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Improvement of pyoluteorin production in Pseudomonas protegens H78 through engineering its biosynthetic and regulatory pathways

  • Huimin Shi
  • Xianqing HuangEmail author
  • Zheng Wang
  • Yejun Guan
  • Xuehong Zhang
Applied genetics and molecular biotechnology

Abstract

Pyoluteorin (Plt) is a PKS-NRPS hybrid antibiotic that is produced by Pseudomonas spp. and shows strong antifungal and antibacterial activities. Pseudomonas protegens H78, which was isolated from the rape rhizosphere in Shanghai, can produce a large array of secondary metabolites, including antibiotics and siderophores. Plt is produced at low levels in the H78 wild-type strain. This study aimed to improve Plt production through combinatory genetic engineered strategies. Plt production was significantly enhanced (by14.3-fold) in the strain engineered by the following steps: (1) deletion of the translational repressor gene rsmE in the Gac/Rsm-RsmE pathway; (2) deletion of the ATP-dependent protease gene lon that encodes a potential enzyme that degrades positive regulators; (3) deletion of the negative regulatory gene pltZ of the Plt ABC-type transporter operon pltIJKNOP; (4) deletion of an inhibitory sequence within the operator of the transcriptional activator gene pltR; and (5) overexpression of the pltIJKNOP transport operon. The Plt production of the final engineered strain was increased to 214 from 15 μg ml−1 in the H78 wild-type strain. In addition, the pltA gene in the pltLABCDEFG biosynthetic operon was characterized as the gene encoding the rate-limiting enzyme in the Plt biosynthetic pathway of H78. However, overexpression of the rate-limiting enzyme gene pltA or the transcriptional activator gene pltR did not further improve Plt biosynthesis in the above multiple-gene knockout strains.

Keywords

Plant growth-promoting rhizobacteria Pseudomonas Antibiotic biosynthesis Genetic engineering Metabolic engineering 

Notes

Funding information

This study was supported by the National Natural Science Foundation of China (31470196 and 31270083 to Xianqing Huang).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants by any of the authors.

Supplementary material

253_2019_9732_MOESM1_ESM.pdf (761 kb)
ESM 1 (PDF 761 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Microbial Metabolism, Joint International Laboratory on Metabolic & Developmental Sciences, School of Life Sciences & BiotechnologyShanghai Jiao Tong UniversityShanghaiChina

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