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Function of degQ and sfp and Their Effects on Fengycin Productivity of Bacillus subtilis

LI Xiang, WU Huijun, ZHANG Yang, LU Qinghui, GAO Xuewen   

  1. Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education/College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2014-08-30 Revised:1900-01-01 Online:2015-04-08 Published:2015-04-08

Abstract: Fengycin, a lipopeptide synthesized non-ribosomally by Bacillus spp., plays an important role in controlling plant fungus diseases. The sfp gene, which encodes 4'-phosphopantetheine transferase, is required for all cells of Bacillus subtilis to become producers of the lipopeptide. The degQ is a pleiotropic gene affecting the expression of a number of secreted gene products. For studying the effect and function of degQ and sfp activation on fengycin productivity of Bacillus subtilis 168, we cloned the sfp gene from Bacillus amyloliquefaciens FZB42 and the degQ gene from Bacillus subtilis 168, constructed expression vectors pP43-degQ, pMK3-sfp andpP43-degQ-sfp, and gained three genetically engineered bacterial strains by transformation of the strain 168 which was a nonfengycin-producer. Antimicrobial activities experiment showed that both the living cell and its lipopeptide extract from 168-degQ-sfp inhibited the growth of Sclerotinia sclerotiorum on plates, whereas 168, 168-degQ and 168-sfp did not. MALDI-TOF-MS analysis demonstrated that only 168-degQ-sfp could produce fengycin. Real-time PCR result showed that fengycin synthase gene cluster ppsABCDE in 168-degQ-sfp were expressed 6—16 folds higher than that in wild-type, while not in 168-degQ and 168-sfp were. This study indicated that the two functional genes degQ and sfp could jointly enhance the expression of fengycin synthase gene cluster, and increase fengycin production.

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