title: The novel Shewanella putrefaciens-infecting bacteriophage Spp001: genome sequence and lytic enzymes.
abstract: Shewanella putrefaciens has been identified as a specific spoilage organism commonly found in chilled fresh fish, which contributes to the spoilage of fish products. Limiting S. putrefaciens growth can extend the shelf-life of chilled fish. Endolysins, which are lytic enzymes produced by bacteriophages, have been considered an alternative to control bacterial growth, and have been useful in various applications, including food preservation. We report here, for the first time, the complete genome sequence of a novel phage Spp001, which lyses S. putrefaciens Sp225. The Spp001 genome comprises a 54,789-bp DNA molecule with 67 open reading frames and an average total G + C content of 49.42 %. In silico analysis revealed that the Spp001 open reading frames encode various putative functional proteins, including an endolysin (ORF 62); however, no sequence for genes encoding the holin polypeptides, which work in concert with endolysins, was identified. To examine further the lytic activity of Spp001, we analyzed the lytic enzyme-containing fraction from phages released at the end of the phage lytic cycle in S. putrefaciens, using diffusion and turbidimetric assays. The results show that the partially purified extract contained endolysin, as indicated by a high hydrolytic activity towards bacterial peptidoglycan decrease in the OD590 value by 0.160 in 15 min. The results will allow further investigation of the purification of natural Spp001 endolysin, the extension of Spp001 host range, and the applications of the phage-encoded enzymes.
authors.affiliation: Food Safety Laboratory, Ocean University of China, Qingdao, 266003, People's Republic of China.
authors.forename: Feng
authors.initials: F
authors.lastname: Han
authors.forename: Meng
authors.initials: M
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Lin
authors.forename: Jingxue
authors.initials: J
authors.lastname: Wang
authors.forename: Limin
authors.initials: L
authors.lastname: Cao
authors.forename: Muhammad Naseem
authors.initials: MN
authors.lastname: Khan
mesh_terms: D000818:Animals; D001435:Bacteriophages / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D004279:DNA, Viral / Q000737:chemistry; D010450:Endopeptidases / Q000235:genetics; D005814:Genes, Viral; D016679:Genome, Viral; D016366:Open Reading Frames; D020614:Shewanella putrefaciens / Q000821:virology; D014764:Viral Proteins / Q000235:genetics
chemical_list: D004279:DNA, Viral; D014764:Viral Proteins; D010450:Endopeptidases; C001143:endolysin
references.citation: J Bacteriol. 2004 Jul;186(14):4808-12
references.pmid: 15231813
references.citation: Gene. 2012 Oct 10;507(2):125-34
references.pmid: 22868206
references.citation: Trends Microbiol. 2005 Oct;13(10):491-6
references.pmid: 16125935
references.citation: Int J Food Microbiol. 2003 Jul 25;84(2):225-35
references.pmid: 12781945
references.citation: J Ind Microbiol Biotechnol. 2013 Oct;40(10):1105-16
references.pmid: 23907252
references.citation: Curr Microbiol. 2009 Jun;58(6):609-15
references.pmid: 19267155
references.citation: Curr Opin Biotechnol. 2011 Apr;22(2):164-71
references.pmid: 21093250
references.citation: BMC Microbiol. 2012 Dec 19;12:297
references.pmid: 23249212
references.citation: Annu Rev Food Sci Technol. 2010;1:449-68
references.pmid: 22129344
references.citation: Mol Microbiol. 1995 Jun;16(6):1231-41
references.pmid: 8577256
references.citation: J Bacteriol. 1993 Jul;175(14):4550-3
references.pmid: 8331084
references.citation: PLoS One. 2012;7(4):e34585
references.pmid: 22514640
references.citation: Anal Biochem. 2001 Apr 1;291(1):149-54
references.pmid: 11262168
references.citation: J Clin Microbiol. 2003 May;41(5):2240-1
references.pmid: 12734291
references.citation: Appl Environ Microbiol. 2000 Jul;66(7):2951-8
references.pmid: 10877791
references.citation: Proc Natl Acad Sci U S A. 2001 Mar 27;98(7):4107-12
references.pmid: 11259652
references.citation: Int J Food Microbiol. 1996 Nov;33(1):121-37
references.pmid: 8913813
references.citation: Bioeng Bugs. 2010 Jan-Feb;1(1):9-16
references.pmid: 21327123
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: J Bacteriol. 1981 Jan;145(1):661-3
references.pmid: 7462154
references.citation: Methods Mol Biol. 2009;501:69-76
references.pmid: 19066811
references.citation: Appl Environ Microbiol. 1995 Sep;61(9):3391-9
references.pmid: 16535125
references.citation: Vet Microbiol. 2008 Jul 27;130(1-2):107-17
references.pmid: 18242012
references.citation: Appl Environ Microbiol. 2013 Mar;79(6):1956-68
references.pmid: 23335772
references.citation: J Exp Med. 1971 May 1;133(5):1105-17
references.pmid: 4928818
references.citation: J Microbiol Immunol Infect. 2012 Apr;45(2):151-3
references.pmid: 22154677
references.citation: BMC Microbiol. 2012 Mar 15;12:33
references.pmid: 22416675
references.citation: Gene. 2002 Oct 16;299(1-2):227-34
references.pmid: 12459270
references.citation: PLoS One. 2012;7(5):e36991
references.pmid: 22615864
references.citation: Aust N Z J Surg. 2000 Nov;70(11):816-7
references.pmid: 11147445
references.citation: J Sci Food Agric. 2014 Jun;94(8):1657-62
references.pmid: 24214837
references.citation: J Appl Bacteriol. 1993 Apr;74(4):444-51
references.pmid: 8486552
references.citation: Mikrobiyol Bul. 2012 Jan;46(1):117-21
references.pmid: 22399180
references.citation: Viruses. 2010 Mar;2(3):710-30
references.pmid: 21994654
references.citation: Appl Environ Microbiol. 2002 Nov;68(11):5311-7
references.pmid: 12406719
references.citation: Mol Microbiol. 2007 Sep;65(5):1334-44
references.pmid: 17697255
references.citation: Clin Infect Dis. 1995 May;20(5):1327-32
references.pmid: 7620019
references.citation: Antimicrob Agents Chemother. 2011 Feb;55(2):738-44
references.pmid: 21098252
references.citation: Res Microbiol. 2012 Apr;163(3):233-41
references.pmid: 22289622
references.citation: Appl Microbiol Biotechnol. 2013 May;97(10):4369-75
references.pmid: 22832988
references.citation: Curr Opin Microbiol. 2005 Aug;8(4):480-7
references.pmid: 15979390