Characterization and cloning of the genes encoding enterocin 1071A and enterocin 1071B, two antimicrobial peptides produced by Enterococcus faecalis BFE 1071

被引:97
作者
Balla, E
Dicks, LMT [1 ]
Du Toit, M
van der Merwe, MJ
Holzapfel, WH
机构
[1] Univ Stellenbosch, Dept Microbiol, ZA-7600 Stellenbosch, South Africa
[2] Univ Stellenbosch, Dept Biochem, ZA-7600 Stellenbosch, South Africa
[3] Fed Res Ctr Nutr, Inst Hyg & Toxicol, D-76131 Karlsruhe, Germany
关键词
D O I
10.1128/AEM.66.4.1298-1304.2000
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The pH-neutral cell supernatant of Enterococcus faccalis BFE 1071, isolated from the feces of minipigs in Gottingen, inhibited the growth of Enterococcus spp, and a few other gram-positive bacteria. Ammonium sulfate precipitation and cation-exchange chromatography of the cell supernatant, followed by mass spectrometry analysis, yielded two bacteriocin-like peptides of similar molecular mass: enterocin 1071A (4.285 kDa) and enterocin 1071B (3.899 kDa). Both peptides are always isolated together. The peptides are heat resistant (100 degrees C, 60 min; 50% of activity remained after 15 min at 121 degrees C), remain active after 30 min of incubation at pH 3 to 12, and are sensitive to treatment with proteolytic enzymes. Curing experiments indicated that the genes encoding enterocins 1071A and 1071B are located on a 50-kbp plasmid (pEF1071). Conjugation of plasmid pEF1071 to E.faecalis strains FA2-2 and OGX1 resulted in the expression of two active peptides with sizes identical to those of enterocins 1071A and 1071B, Sequencing of a DNA insert of 9 to 10 kbp revealed two open reading Frames, ent1071A and ent1071B, which coded for 39- and M-amino-acid peptides? respectively. The deduced amino acid sequence of the mature Ent1071A and Ent1071B peptides showed 64 and 61% homology with the alpha and beta peptides of lactococcin G, respectively. This is the first report of tno nem antimicrobial peptides representative of a fourth type of E. faecalis bacteriocin.
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页码:1298 / 1304
页数:7
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