Heterologous expression and enzymatic characterization of glucosamine-6-phosphate synthase

被引：0
|
作者：
Wang T. [1 ]
Feng J.-R. [1 ]
Li S.-X. [1 ]
Du X. [1 ]
Liu X.-H. [1 ]
Wu M.-B. [2 ]
机构：
[1] School of Biological Science, Jining Medical University, Rizhao
[2] Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou
来源：
Gao Xiao Hua Xue Gong Cheng Xue Bao/Journal of Chemical Engineering of Chinese Universities | 2021年 / 35卷 / 03期
关键词：
Antifungal target; Enzymatic characterization; Glucosamine-6-phosphate synthase; Heterologous expression;
D O I：
10.3969/j.issn.1003-9015.2021.03.015
中图分类号：
学科分类号：
摘要：
Glucosamine-6-phosphate synthase (GFA) is a key enzyme in the biosynthesis of hexosamine. In order to establish an efficient GFA production process, the heterologous expression of GFA from pathogenic bacterium Candida albicans ATCC 64548 was expressed in E. coli BL21(DE3) Codon plus. The results show that the optimal expression conditions of GFA were: the concentration of IPTG 0.1 mmol∙L-1; induction temperature 28 ℃; induction time 15 h; induction timing (OD600) reaching 1.0. The optimum reaction conditions were determined as: reaction temperature 37 ℃; reaction pH 7.2, and reaction time 20 min. The KM value of GFA was determined as 0.47 mg∙mL-1 by Lineweaver-Burk. When the inhibitor (bacilysin) was added, the transaminase activity of GFA was significantly inhibited and reduced by 54.8 %. With the method developed in this study, glucosamine-6-phosphate synthase could be expressed heterologously, which would greatly increase its application and help accelerate the development of new antifungal drugs. © 2021, Editorial Board of "Journal of Chemical Engineering of Chinese Universities". All right reserved.
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页码：500 / 506
页数：6
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