhibitory activity of the purified methoxy-quercetin on tyrosinase activity was determined and the results showed that purified methoxy-quercetin potently inhibited tyrosinase activity above 25 M. The present study highlights the applications of the production and anti-tyrosinase activity of the methoxy-quercetin in the cosmetics industry.
Keywords: Methoxy-quercetin, tyrosinase, inhibition
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| 關鍵字： | Methoxy-quercetin, tyrosinase, inhibition |
| 著作名稱： | BIOTRANSFORMATION OF 3’-HYDROXYGENISTEIN BY RECOMBINANT ESCHERICHIA COLI EXPRESSING O-METHYLTRANSFERASE |
| 年度： | 2016 |
| 類別： |
會議論文
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| 摘要： | Recombinant Escherichia coli was constructed to express O-methyltransferase (OMT) SpOMT2884 from Streptomyces peucetius. Biotransformation of 3’-hydroxygenistein by the recombinant E. coli was investigated. The results showed that two metabolites were produced using ultra-pressure liquid chromatography (UPLC) analysis. The two methoxy-isoflavones metabolites were isolated by using preparative high-pressure liquid chromatography (HPLC). Anti-melanogenesis activity of both methoxy-isoflavones metabolites were determined using cultured B16 melanoma cells. The results showed that unlike the precursor 3’-hydroxygenistein, which showed dose-dependent inhibitory effects on melanogenesis, the two metabolites showed no significantly inhibitory effects on melanogenesis. |
| 關鍵字： | Genistien, Daidzein, Hydoxylation |
| 著作名稱： | Identification of 3-hydroxygenistein as a potent melanogenesis inhibitor from biotransformation of genistein by recombinant Pichia pastoris |
| 年度： | 2015 |
| 類別： |
會議論文
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| 摘要： | A product resulting from the biotransformation of genistein by a recombinant Pichia pastoris was isolated and identified as 3-hydroxygenistein, on the basis of mass, 1H-NMR, and 13C-NMR spectrophotometric analysis. The maximal product concentration and the conversion yield of the biotransformation in a 5 l fermenter were 3.5 mg/l and 14%, respectively. The inhibitory effects of 3-hydroxygenistein on tyrosinase activity were investigated in vitro using mushroom tyrosinase. The results showed that 3-hydroxygenistein potently inhibited tyrosinase activity with an IC50 value of 15.9 M. Furthermore, the inhibitory effects of 3-hydroxygenistein on melanogenesis were also investigated in vitro in cultured B16 melanoma cells, and it was shown that 3-hydroxygenistein dose-dependently inhibited melanogenesis in non-toxic concentrations. In summary, the 3-hydroxygenistein that was produced from genistein by the recombinant yeast was confirmed as a potent tyrosinase inhibitor and inhibited melanogenesis in B16 cells. |
| 關鍵字： | Genistein, Melanogenesis, Inhibition |
| 著作名稱： | Improving 3-hydroxygenistein production in recombinant Pichia pastoris via adaptive laboratory evolution |
| 年度： | 2015 |
| 類別： |
會議論文
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| 摘要： |
3-hydroxygenistein can be from biotransformation of genistein by engineered Pichia pastoris X-33 harboring a fusion gene composed of CYP57B3 from Aspergillus oryzae, and a cytochrome reductase gene from Saccharomyces cerevisiae. Herein, adaptive laboratory evolution was further applied to select higher 3-hydroxygenistein production strains from previous P. pastoris X-33 using periodic hydrogen peroxide-shocking strategy. One hyper-producing strain, P2-D14-5, evolved to produce 23.0 mg/l of 3-hydroxygenistein, which is 1.87-fold higher than that produced by the original strain, and is the highest reported to date. Results from cell growth and real time qRT-PCR analysis showed that the evolved mutations that led to the improvement in production were not directly related to the cell growth and expression of the heterogeneous recombinant genes. In a scale-up experiment, P2-D14-5 produced 20.3 mg/l of 3-hydroxygenistein in a 5-L fermenter. The evolved P2-D14-5 strain has the potential for 3-hydroxygenistein production in industry.
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| 關鍵字： | Genistein, improvement, laboratory evolution |
| 著作名稱： | Inhibition of Melanogenesis by Yeast Extracts from Cultivations of Recombinant Pichia pastoris Catalyzing ortho-Hydroxylation of Flavonoids |
| 年度： | 2014 |
| 類別： |
會議論文
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| 摘要： | 摘要(Abstract)
The inhibition of melanogenesis by yeast extracts from cultivations of recombinant Pichia pastoris catalyzing ortho-hydroxylation of flavonoids was investigated. The recombinant yeast harbored a fusion gene composed of the CYP57B3 gene from Aspergillus oryzae and a cytochrome reductase gene from Saccharomyces cerevisiae. The results showed that five flavonoids, including the flavone apigenin, the flavanones naringenin and liquiritigenin, and the isoflavones daidzein and genistein, could be biotransformed. The yeast extracts from the five biotransformation fermentations were then evaluated for inhibitory activity on melanogenesis in cultured mouse B16 melanoma cells. The results demonstrate that the yeast extract from genistein biotransformation possessed the highest inhibitory activity.
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| 關鍵字： | Melanogenesis, Inhibition, Flavonoids, Hydroxylation, Pichia pastoris |