Bacilysin overproduction in Bacillus amyloliquefaciens FZB42 markerless derivative strains FZBREP and FZBSPA enhances antibacterial activity

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• 作者：Liming Wu ; Huijun Wu ; Lina Chen ; Ling Lin…
• 关键词：Bacillus amyloliquefaciens ; Bacilysin ; Promoter replacement ; Markerless ; Antibiotic overproduction ; Antibacterial
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2015
• 出版时间：May 2015
• 年：2015
• 卷：99
• 期：10
• 页码：4255-4263
• 全文大小：1,745 KB
• 参考文献：Chen XH, Vater J, Piel J, Franke P, Scholz R, Schneider K, Koumoutsi A, Hitzeroth G, Grammel N, Strittmatter AW, Gottschalk G, Süssmuth RD, Borriss R (2006) Structural and functional characterization of three polyketide synthase gene clusters in Bacillus amyloliquefaciens FZB42. J Bacteriol 188(11):4024-036View Article PubMed Central PubMed
  Chen XH, Koumoutsi A, Scholz R, Eisenreich A, Schneider K, Heinemeyer I, Morgenstern B, Voss B, Hess WR, Reva O, Junge H, Voigt B, Jungblut PR, Vater J, Süssmuth R, Liesegang H, Strittmatter A, Gottschalk G, Borriss R (2007) Comparative analysis of the complete genome sequence of the plant growth-promoting bacterium Bacillus amyloliquefaciens FZB42. Nat Biotechnol 25(9):1007-014View Article PubMed
  Chen XH, Koumoutsi A, Scholz R, Schneider K, Vater J, Süssmuth R, Piel J, Borriss R (2009a) Genome analysis of Bacillus amyloliquefaciens FZB42 reveals its potential for biocontrol of plant pathogens. J Biotechnol 140(1-):27-7View Article PubMed
  Chen XH, Scholz R, Borriss M, Junge H, M?gel G, Kunz S, Borriss R (2009b) Difficidin and bacilysin produced by plant-associated Bacillus amyloliquefaciens are efficient in controlling fire blight disease. J Biotechnol 140(1-):38-4View Article PubMed
  Chmara H, Z?hner H, Borowski E, Milewski S (1984) Inhibition of glucosamine-6-phosphate synthetase from bacteria by anticapsin. J Antibiot 37(6):652-58View Article PubMed
  Claessen D, Emmins R, Hamoen LW, Daniel RA, Errington J, Edwards DH (2008) Control of the cell elongation-division cycle by shuttling of PBP1 protein in Bacillus subtilis. Mol Microbiol 68(4):1029-046View Article PubMed
  Foster JW, Woodruff HB (1946) Bacillin, a new antibiotic substance from a soil isolate of Bacillus subtilis. J Bacteriol 51(3):363-69PubMed Central
  Hilton MD, Alaeddinoglu NG, Demain AL (1988) Bacillus subtilis mutant deficient in the ability to produce the dipeptide antibiotic bacilysin: isolation and mapping of the mutation. J Bacteriol 170(2):1018-020PubMed Central PubMed
  Hoflack L, Seurinck J, Mahillon J (1997) Nucleotide sequence and characterization of the cryptic Bacillus thuringiensis plasmid pGI3 reveal a new family of rolling circle replicons. J Bacteriol 179(16):5000-008PubMed Central PubMed
  Inaoka T, Ochi K (2011) Scandium stimulates the production of amylase and bacilysin in Bacillus subtilis. Appl Environ Microbiol 77(22):8181-183View Article PubMed Central PubMed
  Kenig M, Abraham EP (1976) Antimicrobial activities and antagonists of bacilysin and anticapsin. J Gen Microbiol 94(1):37-5View Article PubMed
  Kenig M, Vandamme E, Abraham EP (1976) The mode of action of bacilysin and anticapsin and biochemical properties of bacilysin-resistant mutants. J Gen Microbiol 94(1):46-4View Article PubMed
  Koumoutsi A, Chen XH, Henne A, Liesegang H, Hitzeroth G, Franke P, Vater J, Borriss R (2004) Structural and functional characterization of gene clusters directing nonribosomal synthesis of bioactive cyclic lipopeptides in Bacillus amyloliquefaciens strain FZB42. J Bacteriol 186(4):1084-096View Article PubMed Central PubMed
  Lambert JM, Bongers RS, Kleerebezem M (2007) Cre-lox-based system for multiple gene deletions and selectable-marker removal in Lactobacillus plantarum. Appl Environ Microbiol 73(4):1126-135View Article PubMed Central PubMed
  Leclère V, Béchet M, Adam A, Guez JS, Wathelet B, Ongena M, Thonart P, Gancel F, Chollet-Imbert M, Jacques P (2005) Mycosubtilin overproduction by Bacillus subtilis BBG100 enhances the organism’s antagonistic and biocontrol activities. Appl Environ Microbiol 71(8):4577-584View Article PubMed Central PubMed
  Leibig M, Krismer B, Kolb M, Friede A, G?tz F, Bertram R (2008) Marker removal in Staphylococci via Cre recombinase and different lox sites. Appl Environ Microbiol 74(5):1316-323View Article PubMed Central PubMed
  Ozcengiz G, Alaeddinoglu NG, Demain AL (1990) Regulation of biosynthesis of bacilysin by Bacillus subtilis. J Ind Microbiol 6(2):91-00View Article PubMed
  Parker JB, Walsh CT (2013) Action and timing of BacC and BacD in the late stages of biosynthesis of the dipeptide antibiotic bacilysin. Biochemistry 52(5):889-01View Article PubMed Central PubMed
  Pomerantsev AP, Sitaraman R, Galloway CR, Kivovich V, Leppla SH (2006) Genome engineering in Bacillus anthracis using Cre recombinase. Infect Immun 74(1):682-93View Article PubMed Central PubMed
  Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
  Scholz R, Molohon KJ, Nachtigall J, Vater J, Markley AL, Süssmuth RD, Mitchell DA, Borriss R (2011) Plantazolicin, a novel microcin B17/streptolysin S-like natural product from Bacillus amyloliquefaciens FZB42. J Bacteriol 193(1):215-24View Article PubMed Central PubMed
  Spizizen J (1958) Transformation of biochemically deficient strains of Bacillus subtilis by deoxyribonucleate. Proc Natl Acad Sci
• 作者单位：Liming Wu (1) (2)
  Huijun Wu (1) (2)
  Lina Chen (1) (2)
  Ling Lin (1) (2)
  Rainer Borriss (3)
  Xuewen Gao (1) (2)
  
  1. Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing, China
  2. Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Education, Nanjing, China
  3. ABiTEP GmbH, Berlin, Germany
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0614

文摘

Bacillus amyloliquefaciens strains FZBREP and FZBSPA were derived from the wild-type FZB42 by replacement of the native bacilysin operon promoter with constitutive promoters P _repB and P _spac from plasmids pMK3 and pLOSS, respectively. These strains contained two antibiotic resistance genes, and markerless strains were constructed by deleting the chloramphenicol resistance cassette and promoter region bordered by two lox sites (lox71 and lox66) using Cre recombinase expressed from the temperature-sensitive vector pLOSS-cre. The vector-encoded spectinomycin resistance gene was removed by high temperature (50?°C) treatment. RT-PCR and qRT-PCR results indicated that P _repB and especially P _spac significantly increased expression of the bac operon, and FZBREP and FZBSPA strains produced up to 170.4 and 315.6?% more bacilysin than wild type, respectively. Bacilysin overproduction was accompanied by enhancement of the antagonistic activities against Staphylococcus aureus (an indicator of bacilysin) and Clavibacter michiganense subsp. sepedonicum (the causative agent of potato ring rot). Both the size and degree of ring rot-associated necrotic tubers were decreased compared with the wild-type strain, which confirmed the protective effects and biocontrol potential of these genetically engineered strains.