國立臺南大學教師基本資料

基本資料
姓名 張德生
系所 生物科技學系
職稱 教授
校內分機 7310、7723
傳真 (06)2602137
辦公室/研究室 ZA306-1、ZE301-2
E-mail cts@mail.nutn.edu.tw
網址
專長/研究領域 微生物生技產品開發、異黃酮多酚衍生物量產與美白化妝品開發
 

畢業學校國別主修學門學位修業期間
清華大學中華民國生科系學士1991-1995
清華大學中華民國生科系碩士1995-1997
清華大學中華民國化工系博士1999-2003

服務機關部門 / 系所職稱服務期間
嘉南科技大學化妝品系助理教授2003-2004
臺南大學生科系助理教授2004-2007
臺南大學生科系副教授2007-2010
臺南大學生科系教授2010~
臺南大學生科系系主任2011-2017

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
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
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
摘要
關鍵字
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
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
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
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
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].
年度2007
類別期刊論文
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.
關鍵字Keywords: Inhibitor; Isoflavone; Irreversible; Soygerm; Tyrosinase
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
摘要
關鍵字
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
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
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
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:
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
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.
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
名稱Melanogenesis inhibition by homoisoflavanone sappanone A from Caesalpinia sappan.
年度2012
類別期刊論文
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
名稱Natural Melanogenesis Inhibitors Acting Through the Down-Regulation of Tyrosinase Activity.
年度2012
類別期刊論文
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
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
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
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
名稱Isolation, bioactivity, and production of ortho-hydroxydaidzein and ortho-hydroxygenistein.
年度2014
類別期刊論文
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
名稱Methanol Partition Fraction of Ethanol of Discorea nipponica Makino Inhibits Melanogenesis
年度2014
類別期刊論文
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
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
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
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
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
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
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
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
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
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
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?
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
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
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
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
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
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
名稱Knockdown of Adenosine Deaminase 1 Acting on RNA Inhibits Expression of Proopiomelanocortin in HaCaT Cells
年度2012
類別研討會
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
名稱TRANSFORMATION OF DAIDZEIN BY A RECOMBINANT STRAIN OF PICHIA PASTORIS
年度2013
類別研討會
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
名稱Production of 6-Hydroxyapigenin by Recombinant Pichia pastoris Harboring Fusion P450 Gene
年度2014
類別研討會
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.
關鍵字6-Hydroxyapigenin, Pichia, P450
名稱Inhibition of Melanogenesis by Yeast Extracts from Cultivations of Recombinant Pichia pastoris Catalyzing ortho-Hydroxylation of Flavonoids
年度2014
類別研討會
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.
關鍵字Melanogenesis, Inhibition, Flavonoids, Hydroxylation, Pichia pastoris
名稱Identification of 3-hydroxygenistein as a potent melanogenesis inhibitor from biotransformation of genistein by recombinant Pichia pastoris
年度2015
類別研討會
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
名稱Improving 3-hydroxygenistein production in recombinant Pichia pastoris via adaptive laboratory evolution
年度2015
類別研討會
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.
關鍵字Genistein, improvement, laboratory evolution
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
類別研討會
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.
關鍵字CYP105D7, isoflavones, daidzein, genistein, Streptomyces
名稱Biotransformation of Quercetin by Recombinant Escherichia coli Expressing O-Methyltransferase from Streptomyces peucetius
年度2016
類別研討會
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
名稱Biotransformation of 3’-Hydroxydaidzein by Escherichia coli Expressing O-Methyltransferase
年度2016
類別研討會
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
名稱PRODUCTION AND ANTI-TYROSINASE ACTIVITY OF METHOXY-QUERCETIN
年度2016
類別研討會
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
關鍵字 Methoxy-quercetin, tyrosinase, inhibition
名稱BIOTRANSFORMATION OF 3’-HYDROXYGENISTEIN BY RECOMBINANT ESCHERICHIA COLI EXPRESSING O-METHYLTRANSFERASE
年度2016
類別研討會
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
名稱Production of soy isoflavone glycosides 3-hydroxydaidzin and 3-hydroxygenistin by using recombinant Escherichia coli
年度2016
類別研討會
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
名稱Inhibitory Effects of 8-Methoxydaidzein on Tyrosinase Activity in Mouse B16 Melanoma Cells
年度2017
類別研討會
摘要Our previous study has
關鍵字Daidzein, 8-hydroxydaidzein, methylation, methyltransferase, melanogenesis, inhibition, tyrosinase