Bioengineering nisin to overcome the nisin resistance protein

Field, Des; Blake, Tony; Mathur, Harsh; Connor, Paula M. O’; Cotter, Paul D.; Ross, R. Paul; Hill, Colin

doi:10.1111/mmi.14183

Cited by 53 publications

(57 citation statements)

References 60 publications

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“…The nisin resistance protein (NSR) is a protease which cleaves nisin A at Ser29, significantly reducing the activity of the peptide. Employing a bioengineering strategy, Field et al (37) demonstrated that the substitution of residues 29 and 30 with proline and valine, respectively (derivative designated S29PV), rendered the peptide resistant to proteolytic digestion by NSR. In this study, we found that the nisin J producer displays a higher resistance to NSR proteolytic enzymes than does nisin A, which is possibly due to a glycine residue at position 30 instead of the isoleucine as found in nisin A. Interestingly, a study carried out by Simões et al (38) involving a multidrugresistant S. capitis clone, NRCS-A, a major pathogen involved in sepsis in preterm neonates, demonstrated the presence of an NSR-encoding gene.…”

Section: Discussionmentioning

confidence: 99%

Nisin J, a Novel Natural Nisin Variant, Is Produced by Staphylococcus capitis Sourced from the Human Skin Microbiota

et al. 2020

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The skin microbiota is thought to play a key role in host protection from infection. Nisin J is a novel nisin variant produced by Staphylococcus capitis APC 2923, a strain isolated from the toe web space area in a screening study performed on the human skin microbiota. Whole-genome sequencing and mass spectrometry of the purified peptide confirmed that S. capitis APC 2923 produces a 3,458-Da bacteriocin, designated nisin J, which exhibited antimicrobial activity against a range of Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) and Cutibacterium acnes. The gene order in the nisin J gene cluster (nsjFEGBTCJP) differs from that of other nisin variants in that it is lacking the nisin regulatory genes, nisRK, as well as the nisin immunity gene nisI. Nisin J has 9 amino acid changes compared to prototypical nisin A, with 8 amino acid substitutions, 6 of which are not present in other nisin variants (Ile4Lys, Met17Gln, Gly18Thr, Asn20Phe, Met21Ala, Ile30Gly, Val33His, and Lys34Thr), and an extra amino acid close to the C terminus, rendering nisin J the only nisin variant to contain 35 amino acids. This is the first report of a nisin variant produced by a Staphylococcus species and the first nisin producer isolated from human skin. IMPORTANCE This study describes the characterization of nisin J, the first example of a natural nisin variant, produced by a human skin isolate of staphylococcal origin. Nisin J displays inhibitory activity against a wide range of bacterial targets, including MRSA. This work demonstrates the potential of human commensals as a source for novel antimicrobials that could form part of the solution to antibiotic resistance across a broad range of bacterial pathogens.

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Section: Discussionmentioning

confidence: 99%

Nisin J, a Novel Natural Nisin Variant, Is Produced by Staphylococcus capitis Sourced from the Human Skin Microbiota

et al. 2020

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“…Induction experiments were performed using L. lactis NZ9000 pNZ8150 gfp+, where gfp acts as a reporter of expression from a nisin A inducible promoter 54 . An overnight culture of this strain was diluted 1:100 into fresh media and incubated until the OD 600 reached 0.5.…”

Section: Methodsmentioning

confidence: 99%

First evidence of production of the lantibiotic nisin P

García-Gutiérrez

O’Connor

Saalbach

et al. 2019

Preprint

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15Nisin P is a natural nisin variant, the genetic determinants for which were previously identified 16 in the genomes of two Streptococcus species, albeit with no confirmed evidence of production. 17 Here we describe Streptococcus agalactiae DPC7040, a human fecal isolate, which exhibits 18 antimicrobial activity against a panel of gut and food isolates by virtue of producing nisin P. 19 Nisin P was purified, and its predicted structure was confirmed by nanoLC-MS/MS, with both 20 the fully modified peptide and a variant without rings B and E being identified. Additionally, 21 we compared its spectrum of inhibition and minimum inhibitory concentration (MIC) with that 22 of nisin A and its antimicrobial effect in a fecal fermentation in comparison with nisin A and 23 H. We found that its antimicrobial activity was less potent than nisin A and H, and we propose 24 a link between this reduced activity and the peptide structure. 25 26 Keywords: nisin, antimicrobial, structure-activity relationship, fecal microbiome, 27 Structurally, nisin is classified as a lantibiotic because it contains lanthionine (Lan), an unusual 36 amino acid formed by two alanine residues linked by a sulphur atom through their β-carbon 4 . 37Other unusual amino acids present in nisin are dehydroalanine (Dha), dehydrobutyrine (Dhb) 38 3 3 and β-methyl-lanthionine 4 . Nisin activity and stability are closely related to its structure and 39 can be altered by pH changes. Increasing pH results in decreasing activity as a consequence of 40 alterations in structure, therefore, nisin is more stable at lower pH. Nisin is heat-stable and also 41 exhibits high stability at low temperatures, which makes it suitable for freeze-storage 7 . 42Nisin has nine reported natural variants ( Figure 1, Table 1). Nisin A is the most studied nisin 43 as it was the first one purified 8 . Nisin Z is considered the first natural variant of nisin A; it 44 differs in the presence of an asparagine amino acid in position 27 instead of a histidine residue 45 9 . This substitution has very little effect on antimicrobial activity, thermal and pH stability 46 compared to nisin A, but affects the solubility of the molecule, with nisin Z being more soluble 47 at neutral pH 10 . A study of its distribution also revealed that nisin Z is more widespread than 48 nisin A 10 . Nisin F, produced by L. lactis isolated from a fish gut, also has asparagine and valine 49 in positions 21 and 30 11 . Nisin Q was identified in an environmental L. lactis isolate and differs 50 from nisin A due to the presence of valine, leucine, asparagine and valine in positions 15, 21, 51 27 and 30, respectively 12 . Nisin U and U2 were the first variants found to be produced by a 52 species other than L. lactis 13 ; they are produced by Streptococcus uberis strain 42, isolated 53 from bovine mammary secretions 14 , and differ to nisin A by nine and ten amino acids, 54 respectively. Nisin H, produced by Streptococcus hyointestinalis was the first variant isolated 55 from a mammalian gastrointestin...

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“…Nisin (3.5 kDa) is produced by some Lactococcus lactis subsp. lactis strains and, as a protein compound, comprises 34 amino acid residues [8]. Its antimicrobial properties consist of permeabilization of the bacterial cell membrane by pore formation, leading to leakage of cellular content and disruption of the cell, preventing growth [9].…”

Section: Introductionmentioning

confidence: 99%

Nisin as a Novel Feed Additive: The Effects on Gut Microbial Modulation and Activity, Histological Parameters, and Growth Performance of Broiler Chickens

Kierończyk

Rawski

Mikołajczak

et al. 2020

Animals

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Two independent experiments were performed to evaluate the effect of nisin alone or with monensin on gut microbiota, gut microbial activities, and histomorphology (exp 1) and the effect of nisin application in a dose‒response manner on the growth performance of broiler chickens (exp 2). A total of 900 one-day-old female Ross 308 chicks (400, exp 1; 500, exp 2) were randomly distributed to four groups (exp 1; 10 replicate pens per treatment with 10 birds each), i.e., NA, no additives; MON, monensin (100 ppm); NIS, nisin (2700 IU/kg diet); and MON + NIS, a mixture of monensin (100 ppm) and nisin (2700 IU/kg diet); or 5 treatments (exp 2), NA, no additives; NIS100, nisin (100 IU/kg diet); NIS200, nisin (200 IU/kg diet); NIS400, nisin (400 IU/kg diet); and NIS800, nisin (800 IU/kg diet). Nisin supplementation positively affected the microbiota of the gut by reducing potentially pathogenic bacterial populations in the jejunum and ceca. The bacterial fermentation in the jejunum was significantly lowered by nisin addition. The addition of nisin from 100 IU to 800 IU decreased the FCR value over the entire experimental period. According to the results, nisin can be considered a natural dietary supplement for broiler chickens.

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Bioengineering nisin to overcome the nisin resistance protein

Cited by 53 publications

References 60 publications

Nisin J, a Novel Natural Nisin Variant, Is Produced by Staphylococcus capitis Sourced from the Human Skin Microbiota

Nisin J, a Novel Natural Nisin Variant, Is Produced by Staphylococcus capitis Sourced from the Human Skin Microbiota

First evidence of production of the lantibiotic nisin P

Nisin as a Novel Feed Additive: The Effects on Gut Microbial Modulation and Activity, Histological Parameters, and Growth Performance of Broiler Chickens

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