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Improved Prodigiosin Production by Relieving CpxR Temperature-Sensitive Inhibition
Yang Sun1, Lijun Wang1, Xuewei Pan1
1The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.
Frontiers in Bioengineering and Biotechnology
|June 26, 2020
Summary
Researchers found that the CpxR protein represses prodigiosin (PG) production in Serratia marcescens. Deleting cpxR or enhancing precursor pathways significantly boosted PG yields, offering a new strategy for microbial production.
Area of Science:
- Microbiology
- Molecular Biology
- Biotechnology
Background:
- Prodigiosin (PG) is a secondary metabolite produced by Serratia marcescens.
- CpxR is a transcriptional regulator belonging to the OmpR family in Gram-negative bacteria.
Purpose of the Study:
- To investigate the role of CpxR in regulating prodigiosin biosynthesis in Serratia marcescens.
- To develop an enhanced strategy for increasing prodigiosin production through genetic engineering.
Main Methods:
- Insertion mutagenesis of the cpxR gene in Serratia marcescens JNB 5-1 using transposon Tn5G.
- Electrophoretic mobility shift assay (EMSA) to determine CpxR binding to the pig gene promoter.
- Gene expression analysis of the pig gene cluster and precursor biosynthesis pathways.
- Genetic engineering by inserting a fusion segment of proC, serC, and metH genes into the cpxR gene.
Main Results:
- Mutation of cpxR significantly increased PG production.
- CpxR was found to bind to the pig gene promoter, repressing transcription of PG biosynthesis genes.
- Deletion of cpxR led to increased transcription of both the pig gene cluster and genes involved in proline, serine, and methionine precursor synthesis.
- Engineered Serratia marcescens with a fusion segment in the cpxR gene achieved a highest PG titer of 5.83 g/L, a 41.9% increase.
Conclusions:
- CpxR plays a crucial role in repressing prodigiosin biosynthesis in Serratia marcescens.
- Disrupting CpxR function or enhancing precursor pathways presents a viable strategy for increasing PG production.
- This study provides a novel approach for the metabolic engineering of Serratia marcescens to improve prodigiosin yields.
