Promoter Engineering for Enhanced P(3HB- co-4HB) Production by Halomonas bluephagenesis

ACS Synthetic Biology
Rui ShenGuo-Qiang Chen

Abstract

Promoters for the expression of heterologous genes in Halomonas bluephagenesis are quite limited, and many heterologous promoters function abnormally in this strain. Pporin, a promoter of the strongest expressed protein porin in H. bluephagenesis, is one of the few promoters available for heterologous expression in H. bluephagenesis, yet it has a fixed transcriptional activity that cannot be tuned. A stable promoter library with a wide range of activities is urgently needed. This study reports an approach to construct a promoter library based on the Pporin core region, namely, from the -35 box to the transcription start site, a spacer and an insulator. Saturation mutagenesis was conducted inside the promoter core region to significantly increase the diversity within the promoter library. The promoter library worked in both E. coli and H. bluephagenesis, covering a wide range of relative transcriptional strengths from 40 to 140 000. The library is therefore suitable for the transcription of many different heterologous genes, serving as a platform for protein expression and fine-tuned metabolic engineering of H. bluephagenesis TD01 and its derivative strains. H. bluephagenesis strains harboring the orfZ gene encoding 4HB-CoA tran...Continue Reading

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Citations

Apr 17, 2020·Biotechnology Journal·Ying ZhuChun Li
Jan 4, 2020·The Journal of Microbiology·Jun RenDokyun Na
Nov 18, 2020·Applied Microbiology and Biotechnology·Wen-Hai XieLi-Juan Wang
Dec 11, 2020·Metabolic Engineering·Rosanna YoungPaul S Freemont
Feb 13, 2021·Metabolic Engineering·Yina LinGuo-Qiang Chen
Dec 3, 2020·Synthetic Biology·Duangthip TrisriviratNigel S Scrutton
Mar 3, 2020·Metabolic Engineering·Lin-Ping YuGuo-Qiang Chen
Mar 18, 2021·Critical Reviews in Biotechnology·Kai-Hee HuongA A Amirul
Apr 23, 2021·Essays in Biochemistry·Jian-Wen Ye, Guo-Qiang Chen
Jul 30, 2021·Advanced Materials·Mengyi LiGuo-Qiang Chen
Jul 7, 2021·Journal of Microbiology and Biotechnology·Cai-Yun WangYi-Xuan Zhang

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