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Biotechnology Techniques to Characterize and Enhance the Antimicrobial Properties of a Novel Food Biopreservative


To study the biochemistry and genetics of a novel bacteriocin, a natural antibacterial peptide, produced by Streptococcus sp. ATCC 10035. To characterize the structure and genetics of the bacteriocin two primary phases of research were to be conducted: <ol> <li>The determination of the primary structure of the bacteriocin,
<li>The isolation and characterization of the gene producing the bacteriocin. </ol>
This study compliments a program in which a bacteriocin (diplococcin) produced by Streptococcus sp. ATCC 10035 is be used as a biopreservative in a novel hurdle system for shelf stable ration components. The goal of the program is to clone the bacteriocin gene and express an active peptide with enhanced properties.

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The Streptococcus sp. ATCC 10035 bacteriocin has been isolated and partially characterized. Approximately 16 peptides, ranging in size from 5.5kb to 67kb, were isolated from the bacterial extract. Reverse phase HPLC and an electroblot transfer technique to a membrane were used to purify the peptide for amino acid composition and sequence analysis. The amino acid composition data was very different from what we expected from the results.
It has been reported that Streptococcus sp. ATCC 10035 is a diplococcin producer however we found the compositional data to be very different from the reported compositional data of diplococcin. The presence of lanthionine, a modified amino acid, indicated we had an entirely different class of bacteriocin then the expected diplococcin class of bacteriocin. In addition, the N-terminal sequence data indicated that we had isolated a unique bacteriocin that had not yet been identified or characterized. The genetics work is currently underway with only preliminary data available.

Muller, Wayne
US Army - Soldier and Biological Chemical Command
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