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Probiotic strains from Lactobacillus salivarius and antimicrobial agents obtained therefrom

Inactive Publication Date: 2004-02-26
ENTERPRISE IRELAND & UNIV COLLEGE CORK +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0050] The above mentioned properties of ABP118, namely pH, temperature and storage stability mean that this bacteriocin is likely to be of value in the food and pharmaceutical industries.

Problems solved by technology

Controversy exists over many of these beneficial effects as no standardised procedures are available and contradictory results have been published with regard to the possible beneficial effects of cultured products containing `probiotic` bacteria.
However, these strains are not indigenous to the infected host species, which is one of the criteria which is required for a successful probiotic strain for human use.

Method used

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  • Probiotic strains from Lactobacillus salivarius and antimicrobial agents obtained therefrom
  • Probiotic strains from Lactobacillus salivarius and antimicrobial agents obtained therefrom
  • Probiotic strains from Lactobacillus salivarius and antimicrobial agents obtained therefrom

Examples

Experimental program
Comparison scheme
Effect test

example 2

[0095] Source and Maintenance of Strains

[0096] The Lactobacillus strains used were isolated as described in Example 1.

[0097] Human Gastric Juice

[0098] Human gastric juice was obtained from healthy subjects by aspiration through a nasogastric tube (Mercy Hospital, Cork, Ireland). It was immediately centrifuged at 13,000 g for 30 min to remove all solid particles, sterilised through 0.45 .mu.m and 0.2 .mu.m filters and divided into 40 ml aliquots which were stored at 4.degree. C. and -20.degree. C.

[0099] The pH and pepsin activity of the samples were measured prior to experimental use. Pepsin activity was measured using the quantitative haemoglobin assay (Gautam, S. and de La Motte, R. S., (1989) Proteolytic enzymes, a practical approach. Chapter 3. R. J. Beynon and J. S. Bond (eds.), IRL Press, Oxford University Press; (Dawson, R. M. (1969) In Data for Biochemical Research 138. R. M. Dawson, D. C. Elliot and K. M. Jones (eds.), Clarendon Press, Oxford). Briefly, aliquots of gastric j...

example 3

[0118] Lactobacillus strains used were isolated as described in Example 1.

[0119] Growth of Cultures in the Presence of Bovine and Porcine Bile

[0120] Fresh cultures were streaked onto MRS / TPY agar plates supplemented with bovine bile (B-8381, Sigma Chemical Co. Ltd., Poole) at concentrations of 0.3, 1.0, 1.5, 5.0 and 7.5% (w / v) and porcine bile (B-8631, Sigma Chemical Co. Ltd., Poole) at concentrations of 0.3, 0.5, 1.0, 1.5, 5.0 and 7.5% (w / v). Plates were incubated at 37.degree. C. under anaerobic conditions and growth was recorded after 24-48 h.

[0121] Growth of Cultures in the Presence of Human Bile

[0122] Bile samples, isolated from several human gall-bladders, were stored at -80.degree. C. before use. For experimental work, bile samples were thawed, pooled and sterilised at 80.degree. C. for 10 min. Bile acid composition of human bile was determined using reverse-phase HPLC in combination with a pulsed amperometric detector according to the method of (Dekker, R. R. et al., (1991) ...

example 4

[0148] Growth and Maintenance of Cultures

[0149] The microorganisms screened for antimicrobial production were the seventeen strains listed in Table 3 and were isolated from the human intestinal tract as described in Example 1. All strains belonged to the UCC culture collection

[0150] Cultures were maintained as frozen stocks at -20.degree. C. in the appropriate growth medium and 40% glycerol. Lactobacilli were routinely cultured in deMann Rogosa Sharpe (MRS) medium at 37.degree. C. under strict anaerobic conditions (BBL Gas Jars using the Merck Anaerocult A Gas Pak system).

[0151] The indicator microorganisms used in this Example, many of which are wildtype strains isolated in the Mercy Hospital, Cork, Ireland, were propagated in the following medium under the following growth conditions: Staphylococcus (37.degree. C., aerobic), Bacillus (37.degree. C., aerobic), Pseudomonas (30.degree. C., aerobic), Escherichia coli (37.degree. C., anaerobic), Salmonella (37.degree. C., anaerobic) an...

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Abstract

A strain of Lactobacillus salivarius isolated from resected and washed human gastrointestinal tract inhibits a broad range of Gram positive and Gram negative microorganisms and secretes a product having antimicrobial activity into a cell-free supernatant. The activity is produced only by growing cells and is destroyed by proteinase K and pronase E, the inhibitory properties of the strain and its secretory products being maintained in the presence of physiological concentrations of human bile and human gastric juice. The strain exhibits a broad-spectrum of activity against bacteria including Listeria, Staphylococcus, including methocillin resistant St. aureus (MRSA), and Bacillus, but does not inhibit many closely related lactobacilli. An antimicrobial agent is obtained from the strain which has bacteriocin-like properties.

Description

[0001] This application is a divisional of co-pending application Ser. No. 09 / 367,105, filed on Nov. 10, 1999, and for which priority is claimed under 35 U.S.C. .sctn. 120. application Ser. No. 09 / 367,105 is the national phase o f PCT International Application No. PCT / IE98 / 00010 filed on Feb. 11, 1998, under 35 U.S.C. .sctn. 371. The entire contents of each of the above-identified applications are hereby incorporated by reference. This application also claims priority of International Application No. PCT / IE97 / 00007 filed on Feb. 11, 1997, under 35 U.S.C. .sctn. 119.[0002] This invention relates to probiotic bacterial strains capable of producing antimicrobial agents which have various applications in food stuffs and in medicine. More particularly, the invention relates to probiotic strains of Lactobacillus salivarius and to a peptide antibacterial agent derived therefrom with bacteriocin-like properties.[0003] Much research has been carried out in the field of human probiotics in th...

Claims

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Application Information

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IPC IPC(8): A23L1/28A23C9/12A23L29/00A61K35/74A61K35/747A61K38/00A61P31/04C07K14/335C12N1/00C12N1/20C12N15/09C12R1/225
CPCA23L1/0345A23Y2220/77C12R1/225C07K14/335A61K35/747A23L29/065C12N1/00A61P31/04C12R2001/225C12N1/205A23V2400/179
Inventor COLLINS, JOHN KEVINO'SULLIVAN, GERALD CHRISTOPHERTHORNTON, GERARDINE MARYO'SULLIVAN, MARIAN MARY GERALDINE
Owner ENTERPRISE IRELAND & UNIV COLLEGE CORK
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