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Novel Bacteriocins, Transport and Vector System and Method of Use Thereof

a technology of bacteriocin and transport vector, applied in the field of new bacteriocins, bacteriocins, immunity genes, can solve the problems of severe spoilage problems, meat lactics, extended storage life of vacuum-packaged meat, etc., to inhibit bacteria infection of fermentation reactors and inhibit spoilage bacteria

Inactive Publication Date: 2010-05-27
STILES MICHAEL E +10
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The invention is about new bacteriocins (antimicrobial peptides) called brochocin-C and enterocin-900, and their corresponding immunity peptides. The invention also includes a method for inhibiting pathogenic bacteria by using these bacteriocins. Additionally, the invention includes an expression vector for obtaining secretion of proteins from lactics, and a novel food-grade plasmid that can be used as a plasmid vector for genes including bacteriocins. The invention also includes a method to preserve food by adding bacteriocin-producing bacteria.

Problems solved by technology

Vacuum packaged meats have an extended but unpredictable storage life dependent on the types of lactics that dominate the microflora.
Meat lactics can cause severe spoilage problems, such as sulphide odors or greening by some Lactobacillus species and gas or slime production by Leuconostoc species.
Nisin is unusual because it is active 5 against a wide range of gram-positive bacteria, including the spores of Clostridium botulinum; unfortunately, its producer strain does not grow in chill-stored meats, and nisin does not function in meat systems.

Method used

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  • Novel Bacteriocins, Transport and Vector System and Method of Use Thereof
  • Novel Bacteriocins, Transport and Vector System and Method of Use Thereof
  • Novel Bacteriocins, Transport and Vector System and Method of Use Thereof

Examples

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example 1

Bacteriocins, Sources, Methods to Propagate

[0102]Table 1 describes many different bacterial strains and plasmids, the bacteriocins they contain and references which provide additional information about the bacterocin or bacterial strain. For information on the best method to grow a particular organism refer to the appropriate reference or reference therein.

example 2

Method to Use Signal Peptide to Direct the Secretion of Substrates Example Using Divergicin A Signal Peptide and Carnobacteriocin B2 as Substrate

[0103]Bacterial strains and media. Bacterial strains and plasmids used in this study are listed in Table 1. Carnobacteria were grown in APT broth (Difco Laboratories, Detroit, Mich.) at 25° C. without agitation. E. coli was grown in Luria Bertani (LB) medium at 37° C. on a rotary shaker. Agar plates were made by addition of 1.5% (wt / vol) agar to broth media. Antibiotics were added as selective agents when appropriate, as follows: erythromycin 200 μg ml-1 and ampicillin 100 μg ml-1 for E. coli and erythromycin 10 μg ml-1 for carnobacteria. Stock cultures of the bacterial strains were stored at −70° C. in the appropriate broth containing 20% (vol / vol) glycerol.

[0104]Oligonucleotide primer synthesis and amplification reactions. In the 3′ region of the nucleotide sequence encoding the signal peptide of divergicin A there is a HindIII restrictio...

example 3

Method to Use Leader Peptides to Direct Secretion of Substrates Via Dedicated Transport System

[0114]Bacterial strains and media. C. divergens LV13 (Worobo at al., 1995), C. divergens UAL278 (McCormick et al., unpublished), L. gelidum 187-13 and L. gelidum 187-22 (Hastings and Stiles, 1991), and Pediococcus pentosaceus FBB63C (Graham and McKay, 1985) were grown in APT broth (All Purpose Tween; Difco Laboratories Inc.) at 25° C. and 30° C., respectively. L. lactis IL1403 (Chopin at al., 1984) and L. lactis IL1403(pMB500) (van Belkum et al., 1989) were grown in glucose-M17 broth (Terzaghi and Sandine, 1975) at 30 E. coli strains MH1 (Casadaban and Cohen, 1980), DH5((BRL Life Technologies Inc.), BL21(DE3) (Studier and Moffat, 1986), MC4100 (Casadaban, 1976), and ZK796 (Wandersman and Delepelaire, 1990) were grown in TY broth at 37° C. (Rottlander and Trautner, 1970), Solid plating media were prepared by adding 1.2% (wt / vol) agar to the broth media. C. divergens UAL278 cells propagated o...

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PUM

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Abstract

New bacteriocins capable of inhibiting the growth of bacteria are disclosed, along with methods of obtaining secretion of proteins from lactic acid bacteria, and methods for protecting foodstuffs.

Description

[0001]This application is a divisional of 09 / 883,343 filed Jun. 19, 2001; which is a continuation of Ser. No. 08 / 924,629 filed Sep. 5, 1997 (now is U.S. Pat. No. 6,403,082); which claims the benefit of 60 / 026,257 filed Sep. 5, 1996.FIELD OF INVENTION[0002]This invention relates to novel polypeptides, bacteriocins, immunity genes obtained from lactic acid bacteria and a method of use thereof.BACKGROUND: OF THE INVENTION[0003]With the current consumer demand for fresh (i.e., never frozen) foods, it is important that methods be developed for safe storage of these products especially for fresh meats which are manufactured locally but are marketed around the world. The lactic microflora (lactic acid bacteria) of vacuum packaged meats delays spoilage for weeks or months, as opposed to meats packaged under aerobic conditions which develop a putrefactive microflora that causes spoilage within days.[0004]Vacuum packaged meats have an extended but unpredictable storage life dependent on the t...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12P21/00C12N15/63C12N1/00A23L1/30A01N37/18A23B4/22A23L1/305A23L3/3571A61K38/00A61P31/00A61P31/04C07K14/31C07K14/315C07K14/335C07K14/34C07K16/12C12N1/21C12N15/09C12P21/02C12P21/08G01N33/569
CPCA23B4/22A23L3/3571A61K38/00C07K14/31G01N2333/315C07K14/335C07K14/34C07K2319/02G01N33/56916C07K14/315A61P31/00A61P31/04
Inventor STILES, MICHAEL E.VEDERAS, JOHN CVAN BELKUM, MARIUS JWOROBO, RANDY WWOROBO, RODNEY JMCCORMICK, JOHN KGREER, GORDON G.MCMULLEN, LYNN MLEISNER, JORGEN JPOON, ALISONFRANZ, CHARLES M.A.P.
Owner STILES MICHAEL E
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