Easy and Rapid Purification of Highly Active Nisin

Abts, André; Mavaro, Antonino; Stindt, Jan; Bakkes, Patrick J.; Metzger, Sabine; Driessen, Arnold J. M.; Smits, Sander H. J.; Schmitt, Lutz

doi:10.1155/2011/175145

Cited by 40 publications

(48 citation statements)

References 28 publications

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“…Nisin was basically purified as described in (Abts et al. ). In short, commercial available nisin powder (Sigma) was dissolved in 50 mmol/L lactic acid pH 3.…”

Section: Methodsmentioning

confidence: 99%

“…Nisin containing fractions were precipitated by TCA and dried after washing it with cold acetone (Abts et al. ). Upon usage nisin was dissolved in 50 mmol/L lactic acid (pH 3) and the concentration of nisin was measured by using RP‐HPLC (Abts et al.…”

Section: Methodsmentioning

confidence: 99%

“…The IC 50 value is the concentration of nisin were the growth of the L. lactis strain is inhibited by 50% as described in more detail earlier (Abts et al. ).…”

Section: Methodsmentioning

confidence: 99%

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The C‐terminus of nisin is important for the ABC transporter NisFEG to confer immunity in Lactococcus lactis

et al. 2014

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The lantibiotic nisin is a small 3.4 kDa antimicrobial peptide, which acts against Gram-positive bacteria in the nmol/L range. Nisin is produced and secreted by several Lactococcus lactis strains to ensure advantages against other bacteria in their habitat. Nisin contains five specific lanthionine rings of which the first two are important for Lipid II binding and the last two are crucial for the pore formation in the membrane. To gain immunity against nisin, the producing strain is expressing an ABC transporter called NisFEG, which expels nisin from the membrane. As a result six to eightfold more nisin is needed to affect the cells. The hydrolysis of ATP by NisFEG is required for this immunity as shown by a mutant, where the ATP hydrolysis is disrupted (NisFH181AEG). Furthermore, NisFEG recognizes the C-terminus of nisin, since deletion of the last six amino acids as well as of the last ring lowered the fold of immunity displayed by NisFEG.

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“…Nisin was basically purified as described in (Abts et al. ). In short, commercial available nisin powder (Sigma) was dissolved in 50 mmol/L lactic acid pH 3.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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The C‐terminus of nisin is important for the ABC transporter NisFEG to confer immunity in Lactococcus lactis

et al. 2014

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“…The active nisin was purified as described elsewhere (41). In brief, about 1.3 g of powder (corresponding to ϳ32 mg of nisin) was diluted in 100 ml of 50 mM lactic acid, pH 3, and filtered through a 0.45-m membrane filter (Pall Corporation).…”

Section: Methodsmentioning

confidence: 99%

Substrate Recognition and Specificity of the NisB Protein, the Lantibiotic Dehydratase Involved in Nisin Biosynthesis

Mavaro

Abts

Bakkes

et al. 2011

Journal of Biological Chemistry

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Nisin is a posttranslationally modified antimicrobial peptide containing the cyclic thioether amino acids lanthionine and methyllanthionine. Although much is known about its antimicrobial activity and mode of action, knowledge about the nisin modification process is still rather limited. The dehydratase NisB is believed to be the initial interaction partner in modification. NisB dehydrates specific serine and threonine residues in prenisin, whereas the cyclase NisC catalyzes the (methyl)lanthionine formation. The fully modified prenisin is exported and the leader peptide is cleaved off by the extracellular protease NisP. Light scattering analysis demonstrated that purified NisB is a dimer in solution. Using size exclusion chromatography and surface plasmon resonance, the interaction of NisB and prenisin, including several of its modified derivatives, was studied. Unmodified prenisin binds to NisB with an affinity of 1.05 ؎ 0.25 M, whereas the dehydrated and the fully modified derivatives bind with respective affinities of 0.31 ؎ 0.07 and 10.5 ؎ 1.7 M. The much lower affinity for the fully modified prenisin was related to a >20-fold higher off-rate. For all three peptides the stoichiometry of binding was 1:1. Active nisin, which is the equivalent of fully modified prenisin lacking the leader peptide did not bind to NisB, nor did prenisin in which the highly conserved FNLD box within the leader peptide was mutated to AAAA. Taken together our data indicate that the leader peptide is essential for initial recognition and binding of prenisin to NisB.

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“…Nisin works in the cytoplasmic membrane of the microorganism, by binding to the phospholipids, especially lipid II, an essential element in the cell wall (4,13,19,20).…”

Section: Introductionmentioning

confidence: 99%

Nisin-producing microorganisms and their implementation in brewers' wort

Müller-Auffermann

Grijalva²,

Jacob

et al. 2015

J. Inst. Brew.

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Nisin is a bacteriocin, which is capable of eliminating more than 90% of all potential beer spoilage Gram-positive bacteria. Hence, the implementation of nisin-producing cultures into the brewing process needs to be evaluated systematically. In this work, the genetic relationships and properties, as well as the reactions of four strains of Lactococcus lactis ssp. lactis, known to be capable of producing nisin (NCIMB 8780, 8586, 701402 and 701403), and seven isolated strains of the same species found in the beverage environment (TUM 575, 8947, 8127, 8446, 8673, 8973 and 8872), were tested under typical brewery conditions. As in previous work, it was found, that all of the tested strains could be genetically differentiated via PCR-(GTG) 5 . In addition, the absence of the genes HorA, HorC and ORF5 indicated that all strains were sensitive to hop components. The agar diffusion assay test proved to be the most reliable method to precisely determine different nisin concentrations. Nisin formation, acid formation and the reproduction rates of the organisms were tested subsequently in various brewery relevant culture media such as MRS, first wort (unhopped) and wort (hopped). MRS provided the best environment for bacterial growth and hence acid and nisin production. The four NCIMB strains, which were the only ones capable of producing nisin under the named conditions, were chosen for study with regards to their tolerance to specific compounds found in the brewery environment. The strains NCIMB 8780 and 8586 produced comparatively higher amounts of nisin and were more tolerant to hop bitterness, ethanol, high gravity and low extract conditions. The growth rates, acid production and nisin production of all strains decreased with increasing bitter units and ethanol content. The optimum extract concentration was 5-10°P.

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Easy and Rapid Purification of Highly Active Nisin

Cited by 40 publications

References 28 publications

The C‐terminus of nisin is important for the ABC transporter NisFEG to confer immunity in Lactococcus lactis

The C‐terminus of nisin is important for the ABC transporter NisFEG to confer immunity in Lactococcus lactis

Substrate Recognition and Specificity of the NisB Protein, the Lantibiotic Dehydratase Involved in Nisin Biosynthesis

Nisin-producing microorganisms and their implementation in brewers' wort

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