Engineering of core promoter regions enables the construction of constitutive and inducible promoters in Halomonas sp

Biotechnology Journal
Tingting LiQi Ouyang

Abstract

Halomonas strain TD01, a newly identified halophilic bacterium, has proven to be a promising low-cost host for the production of chemicals. However, genetic manipulation in Halomonas sp. is still difficult due to the lack of well-characterized and tunable expression systems. In this study, a systematic, efficient method was exploited to construct both a constitutive promoter library and inducible promoters. Porin, a highly expressed protein in Halomonas TD01, was first identified from the Halomonas TD01 proteome. Subsequent study of the intergenic region upstream of porin led to the identification of a core promoter region, including -10 and -35 elements. By randomizing the sequence between the -35 and -10 elements, a constitutive promoter library was obtained with 310-fold variation in transcriptional activity; an inducible promoter with a >200-fold induction was built by integrating a lac operator into the core promoter region. As two complementary expression systems, the constitutive and inducible promoters were then employed to regulate the biosynthetic pathway of poly-3-hydroxybutyrate (PHB) in Halomonas TD01, demonstrating the usefulness of the expression systems, furthermore, they could be applied in future metabolic eng...Continue Reading

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Citations

Aug 21, 2016·Journal of Microbiological Methods·Joshua R HarrisChristopher T Nomura
Oct 21, 2019·Applied Microbiology and Biotechnology·Li-Qun JinYu-Guo Zheng
Jun 5, 2019·Genome Biology·Hengji ZhanYuchen Liu
Jun 28, 2018·Journal of Industrial Microbiology & Biotechnology·Xiangbin ChenGuo-Qiang Chen
Mar 7, 2020·Frontiers in Bioengineering and Biotechnology·Laichuang HanZhemin Zhou
Jul 21, 2020·Biotechnology for Biofuels·Mohamed AmerNigel S Scrutton
Jan 13, 2021·Trends in Biotechnology·Dan TanGuo-Qiang Chen
Dec 3, 2020·Synthetic Biology·Duangthip TrisriviratNigel S Scrutton
Mar 3, 2020·Metabolic Engineering·Lin-Ping YuGuo-Qiang Chen
Mar 10, 2021·Nature Communications·Xiao-Ran JiangGuo-Qiang Chen
Apr 23, 2021·Essays in Biochemistry·Jian-Wen Ye, Guo-Qiang Chen
Jun 26, 2021·Microbial Biotechnology·Lisa Tietze, Rahmi Lale
Jul 20, 2018·ACS Synthetic Biology·Rui ShenGuo-Qiang Chen

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