Enterolysin A, a Cell Wall-Degrading Bacteriocin fromEnterococcus faecalisLMG 2333

Applied and Environmental Microbiology - Tập 69 Số 5 - Trang 2975-2984 - 2003
Trine Nilsen1, Ingolf F. Nes1, Helge Holo1
1Laboratory of Microbial Gene Technology, Agricultural University of Norway, N-1432, As, Norway

Tóm tắt

ABSTRACTA novel antimicrobial protein, designated enterolysin A, was purified from anEnterococcus faecalisLMG 2333 culture. Enterolysin A inhibits growth of selected enterococci, pediococci, lactococci, and lactobacilli. Antimicrobial activity was initially detected only on solid media, but by growing the bacteria in a fermentor under optimized production conditions (MRS broth with 4% [wt/vol] glucose, pH 6.5, and a temperature between 25 and 35°C), the bacteriocin activity was increased to 5,120 bacteriocin units ml−1. Enterolysin A production was regulated by pH, and activity was first detected in the transition between the logarithmic and stationary growth phases. Killing of sensitive bacteria by enterolysin A showed a dose-response behavior, and the bacteriocin has a bacteriolytic mode of action. Enterolysin A was purified, and the primary structure was determined by combined amino acid and DNA sequencing. This bacteriocin is translated as a 343-amino-acid preprotein with ansec-dependent signal peptide of 27 amino acids, which is followed by a sequence corresponding to the N-terminal part of the purified protein. Mature enterolysin A consists of 316 amino acids and has a calculated molecular weight of 34,501, and the theoretical pI is 9.24. The N terminus of enterolysin A is homologous to the catalytic domains of different cell wall-degrading proteins with modular structures. These include lysostaphin, ALE-1, zoocin A, and LytM, which are all endopeptidases belonging to the M37 protease family. The N-terminal part of enterolysin A is linked by a threonine-proline-rich region to a putative C-terminal recognition domain, which shows significant sequence identity to two bacteriophage lysins.

Từ khóa


Tài liệu tham khảo

10.1093/nar/25.17.3389

EMBO J. 1996 15

10.1128/aem.45.6.1808-1815.1983

10.1128/AEM.67.9.3888-3896.2001

10.1128/AEM.66.1.23-28.2000

10.1128/aem.57.4.1265-1267.1991

10.1016/S0021-9258(19)70112-4

Cai Y. and Y. Benno. 16 March 1998 posting date. Determination of 16S ribosomal DNA sequences of Enterococcus faecalis . [Online.] http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=nucleotide&list_uids=298272&dopt=GenBank .

Casaus, P., T. Nilsen, L. M. Cintas, I. F. Nes, P. E. Hernandez, and H. Holo. 1998. Enterocin B, a new bacteriocin from Enterococcus faecium T136 which can act synergistically with enterocin A. Microbiology143:2287-2294.

10.1016/S0021-9258(19)47165-2

10.1128/jb.179.2.548-551.1997

10.1128/aem.61.7.2643-2648.1995

Cooper, V. J. C., and G. P. C. Salmond. 1993. Molecular analysis of the major cellulase (CelV) of Erwinia carotovora: evidence for an evolutionary ‘mix and match’ of enzyme domains. Mol. Gen. Genet.241:342-350.

10.1016/0006-291X(88)90188-X

De Vuyst L. and E. J. Vandamme. 1994. Antimicrobial potential of lactic acid bacteria p. 91-142. In L. De Vuyst and E. J. Vandamme (ed.) Bacteriocins of lactic acid bacteria 1st ed. Blackie Academic & Professional London United Kingdom.

10.1099/00221287-142-4-817

10.1111/j.1365-2958.1995.mmi_18040631.x

10.1128/jb.178.8.2232-2237.1996

10.1139/m97-151

10.1128/aem.45.1.205-211.1983

10.1128/jb.178.22.6608-6617.1996

10.1016/S0006-291X(89)80117-2

10.1007/BF00331163

10.1128/jb.173.12.3879-3887.1991

10.1128/mr.59.2.171-200.1995

J. Bacteriol. 1990 172

Arch. Virol. 2000 145

10.1016/S0021-9258(18)53203-8

10.1038/227680a0

10.1111/j.1365-2958.1995.tb02331.x

10.1139/m89-125

10.1128/aem.61.1.175-179.1995

10.1007/BF00395929

10.1093/protein/10.1.1

10.1128/JB.180.7.1848-1854.1998

Patel R. K. E. Piper M. S. Rouse J. M. Steckelberg J. R. Uhl P. Kohner M. K. Hopkins F. R. Cockerill and B. C. Kline. 29 December 1997 posting date. Determination of 16S ribosomal DNA sequences of Enterococcus faecium Enterococcus faecalis Enterococcus gallinarum and Enterococcus casseliflavus . [Online.] http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=nucleotide&list_uids=2828135&dopt=GenBank .

Pot B. P. Vandamme and K. Kersters. 1994. Analysis of electrophoretic whole-organism protein fingerprints p. 493-521. In M. Goodfellow and A. O'Donnell (ed.) Chemical methods in prokaryotic systematics. John Wiley & Sons Chichester United Kingdom.

10.1007/BF00337717

10.1128/jb.179.11.3625-3631.1997

10.1099/13500872-145-4-801

10.1073/pnas.84.5.1127

10.1016/0378-1119(83)90168-3

Riley, M. A. 1993. Molecular mechanisms of colicin evolution. Mol. Biol. Evol.10:1380-1395.

10.1128/aem.55.8.1901-1906.1989

Schindler C. A. and V. T. Schuhardt. 1964. Lysostaphin: a new bacteriolytic agent for the staphylococcus. Proc. Natl. Acad. Sci. USA 51: 414-421.

10.1128/aem.62.12.4536-4541.1996

10.1016/S0378-1119(96)00859-1

10.1128/jb.179.4.1193-1202.1997

10.1016/S0723-2020(11)80415-7

10.1128/aem.62.4.1283-1286.1996

10.1128/jb.155.2.866-871.1983

10.1016/0378-1119(91)90547-O

Thông tin
Thông tin xuất bản

Applied and Environmental Microbiology

Tập 69 Số 5

2975-2984

Thông tin tác giả