The Location of Nisin in the Producer Organism, Streptococcus lactis

White, Richard; Hurst, A.

doi:10.1099/00221287-53-2-171

Cited by 29 publications

(17 citation statements)

References 14 publications

(17 reference statements)

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“…Glucosamine in S. Iactis is partly in the peptidoglycan and partly in a rhamnose polymer (22). It seems likely that the rhamnose polymer of the wall remained intact.…”

Section: Discussionmentioning

confidence: 99%

“…Fifty percent of the wall is composed of protein, and the wall also contains a rhamnose polymer (22); both these components are insensitive to lysozyme. When spheroplasted cells were incubated with proteolytic enzymes (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…The conversion of precursor to nisin occurs also on the surface (1 1). Further study of this process required protoplasts or spheroplasts of stationary phase cells but these cells are known to be lysozyme resistant (22).…”

Section: Introductionmentioning

confidence: 99%

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Preparation of spheroplasts from Streptococcus lactis

Kruse¹,

Hurst²

1972

Can. J. Microbiol.

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KRUSE, H., and A. H u~s r . 1972. Preparation of spheroplasts from Streptococcus lactis. Can. J. Microbiol. 18: 825-831. Stationary phase cells of Streptococcus lactis resisted lysis by lysozyrne under a variety of test conditions, although they lost 20y0 of their hexosarnine. Lysozyrne was bound by the cells and caused the optical density of the suspensions to rise. Incubation of the cells (0.5 rng dry weightlrnl) in buffer with lysozyrne (1 rng/rnl), NaCl (0.5 M ) , or sodium dodecylsulfate (0.1%) caused prompt lysis. The cells were stabilized with 10% polyethylene glycol but not by 10-30% sucrose. Lysozyrne and NaC1-treated cells in polyethylene glycol were spheroplasts because they were osmotically fragile, lost their shape but retained a rharnnose wall polymer. The spheroplasts retained all the DNA of the original cells, and 10yo of the original numbers could be cultured on agar. However, they lost all their rnurarnic acid and 60% of their hexosarnine. In contrast to NaCI, sodium dodecylsulfate-treated cells were killed. KRUSE, H., et A. H u~s r . 1972. Preparation of spheroplasts from Streptococcus lactis. Can. J. Microbiol. 18: 825-831. Les cellules de Streptococcus lactis en phase stationnaire rksistent a la lyse par la lysozyrne sous difftrentes conditions d'essai, bien qu'ils perdent 20% de leur hexosarnine. La lysozyrne ttant like par les cellules est responsable de l'augrnentation de la densitk optique des suspensions. On obtient une lyse rapide des cellules (0.5 rng poids sec/rnl) par incubation dans une solution tampon contenant de la lysozyrne (1 rnglrnl), NaCl (0.5 M ) ou du dodecylsulphate de sodium (0.1 %). On obtient une stabilisation des cellules avec lOy0 de polyethylene glycol rnais non avec 10-30yo de sucrose. Les cellules traittes a la lysozyrne et NaCl dans une solution de polyethylene glycol sont des sphkroplastes parce qu'elles sont sensibles la pression osrnotique; elles perdent leur forrne rnais retiennent un polyrnkre rharnnose de la paroi. Les sphkroplastes retiennent tout 1'ADN des cellules originales et 10yo de ces dernieres peuvent &re cultivkes sur gelose. Cependant, ils perdent la totalitk de leur acide rnurarnique et 60% d~ leur hexosarnine. Les cellules traitees au dodecylsulphate de sodium sont dttruites, contrairernent a celles traitkes au NaC1.

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“…Glucosamine in S. Iactis is partly in the peptidoglycan and partly in a rhamnose polymer (22). It seems likely that the rhamnose polymer of the wall remained intact.…”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Preparation of spheroplasts from Streptococcus lactis

Kruse¹,

Hurst²

1972

Can. J. Microbiol.

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“…Contemporary purification techniques for bacteriocins include chemical precipitation, separation through solvents used in combination with acid treatment of the culture followed by removal of the cells and then solvent extraction and precipitation, and high performance liquid chromatography or reverse phase chromatography. Currently, most methods rely on ammonium sulfate precipitation of the bacteriocins from cell‐free cultured broth.…”

Section: Introductionmentioning

confidence: 99%

Low molecular weight liquid media development for Lactobacilli producing bacteriocins

Zacharof

Lovitt

2012

J of Chemical Tech & Biotech

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BACKGROUND Contemporary purification techniques of Lactobacilli bacteriocins include chemical precipitation and separation through solvents to obtain highly potent semi‐purified bacteriocins. These methods are laborious and bacteriocin yields are low. To address this problem a set of new, efficient, cost effective media, was created, containing low molecular weight nutrient sources (LMWM). Using these media future separation and concentration of the desired metabolic products, using ultra‐ and nano‐filtration from the cultured broth was possible. RESULTS The LMWM were made through serial filtration (filters varying in pore size 30 kDa, 4 kDa and 1 kDa MWCO) of a modified optimum liquid medium for Lactobacilli growth. The developed media were tested for bacteriocin production and biomass growth, using three known bacteriocin‐producing Lactobacilli strains, Lactobacillus casei NCIMB 11970, Lactobacillus plantarum NCIMB 8014, Lactobacillus lactis NCIMB 8586. All were successfully grown (µmax 0.16 to 0.18 h−1) on the LMWM and produced a significant amount of bacteriocins in the range 110 to 130 IU mL−1. CONCLUSIONS LMWM do support Lactobacilli growth and bacteriocin production, establishing an alternative to the current production nutrient media. The uptake of the nutrient sources is facilitated as nitrogen sources, which were primarily responsible for growth, were supported in less complex forms. Copyright © 2012 Society of Chemical Industry

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“…A nisin precursor is first made ribosomally, followed by enzymatic modifications necessary to obtain the active antibiotic from the precursor molecule. The nisin-generating enzyme(s) and the antibiotic were located on the surface of the producer cells (6,12).…”

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confidence: 99%

Effect of Secondary Metabolites on the Organisms Producing Them: Effect of Nisin on Streptococcus lactis and Enterotoxin B on Staphylococcus aureus

Hurst¹,

Kruse²

1972

Antimicrob Agents Chemother

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The effect of secondary metabolites added to cultures of the organisms producing them was investigated. Nisin was added to growing cultures of a nisin-producing strain of Streptococcus lactis (354/07) Nisin is known to be synthesized after the log phase of growth (5), and recently we obtained evidence that it may be synthesized in at least two stages (6). A nisin precursor is first made ribosomally, followed by enzymatic modifications necessary to obtain the active antibiotic from the precursor molecule. The nisin-generating enzyme(s) and the antibiotic were located on the surface of the producer cells (6, 12).Surface synthesis of nisin might be necessary if the cells were sensitive to their own metabolic products. Also, if young cells were less able to exclude the toxic product from their internal environment than older cells, synthesis would have to be coordinated with the growth cycle. This could account for nisin synthesis occurring after the log phase of growth. We tested this hypothesis by adding nisin to cultures of the producer organism at different stages of the growth cycle and then followed growth turbidimetrically.Nisin was prepared by the method of Bailey and Hurst (1), and we also used a highly purified preparation donated by R. H. Hall of Aplin and Barrett Ltd., Yeovil, England. Nisin was estimated by a turbidimetric bioassay (4).We checked the purity of our preparations by a strongly dissociative disc-gel electrophoretic technique in 6 M urea (7). Our preparations were at least 95% pure; they gave a single major band with up to two additional faint bands detectable only at high concentration. The commercial and laboratory preparations had the same mobiljties.

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The Location of Nisin in the Producer Organism, Streptococcus lactis

Cited by 29 publications

References 14 publications

Preparation of spheroplasts from Streptococcus lactis

Preparation of spheroplasts from Streptococcus lactis

Low molecular weight liquid media development for Lactobacilli producing bacteriocins

Effect of Secondary Metabolites on the Organisms Producing Them: Effect of Nisin on Streptococcus lactis and Enterotoxin B on Staphylococcus aureus

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