Effects of chromosomal gene copy number and locations on polyhydroxyalkanoate synthesis by Escherichia coli and Halomonas sp

Applied Microbiology and Biotechnology
Jin YinGuo-Qiang Chen

Abstract

Chromosomal integration and expression of heterologous gene(s) are favored in industrial biotechnology due to the inheriting expression stability. Yet, chromosomal expression is commonly weaker than plasmid one. The effect on gene expression level at 13 chromosomal locations in Escherichia coli was investigated using the polyhydroxybutyrate (PHB) synthesis pathway encoded by a phaCAB operon as a reporter. When 11 copies of phaCAB were randomly integrated into 11 of the 13 chromosomal locations, respectively, 5.2 wt% of PHB was produced. PHB (34.1 wt%) was accumulated by a recombinant E. coli inserted chromosomally with 50 copies of phaCAB in the active asnB site using a Cre-loxP recombination method. This PHB accumulation level was equivalent to a medium-copy-number plasmid expression system, suggesting the importance of chromosomal gene copy number for PHB production by E. coli. This result was used to manipulate a Halomonas strain. One copy of genes scpAB encoding methylmalonyl-CoA mutase and methylmalonyl-CoA decarboxylase was inserted into the strongest expression site porin in the chromosome of the 2-methylcitrate synthase (prpC) deleted mutant Halomonas TD08, leading to the synthesis of poly(hydroxybutyrate-co-hydroxyvale...Continue Reading

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Citations

Nov 9, 2016·Journal of Industrial Microbiology & Biotechnology·Zheng-Jun LiGregory Stephanopoulos
Jun 18, 2017·Applied Microbiology and Biotechnology·Dan LiGuo-Qiang Chen
Dec 16, 2017·Applied Biochemistry and Biotechnology·Wei-Fan LiangXin-Hui Xing
Jul 8, 2018·Biotechnology and Bioengineering·Bingming OuGuoqiang Zhu
Jul 21, 2020·Biotechnology for Biofuels·Mohamed AmerNigel S Scrutton
Nov 6, 2020·Metabolic Engineering·Dongdong ZhaoXueli Zhang
Feb 14, 2021·Science Advances·Tatyana E SaleskiXiaoxia Nina Lin
Mar 18, 2021·Critical Reviews in Biotechnology·Kai-Hee HuongA A Amirul
Oct 15, 2021·Critical Reviews in Biotechnology·Pavni RekhiMousumi Debnath

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