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Enhanced production of glycyrrhetic acid 3-O-mono-β-D-glucuronide by fed-batch fermentation using p H and dissolved oxygen as feedback parameters 被引量:2
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作者 Bo Lu Xiaogang Yang +1 位作者 Xudong Feng Chun Li 《Chinese Journal of Chemical Engineering》 SCIE EI CAS CSCD 2016年第4期506-512,共7页
Glycyrrhetic acid 3-O-mono-β-D-glucuronide (GAMG), the major functional ingredient in licorice, has widespread applications in food, pharmacy and cosmetics industry. The production of GAMG through Penicillium purpu... Glycyrrhetic acid 3-O-mono-β-D-glucuronide (GAMG), the major functional ingredient in licorice, has widespread applications in food, pharmacy and cosmetics industry. The production of GAMG through Penicillium purpurogenum Li-3 cultivation was for the first time performed through both batch and fed-batch processes in bioreactors. In batch process, under optimal conditions (pH 5.0, temperature 32℃, agitation speed 100 r. rain 1), 3.55 g. L^-1 GAMG was obtained in a 2.5 L fermentor. To further enhance GAMG production, a fine fed-batch process was developed by using pH and DO as feedback parameters. Starting from 48 h, 100 m190 g-L 1 substrate Glycyrrhizin (GL) was fed each time when pH increased to above 5.0 and DO was increased to above 80%. This strategy can significantly enhance GAMG production: the achieved GL conversion was 95.34% with GAMG yield of 95.15%, and GAMG concentration was 16.62 g. L^-1 which was 5 times higher than that of batch. Then, a two-step separation strat- egy was established to separate GAMG from fermentation broth by crude extraction of 15 ml column packed with D10I resin followed by fine purification with preparative C18 chromatography. The obtained GAMG purity was 95.79%. This study provides a new insight into the industrial bioprocess of high-level GAMG production. 展开更多
关键词 glycyrrhetic acid 3-o-mono-β -d-glucuronideFed-batch fermentationPenicillium purpurogenum Li-3D1 O1 resin separation
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O-glycosyltransferases from Homo sapiens contributes to the biosynthesis of Glycyrrhetic Acid 3-O-mono-β-D-glucuronide and Glycyrrhizin in Saccharomyces cerevisiae 被引量:6
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作者 Ke Xu Yu-jia Zhao +5 位作者 Nadeem Ahmad Jing-nan Wang Bo Lv Ying Wang Jun Ge Chun Li 《Synthetic and Systems Biotechnology》 SCIE 2021年第3期173-179,共7页
Glycyrrhizin(GL)and Glycyrrhetic Acid 3-O-mono-β-D-glucuronide(GAMG)are the typical triterpenoid glycosides found in the root of licorice,a popular medicinal plant that exhibits diverse physiological effects and phar... Glycyrrhizin(GL)and Glycyrrhetic Acid 3-O-mono-β-D-glucuronide(GAMG)are the typical triterpenoid glycosides found in the root of licorice,a popular medicinal plant that exhibits diverse physiological effects and pharmacological manifestations.However,only few reports are available on the glycosylation enzymes involved in the biosynthesis of these valuable compounds with low conversion yield so far.In mammals,glycosyltransferases are involved in the phase II metabolism and may provide new solutions for us to engineer microbial strains to produce high valued compounds due to the substrate promiscuity of these glycosyltransferases.In this study,we mined the genomic databases of mammals and evaluated 22 candidate genes of O-glycosyltransferases by analyzing their catalytic potential for O-glycosylation of the native substrate,glycyrrhetinic acid(GA)for its glycodiversification.Out of 22 selected glycosyltransferases,only UGT1A1 exhibited high catalytic performance for biosynthesis of the key licorice compounds GL and GAMG.Molecular docking results proposed that the enzymatic activity of UGT1A1 was likely owing to the stable hydrogen bonding interactions and favorite conformations between the amino acid residues around substrate channels(P82~R85)and substrates.Furthermore,the complete biosynthesis pathway of GL was reconstructed in Saccharomyces cerevisiae for the first time,resulting in the production of 5.98±0.47 mg/L and 2.31±0.21 mg/L of GL and GAMG,respectively. 展开更多
关键词 O-glycosyltransferase Homo sapiens Glycyrrhizin(GL) Saccharomyces cerevisiae glycyrrhetic acid 3-o-mono-β-d-glucuronide(GAMG)
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Construction of a CaHP04-PGUSl hybrid nanoflower through protein-inorganic self-assembly, and its application in glycyrrhetinic acid 3-0-mono-β-D-glucuronide preparation 被引量:1
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作者 Tian Jiang Yuhui Hou +2 位作者 Tengjiang Zhang Xudong Feng Chun Li 《Frontiers of Chemical Science and Engineering》 SCIE EI CAS CSCD 2019年第3期554-562,共9页
Glycyrrhetinic acid 3-0-mono-β-D-glucuronide (GAMG), an important pharmaceutical intermediate and functional sweetener, has broad applications in the food and medical industries. A green and cost-effective method for... Glycyrrhetinic acid 3-0-mono-β-D-glucuronide (GAMG), an important pharmaceutical intermediate and functional sweetener, has broad applications in the food and medical industries. A green and cost-effective method for its preparation is highly desired. Using sitedirected mutagenesis, we previously obtained a variant of β-glucuronidase from Aspergillus oryzae Li-3 (PGUS1), which can specifically transform glycyrrhizin (GL) into GAMG. In this study, a facile method was established to prepare a CaHP04-PGUSl hybrid nanoflower for enzyme immobilization, based on protein-inorganic hybrid selfassembly. Under optimal conditions, 1.2 mg of a CaHP04- PGUS1 hybrid nanoflower precipitate with 71.2% immobilization efficiency, 35.60 mg·g^-1 loading capacity, and 118% relative activity was obtained. Confocal laser scanning microscope and scanning electron microscope results showed that the enzyme was encapsulated in the CaHP04-PGUSl hybrid nanoflower. Moreover, the thermostability of the CaHP04-PGUSl hybrid nanoflower at 55°C was improved, and its half-life increased by 1.3 folds. Additionally, the CaHP04-PGUSl hybrid nanoflower was used for the preparation of GAMG through GL hydrolysis, with the conversion rate of 92% in 8 h, and after eight consecutive runs, it had 60% of its original activity. 展开更多
关键词 β-glucuronidase enzyme-inorganic hybrid nanoflower biotransformation glycyrrhizin glycyrrtinic acid 3-o-mono-β-d-glucuronide
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