CN110387360A - 羟基类固醇脱氢酶及其在合成熊去氧胆酸前体中的应用 - Google Patents
羟基类固醇脱氢酶及其在合成熊去氧胆酸前体中的应用 Download PDFInfo
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- CN110387360A CN110387360A CN201910527373.7A CN201910527373A CN110387360A CN 110387360 A CN110387360 A CN 110387360A CN 201910527373 A CN201910527373 A CN 201910527373A CN 110387360 A CN110387360 A CN 110387360A
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- hydroxysteroid dehydrogenase
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Cited By (8)
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CN110760488A (zh) * | 2019-12-12 | 2020-02-07 | 四川澄华生物科技有限公司 | 一种高含量12α-羟基类固醇脱氢酶发酵液的发酵方法 |
CN111593085A (zh) * | 2020-05-26 | 2020-08-28 | 四川澄华生物科技有限公司 | 一种12-酮基胆酸的制备方法 |
CN113151207A (zh) * | 2021-04-21 | 2021-07-23 | 重庆第二师范学院 | NAD(H)依赖型3α-羟基类固醇脱氢酶及其编码基因 |
CN113604446A (zh) * | 2021-08-16 | 2021-11-05 | 重庆大学 | 7α-羟基类固醇脱氢酶St-2-2的突变体R16Q |
CN114107237A (zh) * | 2021-09-07 | 2022-03-01 | 伊犁川宁生物技术股份有限公司 | 一种发酵生产鹅去氧胆酸氧化酶的方法 |
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CN114317663A (zh) * | 2022-01-19 | 2022-04-12 | 常德云港生物科技有限公司 | 利用猪胆提取胆红素后的下料合成熊去氧胆酸的方法 |
CN114854707A (zh) * | 2022-06-14 | 2022-08-05 | 苏州百福安酶技术有限公司 | 一种7β-羟基甾体脱氢酶突变体 |
Citations (21)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101421413A (zh) * | 2006-04-11 | 2009-04-29 | Iep有限责任公司 | 用羟基类固醇脱氢酶通过还原氧代类固醇化合物或者氧化羟基类固醇化合物制备类固醇衍生物的方法 |
CN102007210A (zh) * | 2008-03-26 | 2011-04-06 | 细胞制药有限公司 | 新的12α-羟类固醇脱氢酶,产生及其用途 |
CN102245779A (zh) * | 2008-12-11 | 2011-11-16 | 生物结构实验室公司 | 大宗化学品的生物合成 |
WO2012080504A1 (de) * | 2010-12-16 | 2012-06-21 | Pharmazell Gmbh | NEUARTIGE 7ß-HYDROXYSTEROID DEHYDROGENASE-MUTANTEN UND VERFAHREN ZUR HERSTELLUNG VON URSODESOXYCHOLSÄURE |
NZ603816A (en) * | 2010-05-27 | 2014-06-27 | Pharmazell Gmbh | Novel 7alpha-hydroxysteroid dehydrogenase knockout mutants and use thereof |
CN104136620A (zh) * | 2012-02-07 | 2014-11-05 | 安尼基有限责任公司 | 用于氧化还原辅因子的酶再生的方法 |
CN104673765A (zh) * | 2014-12-22 | 2015-06-03 | 武汉康复得生物科技股份有限公司 | 一组在生理pH条件下有活力的草酸氧化酶及其应用 |
WO2015197698A2 (de) * | 2014-06-24 | 2015-12-30 | Pharmazell Gmbh | NEUARTIGE VERFAHREN ZUR BIOKATALYTISCHEN GANZZELLREDUKTION VON DEHYDROCHOLSÄUREVERBINDUNGEN, NEUARTIGE 7ß-HYDROXYSTEROID DEHYDROGENASE-MUTANTEN UND VERBESSERTE BIOKATALYTISCHE VERFAHREN ZUR HERSTELLUNG VON URSODESOXYCHOLSÄURE |
WO2016023933A1 (de) * | 2014-08-12 | 2016-02-18 | Pharmazell Gmbh | 3alpha-hydroxysteroid dehydrogenase-mutanten und verfahren zur herstellung von ursodesoxycholsäure |
CN106282138A (zh) * | 2016-09-22 | 2017-01-04 | 重庆大学 | 撒丁岛梭菌7α‑羟基类固醇脱氢酶突变体T145S |
CN106434582A (zh) * | 2016-09-22 | 2017-02-22 | 重庆大学 | 撒丁岛梭菌7α‑羟基类固醇脱氢酶突变体R194A |
CN106701882A (zh) * | 2017-01-24 | 2017-05-24 | 尚科生物医药(上海)有限公司 | 化学‑酶法制备熊去氧胆酸 |
WO2017123592A8 (en) * | 2016-01-11 | 2017-08-31 | Synlogic, Inc. | Bacteria engineered to treat disorders associated with bile salts |
CN107384820A (zh) * | 2017-07-25 | 2017-11-24 | 华东理工大学 | 一株谷氨酰胺转氨酶高产诱变菌株及其应用 |
CN107418923A (zh) * | 2017-09-20 | 2017-12-01 | 华东理工大学 | 伯克霍尔德菌及其应用 |
WO2017220486A2 (en) * | 2016-06-20 | 2017-12-28 | Pharmazell Gmbh | Coupled, self-sufficient biotransformation of chenodeoxycholic acid to ursodeoxycholic acid and novel enzyme mutants applicable in said process |
CN108546691A (zh) * | 2018-05-09 | 2018-09-18 | 华东理工大学 | 7β-羟基甾醇脱氢酶突变体及其在制备熊脱氧胆酸中的应用 |
CN109055473A (zh) * | 2018-08-21 | 2018-12-21 | 湖南宝利士生物技术有限公司 | 一种基于酶法偶联技术合成熊去氧胆酸和高手性纯度d-氨基酸的方法 |
CN109182284A (zh) * | 2018-09-28 | 2019-01-11 | 湖南福来格生物技术有限公司 | 一种7β-羟基类固醇脱氢酶突变体、编码序列、重组表达载体、基因工程菌及应用 |
CN109355266A (zh) * | 2018-11-06 | 2019-02-19 | 华东理工大学 | 亚胺还原酶突变体及其在光学活性1-取代-四氢异喹啉衍生物合成中的应用 |
CN109722463A (zh) * | 2019-03-18 | 2019-05-07 | 常德云港生物科技有限公司 | 一种通过半酶法将猪胆酸合成熊去氧胆酸的方法 |
-
2019
- 2019-06-18 CN CN201910527373.7A patent/CN110387360B/zh active Active
Patent Citations (23)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101421413A (zh) * | 2006-04-11 | 2009-04-29 | Iep有限责任公司 | 用羟基类固醇脱氢酶通过还原氧代类固醇化合物或者氧化羟基类固醇化合物制备类固醇衍生物的方法 |
CN102007210A (zh) * | 2008-03-26 | 2011-04-06 | 细胞制药有限公司 | 新的12α-羟类固醇脱氢酶,产生及其用途 |
CN102245779A (zh) * | 2008-12-11 | 2011-11-16 | 生物结构实验室公司 | 大宗化学品的生物合成 |
NZ603816A (en) * | 2010-05-27 | 2014-06-27 | Pharmazell Gmbh | Novel 7alpha-hydroxysteroid dehydrogenase knockout mutants and use thereof |
CN105441399A (zh) * | 2010-12-16 | 2016-03-30 | 细胞制药有限公司 | 新的7β-羟基类固醇脱氢酶突变体和制备熊去氧胆酸的方法 |
WO2012080504A1 (de) * | 2010-12-16 | 2012-06-21 | Pharmazell Gmbh | NEUARTIGE 7ß-HYDROXYSTEROID DEHYDROGENASE-MUTANTEN UND VERFAHREN ZUR HERSTELLUNG VON URSODESOXYCHOLSÄURE |
JP2017079788A (ja) * | 2010-12-16 | 2017-05-18 | ファルマツェル、ゲーエムベーハー | 新規7β−ヒドロキシステロイドデヒドロゲナーゼ変異体及びウルソデオキシコール酸の調製方法 |
CN104136620A (zh) * | 2012-02-07 | 2014-11-05 | 安尼基有限责任公司 | 用于氧化还原辅因子的酶再生的方法 |
WO2015197698A2 (de) * | 2014-06-24 | 2015-12-30 | Pharmazell Gmbh | NEUARTIGE VERFAHREN ZUR BIOKATALYTISCHEN GANZZELLREDUKTION VON DEHYDROCHOLSÄUREVERBINDUNGEN, NEUARTIGE 7ß-HYDROXYSTEROID DEHYDROGENASE-MUTANTEN UND VERBESSERTE BIOKATALYTISCHE VERFAHREN ZUR HERSTELLUNG VON URSODESOXYCHOLSÄURE |
WO2016023933A1 (de) * | 2014-08-12 | 2016-02-18 | Pharmazell Gmbh | 3alpha-hydroxysteroid dehydrogenase-mutanten und verfahren zur herstellung von ursodesoxycholsäure |
CN104673765A (zh) * | 2014-12-22 | 2015-06-03 | 武汉康复得生物科技股份有限公司 | 一组在生理pH条件下有活力的草酸氧化酶及其应用 |
WO2017123592A8 (en) * | 2016-01-11 | 2017-08-31 | Synlogic, Inc. | Bacteria engineered to treat disorders associated with bile salts |
WO2017220486A2 (en) * | 2016-06-20 | 2017-12-28 | Pharmazell Gmbh | Coupled, self-sufficient biotransformation of chenodeoxycholic acid to ursodeoxycholic acid and novel enzyme mutants applicable in said process |
CN106282138A (zh) * | 2016-09-22 | 2017-01-04 | 重庆大学 | 撒丁岛梭菌7α‑羟基类固醇脱氢酶突变体T145S |
CN106434582A (zh) * | 2016-09-22 | 2017-02-22 | 重庆大学 | 撒丁岛梭菌7α‑羟基类固醇脱氢酶突变体R194A |
CN106701882A (zh) * | 2017-01-24 | 2017-05-24 | 尚科生物医药(上海)有限公司 | 化学‑酶法制备熊去氧胆酸 |
CN107384820A (zh) * | 2017-07-25 | 2017-11-24 | 华东理工大学 | 一株谷氨酰胺转氨酶高产诱变菌株及其应用 |
CN107418923A (zh) * | 2017-09-20 | 2017-12-01 | 华东理工大学 | 伯克霍尔德菌及其应用 |
CN108546691A (zh) * | 2018-05-09 | 2018-09-18 | 华东理工大学 | 7β-羟基甾醇脱氢酶突变体及其在制备熊脱氧胆酸中的应用 |
CN109055473A (zh) * | 2018-08-21 | 2018-12-21 | 湖南宝利士生物技术有限公司 | 一种基于酶法偶联技术合成熊去氧胆酸和高手性纯度d-氨基酸的方法 |
CN109182284A (zh) * | 2018-09-28 | 2019-01-11 | 湖南福来格生物技术有限公司 | 一种7β-羟基类固醇脱氢酶突变体、编码序列、重组表达载体、基因工程菌及应用 |
CN109355266A (zh) * | 2018-11-06 | 2019-02-19 | 华东理工大学 | 亚胺还原酶突变体及其在光学活性1-取代-四氢异喹啉衍生物合成中的应用 |
CN109722463A (zh) * | 2019-03-18 | 2019-05-07 | 常德云港生物科技有限公司 | 一种通过半酶法将猪胆酸合成熊去氧胆酸的方法 |
Non-Patent Citations (7)
Title |
---|
MANFRED BRAUND等: ""12a-Hydroxysteroid dehydrogenase from Clostridium group P, strain C 48-50 Production, purification and characterization"", 《EUR. J. BIOCHEM.》 * |
NCBI: ""SDR family NAD(P)-dependent oxidoreductase [Rhodococcus ruber]"", 《GENBANK DATABASE》 * |
SEAN M. MYTHEN等: ""Targeted Synthesis and Characterization of a Gene Cluster Encoding NAD(P)H-Dependent 3α-, 3β-, and 12α- Hydroxysteroid Dehydrogenases from Eggerthella CAG:298, a Gut Metagenomic Sequence"", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 * |
SHOU-CHENG SHI等: ""Efficient Synthesis of 12-Oxochenodeoxycholic Acid Using a 12α-Hydroxysteroid Dehydrogenase from Rhodococcus ruber"", 《ADV. SYNTH. CATAL.》 * |
史杰等: ""牛磺熊去氧胆酸的获取方法及药理研究进展"", 《上海中医药大学学报》 * |
石守城: ""12α-羟基类固醇的基因挖掘及在合成12-oxo-鹅去氧胆酸中的研究"", 《中国生物工程学会第十三届学术年会暨2019年全国生物技术大会会议论文集》 * |
苏文等: ""17β-羟基类固醇脱氢酶的功能"", 《生理科学进展》 * |
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CN113151207A (zh) * | 2021-04-21 | 2021-07-23 | 重庆第二师范学院 | NAD(H)依赖型3α-羟基类固醇脱氢酶及其编码基因 |
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CN113604446A (zh) * | 2021-08-16 | 2021-11-05 | 重庆大学 | 7α-羟基类固醇脱氢酶St-2-2的突变体R16Q |
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CN114854707A (zh) * | 2022-06-14 | 2022-08-05 | 苏州百福安酶技术有限公司 | 一种7β-羟基甾体脱氢酶突变体 |
CN114854707B (zh) * | 2022-06-14 | 2023-09-12 | 苏州百福安酶技术有限公司 | 一种7β-羟基甾体脱氢酶突变体 |
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