Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (24): 5142-5149.doi: 10.3864/j.issn.0578-1752.2013.24.008

• PLANT PROTECTION • Previous Articles     Next Articles

The Operon, Structure and Biological Activities of the Lipopeptide Fengycin Produced by Bacillus subtilis 916

 LUO  Chu-Ping-1, 2 , WANG  Xiao-Yu-1, ZHOU  Hua-Fei-1, 2 , LIU  You-Zhou-1, CHEN  Zhi-Yi-1, 2   

  1. 1.Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014;
    2.College of Plant Protection, Nanjing Agricultural University, Nanjing 210095)
  • Received:2013-05-13 Online:2013-12-16 Published:2013-07-29

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Abstract: 【Objective】The objective of this study is to clone and sequence the Fen operon responsible for synthesis of the fengycin from Bacillus subtilis 916 and determine the structure and biological activities of the Fen operon. 【Method】Genome sequence and PCR were performed to clone the Fen operon. Analysis of the Fen operon genetic structure was made by using bioinformatics. The Fen mutant BSFG was constructed by the homologous recombination. Fengycin produced by the Bs916 and mutant was analyzed by the reversed-phase high-performance liquid chromatography (HPLC). The molecular weight of the fengycin was determined by the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MS). The antifungal activities and hemolytic activities were evaluated in the form of a flat plate. 【Result】The 37.67 kb Fen operon responsible for synthesis of fengycin was identified and cloned from B. subtilis 916. The operon contained five ORFs designated FenA, FenB, FenC, FenD andFenE, respectively. The amino acid sequences encoded by the Bs916 Fen operon share 65% similarity with the counterpart amino acid sequences from the Bs168 Fen operon. The results of HPLC analysis of lipopeptides produced by wild-type Bs916 and mutant BSFG showed the Fen operon responsible for fengycin synthesis. The molecular weights determined by MS were 1 449.8, 1 463.9, 1 477.9 and 1 491.9, 1 505.9 Da showed fengycin produced by Bs916 contained five homologues. The first three homologues were differed by a structure of –CH2 and their peptide moiety primary structure were [cyclo-([cyclo-(Glu-Orn-Tyr-Thr-Glu-Ala-Pro- Gln-Tyr-β-amino fatty acid)]. The last two homologues were also differed by a structure of -CH2 and their peptide moiety primary structure were Val (6) instead Ala (6) compared to homologues above. Although the mutant BSFG decreased clearly in antifungal activities, its hemolytic activities showed no obvious difference compared to wild type Bs916. 【Conclusion】This paper reported the cloning, sequencing and characterization of a whole operon Fen which is responsible for synthesis of fengycin. Through bio-information and chemical analysis, the authors confirmed that the fengycin produced by the Bs916 contains five homologues. The fengycin synthesized by Fen plays a crucial part in Bs916 bio-control activities.

Key words: Bacillus subtilis , fengycin , operon , structure , biological activity

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