Analysis of the Genetic Determinant for Production of the Peptide Antibiotic Nisin

Dodd, Helen M.; Horn, Nikki; Gasson, Michael J.

doi:10.1099/00221287-136-3-555

Cited by 236 publications

(165 citation statements)

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“…The first lan-!ibiotic to be characterised was nisin, produced by strains of Lactococcus lactis carrying a transposon containing genes coding for the nisin precursor and for proteins involved in aisin biosynthesis and resistance [2][3][4][5]. Nisin has been quite videly used as a food preservative, notably in cheese and ,~ther dairy products and in canned vegetables, for some 30 .,ears [6,7].…”

Section: ~ Introductionmentioning

confidence: 99%

Structure‐activity relationships in the peptide antibiotic nisin: antibacterial activity of fragments of nisin

Chan¹,

Leyland²,

Clark³

et al. 1996

FEBS Letters

122

160

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The post-translationally modified peptide antibiotic nisin has been cleaved by a number of proteases and the fragments produced purified, characterised chemically, and assayed for activity in inhibiting the growth of Lactococcus lactis MG1614 and Micrococcus luteus NCDO8166. These results provide information on the importance of different parts of the nisin molecule for its growth-inhibition activity. Removal of the C-terminal five residues leads to approximately a 10-fold decrease in potency, while removal of a further nine residues, encompassing two of the lanthionine rings, leads to a 100-fold decrease. There are some differences between analogous fragments of nisin and subtilin, suggesting possible subtle differences in mode of action. Cleavage within, or removal of, lanthionine ring C essentially abolishes the activity of nisin. The fragment ulsin t-12 is inactive itself, and specifically antagonises the growth-inhibitory action of nisin. These results are discussed in lerms of current models for the mechanism of action of nisin.Key words: Nisin; Lantibiotic; Peptide antibiotic; I'roteolysis; Structure-activity relationships mation of voltage-dependent pores in biological membranes [8][9][10], although recent evidence indicates that the inhibition of the outgrowth of spores by nisin and subtilin takes place by a different mechanism [11,12]. We have been studying the structure-activity relationships in nisin and subtilin, by both genetic and chemical modification of the structure [12][13][14][15][16][17][18][19][20], with a view to understanding their mechanism of action and to developing new derivatives with desirable properties. We now report the preparation, characterisation and anti-bacterial activity of a number of proteolytic fragments of nisin and subtilin, which allow us to define the parts of the molecule most important for biological activity. Materials and methodsNisin and subtilin were prepared as previously described [14,15]; [Ser33]-nisin, in which the serine residue at position 33 has escaped processing to a dehydroalanine residue, was isolated as a minor component of commercial nisin. All proteases were obtained from Sigma Chemical Co., Poole, Dorset, UK. ~. IntroductionThe post-translationally modified peptide antibiotics known :ts 'lantibiotics' contain cyclic structures formed by lanthioaine or 3-methyl-lanthionine residues, and often also dehydroalanine and/or dehydrobutyrine residues [1]. The first lan-!ibiotic to be characterised was nisin, produced by strains of Lactococcus lactis carrying a transposon containing genes coding for the nisin precursor and for proteins involved in aisin biosynthesis and resistance [2][3][4][5]. Nisin has been quite videly used as a food preservative, notably in cheese and ,~ther dairy products and in canned vegetables, for some 30 .,ears [6,7]. It inhibits the growth of a wide range of Gram~ositive organisms, and also inhibits the germination and/or mtgrowth of spores of Bacillus and Clostridium species [6]. Fhe growth-inhibitory activity of nisi...

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Section: ~ Introductionmentioning

confidence: 99%

Structure‐activity relationships in the peptide antibiotic nisin: antibacterial activity of fragments of nisin

Chan¹,

Leyland²,

Clark³

et al. 1996

FEBS Letters

122

160

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“…5A), and is distinct from the classAll leader peptides that are found for other lantibiotics such as streptococcin A-FF22 (Hynes et al, 1993), salivaricin A (Ross et al, 1993) and lacticin 481 (Piard et al, 1993). The class-A1 leader peptides may serve several purposes, such as directing the transport of the peptides across the cytoplasmic membrane and acting as a template in the maturation reaction(s) of the (Buchmann et al, 1988;Dodd et al, 1990), nisin Z (Mulders et al, 1991), subtilin , epidermin (Schnell et al, 1988), gallidermin (Schnell et al, 1989) and Pep5 . Gaps are indicated by hyphens.…”

Section: Resultsmentioning

confidence: 99%

“…Knowledge of the gene sequence moreover offers insight into the biosynthesis of the mature protein. For a number of lantibiotics, the structural genes have been cloned and sequenced Buchmann et al, 1988;Schnell et al, 1988Schnell et al, , 1989Kaletta et al, , 1991Dodd et al, 1990;Mulders et al, 1991;Hynes et al, 1993;Piard et al, 1993;Ross et al, 1993). These sequences reveal that lantibiotics are synthesized as preproteins, consisting of an N-terminal leader peptide, and a C-terminal propeptide, which undergoes maturation to yield the fully modified protein.…”

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confidence: 99%

Elucidation of the Primary Structure of the Lantibiotic Epilancin K7 from Staphylococcus epidermidis K7. Cloning and Characterisation of the Epilancin-K7-Encoding Gene and NMR Analysis of Mature Epilancin K7

Kamp¹,

Hooven²,

Konings³

et al. 1995

Eur J Biochem

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Elucidation of the primary structure of the lantibiotic epilancin K7 from Staphylococcus epidermidis K7. Cloning and characterisation of the epilancin-K7-encoding gene and NMR analysis of mature epilancin K7 Kamp, Mart van de; Hooven, Henno W. van den; Konings, Ruud N.H.; Bierbaum, Gabriele; Sahl, Hans-Georg; Kuipers, Oscar; Siezen, Roland J.; Vos, Willem M. de; Hilbers, Cornelis W.; Ven, Frank J.M. van de Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons the number of authors shown on this cover page is limited to 10 maximum. Download date: 10-05-2018Eur. J. Biochem. 230, 587-600 (1995) Lantibiotics are bacteriocins that contain unusual amino acids such as lanthionines and a$-didehydro residues generated by posttranslational modification of a ribosomally synthesized precursor protein. The structural gene encoding the novel lantibiotic epilancin K7 from Staphylococcus epidemzidis K7 was cloned and its nucleotide sequence was determined. The gene, which was named elkA, codes for a 55-residue preprotein, consisting of an N-terminal 24-residue leader peptide, and a C-terminal 31 -residue propeptide which is posttranslationally modified and processed to yield mature epilancin K7. In common with the type-A lantibiotics nisin A and nisin Z , subtilin, epidermin, gallidermin and Peps, pre-epilancin K7 has a so-called class-A1 leader peptide. Downstream and upstream of the elk4 gene, the starts of two open-reading-frames, named elkP and elkT, were identified. The elkP and elkT genes presumably encode a leader peptidase and a translocator protein, respectively, which may be involved in the processing and export of epilancin K7. The amino acid sequence of the unmodified pro-epilancin K7, deduced from the elkA gene sequence, is in full agreement with the amino acid sequence of mature epilancin K7, determined previously by means of NMR spectroscopy Biochern. 227, 757-7711. The first residue of mature epilancin K7 appears to be modified in a way that has not been described for any other lantibiotic so far. NMR experiments show that the elkA-encoded serine residue at position + I of pro-epilancin K7 is modified to a 2-hydroxypropionyl residue in the mature protein.

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“…lactis subsp. lactis LL27 was nisin, PCR analysis was performed using the published sequences of the nisin structural gene (Dodd et al 1990). Two primers complementary to the sequences occurring proximal to 5' and 3' ends of the nisin A structural gene were used to amplify the nisin gene from the genomic DNA of LL27 and Lc.…”

Section: Characterisation Of Bacteriocinmentioning

confidence: 99%

Identification and characterisation of antimicrobial activity of nisin a produced by Lactococcus lactis subsp. lactis LL27

Şanlibaba¹,

Akkoç²,

Akçelik³

2009

Czech J. Food Sci.

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AbstractŞanlibaba P., Akkoç N., Akçelik M. (2009): Identification and characterisation of antimicrobial activity of nisin A produced by Lactococcus lactis subsp. lactis LL27. Czech J. Food Sci., 27: 55-64.In this study, bacteriocin producing lactococcal strains were isolated from Turkish raw milk samples. Among these isolates, LL27 had the highest inhibition activity against Gram-positive indicator strains, and was selected for further analyses. DNA sequencing of 16S ribosomal RNA gene demonstrated that the isolate was Lactococcus lactis subsp. lactis. The gene encoding the bacteriocin in this strain was found to be identical to that of nisin A using direct PCR sequence methods. The bacteriocin was completely inactivated by α-chymotrypsin and proteinase K and partially inactivated by lipase and α-amylase. pH and heat stability characteristics were found to be identical with those of the control nisin. The inhibitory activity of the bacteriocin produced by LL27 was also evaluated against Gram-negative bacteria in combination with heat and freezing treatments. The results obviously showed that the production level of nisin of the Lc. lactis LL27 had a significant inhibitory effect on the pathogenic Gram-negative strains with the heating and freezing processes which are commonly used in the food processing.

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Analysis of the Genetic Determinant for Production of the Peptide Antibiotic Nisin

Cited by 236 publications

References 49 publications

Structure‐activity relationships in the peptide antibiotic nisin: antibacterial activity of fragments of nisin

Structure‐activity relationships in the peptide antibiotic nisin: antibacterial activity of fragments of nisin

Elucidation of the Primary Structure of the Lantibiotic Epilancin K7 from Staphylococcus epidermidis K7. Cloning and Characterisation of the Epilancin-K7-Encoding Gene and NMR Analysis of Mature Epilancin K7

Identification and characterisation of antimicrobial activity of nisin a produced by Lactococcus lactis subsp. lactis LL27

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