甘草根提取物中甘草酸的筛选及不同微生物菌株对18β-甘草次酸的生物转化

Makhmur Ahmad, Jalaluddin, M. Ali, B. Panda
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摘要

目的:筛选不同微生物(7种细菌和14种真菌)将甘草酸(GL)转化为18β-甘草酸(GA)。青霉菌产β-葡萄糖醛酸酶最高(61 U/mL), GA浓度为52 μg/mL;大肠杆菌产β-葡萄糖醛酸酶最高(376 U/mL), GA浓度为2.1 μg/mL。材料与方法:对GL进行了浸没和固态生物转化。取细菌上清9.0 mL,加入0.2 w/v甘草根提取物1.0 mL, 37℃孵育24 h,测定β-葡萄糖醛酸酶活性并进行高效液相色谱分析。结果与讨论:诱导大肠杆菌产生2.1 μg/mL的GA,酶活性为376 U/mL,表明该酶具有潜在的生物转化能力。米根霉(Rhizopus oryzae)和黄根霉(P. chrysogenum)分别以2.6 μg/mL和61 μg/mL的GA酶活性将GL转化为GA,酶活性分别为569 U/mL和61 U/mL。结论:微生物β-葡萄糖醛酸酶可在水下发酵(SmF)条件下水解含GL的光草根。结果表明,在SmF条件下,大肠杆菌β-葡萄糖醛酸酶是比真菌菌株更好的生物催化酶源。
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Screening and bioconversion of glycyrrhizin of Glycyrrhiza glabra root extract to 18β-glycyrrhetinic acid by different microbial strains
Objective: The objective of the present study is to perform screening of different microorganisms (7 bacteria and 14 fungi) for conversion of glycyrrhizin (GL) to 18β-glycyrrhetinic acid (GA). Penicillium chrysogenum produced the highest concentration of β-glucuronidase enzyme (61 U/mL) and produced GA of 52 μg/mL while E. coli produced the highest β-glucuronidase of 376 U/mL with GA concentration of 2.1 μg/mL. Materials and Methods: Submerged and solid state biotransformation of GL was carried out. To 9.0 mL of bacterial supernatant, 1.0 mL 0.2% w/v of aqueous Glycyrrhiza glabra root extract was added and incubated at 37°C for 24 h. β-glucuronidase activity was measured and high-performance liquid chromatography analysis was carried out. Results and Discussion: Induced-Escherichia coli produces 2.1 μg/mL of GA with an enzyme activity of 376 U/mL which shows that the enzyme has a potential biotransformation capability. Rhizopus oryzae and P. chrysogenum have the potential ability to biotransform GL to GA with 2.6 μg/mL and 61 μg/mL of GA with enzyme activity of 569 U/mL and 61 U/mL, respectively. Conclusions: G. glabra roots containing GL can be hydrolyzed by microbial β-glucuronidase enzyme under sub-merged fermentation (SmF). β-glucuronidase, an enzyme of E. coli, was found to be the best microbial source of enzyme which biocatalyzed the reaction than fungal strain under SmF.
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