Recombinant Expression and Characterization of Ornithine Transcarbamoylase from Lactobacillus brevis

被引：0

作者：

Fang R. ^{[1
,2
]}

Zhou C. ^{[1
,2
]}

Jin C. ^{[1
,2
]}

Zhang Y. ^{[1
,2
]}

Huang W. ^{[1
,2
]}

Huang J. ^{[1
,2
]}

Lü C. ^{[1
,2
]}

Xiao G. ^{[1
,2
]}

Gong J. ^{[1
,2
]}

Chen Q. ^{[3
]}

机构：

[1] Zhejiang University of Science and Technology, Hangzhou

[2] Zhejiang Provincial Key Laboratory of Agricultural Products Chemistry and Bioprocessing Technology, Hangzhou

[3] Zhejiang University, Hangzhou

来源：

Journal of Chinese Institute of Food Science and Technology | 2020年 / 20卷 / 11期

关键词：

Cloning; Enzyme properties; Expression; Ornithine transcarbamoylase; Purification;

D O I：

10.16429/j.1009-7848.2020.11.010

中图分类号：

学科分类号：

摘要：

Objective: To clone and recombinantly express ornithine transcarbamoylase from Lactobacillus brevis and study its enzymatic properties. Methods: The gene cloning process was using Lactobacillus brevis as template which based on the gene sequence analysis. The recombinant protein was expressed in E. coli, and purified by Ni-NTA affinity chromatography column. By using arginine as substrate, the enzymatic properties were studied. Results: The optimal temperature of recombinant ornithine transcarbamoylase is 55℃. It has excellent thermal stability at 4℃ and still retains more than 50% biological activity at 37℃ for 3 hours. However, after 40 minutes at 65℃ it will totally lose activity; the optimal pH value of the enzyme is 6.5, when the pH is alkaline, the enzyme activity decreases significantly; the enzyme possess highest activity under 5% mass fraction of NaCl and 5% volume fraction of ethanol, but the difference is not obvious, indicating recombinant ornithine transcarbamoylase has good salt tolerance and alcohol tolerance; the kinetic parameters Km is 1.58 mmol and Vmax is 5.88 mmol/min. Conclusion: The ornithine transcarbamoylase derived from Lactobacillus brevis has good temperature stability, salt tolerance and alcohol tolerance, and has certain application prospects in alcohol fermentation. © 2020, Editorial Office of Journal of CIFST. All right reserved.

引用

页码：79 / 84

页数：5

共 17 条

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