Specificity and Application of the Lantibiotic Protease NisP
Lantibiotics are ribosomally produced and posttranslationally modified peptides containing several lanthionine residues. They exhibit substantial antimicrobial activity against Gram-positive bacteria, including relevant pathogens. The production of the model lantibiotic nisin minimally requires the...
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2018-02-01
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doaj-26a83ce0b5864c2e90a8c0ab36e37a342020-11-25T00:56:34ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-02-01910.3389/fmicb.2018.00160329097Specificity and Application of the Lantibiotic Protease NisPManuel Montalbán-LópezJingjing DengAuke J. van HeelOscar P. KuipersLantibiotics are ribosomally produced and posttranslationally modified peptides containing several lanthionine residues. They exhibit substantial antimicrobial activity against Gram-positive bacteria, including relevant pathogens. The production of the model lantibiotic nisin minimally requires the expression of the modification and export machinery. The last step during nisin maturation is the cleavage of the leader peptide. This liberates the active compound and is catalyzed by the cell wall-anchored protease NisP. Here, we report the production and purification of a soluble variant of NisP. This has enabled us to study its specificity and test its suitability for biotechnological applications. The ability of soluble NisP to cleave leaders from various substrates was tested with two sets of nisin variants. The first set was designed to investigate the influence of amino acid variations in the leader peptide or variations around the cleavage site. The second set was designed to study the influence of the lanthionine ring topology on the proteolytic efficiency. We show that the substrate promiscuity is higher than has previously been suggested. Our results demonstrate the importance of the arginine residue at the end of the leader peptide and the importance of lanthionine rings in the substrate for specific cleavage. Collectively, these data indicate that NisP is a suitable protease for the activation of diverse heterologously expressed lantibiotics, which is required to release active antimicrobial compounds.http://journal.frontiersin.org/article/10.3389/fmicb.2018.00160/fulllantibioticnisinbacteriocinLactococcus lactisNisPleader peptidase |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Manuel Montalbán-López Jingjing Deng Auke J. van Heel Oscar P. Kuipers |
spellingShingle |
Manuel Montalbán-López Jingjing Deng Auke J. van Heel Oscar P. Kuipers Specificity and Application of the Lantibiotic Protease NisP Frontiers in Microbiology lantibiotic nisin bacteriocin Lactococcus lactis NisP leader peptidase |
author_facet |
Manuel Montalbán-López Jingjing Deng Auke J. van Heel Oscar P. Kuipers |
author_sort |
Manuel Montalbán-López |
title |
Specificity and Application of the Lantibiotic Protease NisP |
title_short |
Specificity and Application of the Lantibiotic Protease NisP |
title_full |
Specificity and Application of the Lantibiotic Protease NisP |
title_fullStr |
Specificity and Application of the Lantibiotic Protease NisP |
title_full_unstemmed |
Specificity and Application of the Lantibiotic Protease NisP |
title_sort |
specificity and application of the lantibiotic protease nisp |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Microbiology |
issn |
1664-302X |
publishDate |
2018-02-01 |
description |
Lantibiotics are ribosomally produced and posttranslationally modified peptides containing several lanthionine residues. They exhibit substantial antimicrobial activity against Gram-positive bacteria, including relevant pathogens. The production of the model lantibiotic nisin minimally requires the expression of the modification and export machinery. The last step during nisin maturation is the cleavage of the leader peptide. This liberates the active compound and is catalyzed by the cell wall-anchored protease NisP. Here, we report the production and purification of a soluble variant of NisP. This has enabled us to study its specificity and test its suitability for biotechnological applications. The ability of soluble NisP to cleave leaders from various substrates was tested with two sets of nisin variants. The first set was designed to investigate the influence of amino acid variations in the leader peptide or variations around the cleavage site. The second set was designed to study the influence of the lanthionine ring topology on the proteolytic efficiency. We show that the substrate promiscuity is higher than has previously been suggested. Our results demonstrate the importance of the arginine residue at the end of the leader peptide and the importance of lanthionine rings in the substrate for specific cleavage. Collectively, these data indicate that NisP is a suitable protease for the activation of diverse heterologously expressed lantibiotics, which is required to release active antimicrobial compounds. |
topic |
lantibiotic nisin bacteriocin Lactococcus lactis NisP leader peptidase |
url |
http://journal.frontiersin.org/article/10.3389/fmicb.2018.00160/full |
work_keys_str_mv |
AT manuelmontalbanlopez specificityandapplicationofthelantibioticproteasenisp AT jingjingdeng specificityandapplicationofthelantibioticproteasenisp AT aukejvanheel specificityandapplicationofthelantibioticproteasenisp AT oscarpkuipers specificityandapplicationofthelantibioticproteasenisp |
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1725226595037216768 |