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一种利用甘氨酸氧化酶生产乙醛酸的方法
引用本文:王玉成,宋伟,艾连中,吴静. 一种利用甘氨酸氧化酶生产乙醛酸的方法[J]. 上海理工大学学报, 2021, 43(1): 42-48. DOI: 10.13255/j.cnki.jusst.20200221002
作者姓名:王玉成  宋伟  艾连中  吴静
作者单位:江南大学药学院,江苏无锡214122;上海理工大学医疗器械与食品学院,上海200093
基金项目:国家轻工技术与工程一流学科自主课题(LITE2018-20)
摘    要:为实现生物法工业化生产乙醛酸,采用基因工程技术对甘氨酸氧化酶(ThiO)进行表达优化,使用SDS-PAGE电泳手段验证ThiO的可溶性表达,通过全细胞转化法检测重组菌体生产乙醛酸的能力,并优化了反应条件,在放大体系中考察其应用性。结果表明:构建的菌株Bacillus subtilis 168(pP43NMK-ThiO)...

关 键 词:甘氨酸  甘氨酸氧化酶  可溶性表达  全细胞催化  乙醛酸
收稿时间:2020-02-21

Production of glyoxylic acid by glycine oxidase
WANG Yucheng,SONG Wei,AI Lianzhong,WU Jing. Production of glyoxylic acid by glycine oxidase[J]. Journal of University of Shanghai For Science and Technology, 2021, 43(1): 42-48. DOI: 10.13255/j.cnki.jusst.20200221002
Authors:WANG Yucheng  SONG Wei  AI Lianzhong  WU Jing
Affiliation:School of Pharmaceutical Sciences, Jiangnan University, Wuxi 214122, China;School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China
Abstract:In order to realize the industrial production of glyoxylic acid, the expression of glycine oxidase (ThiO) was optimized using genetic engineering technology. The soluble expression of recombinant strains was verified by SDS-PAGE electrophoresis. The ability of recombinant bacteria to produce glyoxylic acid was tested by whole cell transformation. With the optimization of reaction conditions, its applicability in the enlarged transformation system was investigated. The strain Bacillus subtilis 168 (pP43NMK-ThiO) achieved intracellular soluble expression and the yield of glyoxylic acid from glycine by the whole cell transformation surpassed the highest yield of glyoxylic acid produced by current biological methods. The best reaction conditions were 30 ℃ and pH value 8.3. The whole cell catalyst maintained high thermal stability at 35 ℃, and more than still maintains more than reaction activity after 8 h incubation. Under the optimal reaction conditions, the yield was further increased to 30% by adding catalase at a final concentration of 1%. The results from the 3 L fermentation tank are consistent with those obtained from the shake flask, showing stable conversion capacity. The results lays the foundation for the industrialization of the preparation of glyoxylic acid by the biological method.
Keywords:glycine  glycine oxidase  soluble expression  whole cell catalysis  glyoxylic acid
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