克
化学
细菌
重组DNA
生物化学
微生物学
生物
遗传学
基因
作者
Dong-Shan An,Changhao Cui,Muḥammad Zubair Ṣiddiqi,Hong Shan Yu,Jin Feng-xie,Song-Gun Kim,Wan‐Taek Im
出处
期刊:Journal of Microbiology and Biotechnology
[Springer Science+Business Media]
日期:2017-09-28
卷期号:27 (9): 1559-1565
被引量:18
标识
DOI:10.4014/jmb.1703.03006
摘要
Naturally occurring ginsenoside F1 (20-O-β-D-glucopyranosyl-20(S)-protopanaxatriol) is rare. Here, we produced gram-scale quantities of ginsenoside F1 from a crude protopanaxatriol saponin mixture comprised mainly of Re and Rg1 through enzyme-mediated biotransformation using recombinant β-glucosidase (BgpA) cloned from a soil bacterium, Terrabacter ginsenosidimutans Gsoil 3082T. In a systematic step-by-step process, the concentrations of substrate, enzyme, and NaCl were determined for maximal production of F1. At an optimized NaCl concentration of 200 mM, the protopanaxatriol saponin mixture (25 mg/ml) was incubated with recombinant BgpA (20 mg/ml) for 3 days in a 2.4 L reaction. Following octadecylsilyl silica gel column chromatography, 9.6 g of F1 was obtained from 60 g of substrate mixture at 95% purity, as assessed by chromatography. These results represent the first report of gramscale F1 production via recombinant enzyme-mediated biotransformation.
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