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Random amplified polymorphic DNA‐PCR based cloning of markers to identify the beer‐spoilage strains of Lactobacillus brevis , Pediococcus damnosus , Lactobacillus collinoides and Lactobacillus coryniformis
Author(s) -
Fujii T.,
Nakashima K.,
Hayashi N.
Publication year - 2005
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2005.02558.x
Subject(s) - lactobacillus brevis , biology , lactobacillus , rapd , food spoilage , microbiology and biotechnology , bacteria , pediococcus , lactic acid , genetics , lactobacillus plantarum , population , genetic diversity , demography , sociology
Aims: Beer‐spoilage ability of lactic acid bacteria such as Lactobacillus brevis is a strain‐dependent phenomenon in which the mechanism has not yet been completely clarified. In order to systematically identify genes that contribute to beer‐spoilage, large‐scale random amplified polymorphic DNA (RAPD)‐based cloning methods was carried out. Methods and Results: A systematic RAPD polymerase chain reaction (PCR) analysis using 600 primers was performed on beer‐spoilage and on nonspoilage strains of L. brevis . Among 600 primers, three were found to amplify a single locus highly specific to beer‐spoilage strains. DNA sequencing of this locus revealed a three‐part operon encoding a putative glycosyl transferase, membrane protein and teichoic acid glycosylation protein. PCR analysis of typical beer‐spoilage lactic acid bacteria suggested that this locus is highly specific to beer‐spoilage strains. Conclusion: The cloned markers are highly specific to identify the beer‐spoilage strains not only in L. brevis but also in Pediococcus damnosus , Lactobacillus collinoides and Lactobacillus coryniformis . Significance and Impact of the Study: This paper proves that RAPD‐PCR is an efficient method for cloning the strain‐specific genes from bacteria. The markers described here is one of the most useful tools to identify the beer‐spoilage strains of lactic acid bacteria.