2009The Journal of General and Applied MicrobiologyOpen access

Rapid identification of lactic acid bacteria isolated from home-made fermented milk in Tibet

Jie Yu, Zhihong Sun, Wenjun Liu, Jiachao Zhang, Tiansong Sun, Qiuhua Bao, Heping Zhang

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Abstract

Fermented milk is a very delicious and nutritional food distributed in the international markets. Rapid preliminary identification of lactic acid bacteria to the species level is an important issue for the fermentation industry. One hundred seventy-one strains of lactic acid bacteria (LAB) were isolated from twenty-nine kurut samples and fifteen traditional fermented mongolian cattle milk (FMCM) samples in Tibet, China. All of the strains were isolated and divided into bacilli or cocci by phenotype, and then differentiated by restriction fragment length polymorphism (RFLP) analysis using a set of restriction enzymes, AluI, HaeIII, BsmaI, TspRI and HinfI. Restriction pattern analyses indicated that the lactobacilli could be clearly identified at the species level and the LAB cocci at the genera level by the five restriction enzymes. Further studies on 16S rDNA sequences of representative and undefined strains showed that the combination of RFLP analysis of polymerase chain reaction (PCR) for amplifying 16S rDNA and 16S rDNA sequence analysis is rapid, easy to perform, and effective for large-scale preliminary identification of LAB.

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Fermented milk is a very delicious and nutritional food distributed in the international markets. Rapid preliminary identification of lactic acid bacteria to the species level is an important issue for the fermentation industry. One hundred seventy-one strains of lactic acid bacteria (LAB) were isolated from twenty-nine kurut samples and fifteen traditional fermented mongolian cattle milk (FMCM) samples in Tibet, China. All of the strains were isolated and divided into bacilli or cocci by phenotype, and then differentiated by restriction fragment length polymorphism (RFLP) analysis using a set of restriction enzymes, AluI, HaeIII, BsmaI, TspRI and HinfI. Restriction pattern analyses indicated that the lactobacilli could be clearly identified at the species level and the LAB cocci at the genera level by the five restriction enzymes. Further studies on 16S rDNA sequences of representative and undefined strains showed that the combination of RFLP analysis of polymerase chain reaction (PCR) for amplifying 16S rDNA and 16S rDNA sequence analysis is rapid, easy to perform, and effective for large-scale preliminary identification of LAB.

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Available abstract

Fermented milk is a very delicious and nutritional food distributed in the international markets. Rapid preliminary identification of lactic acid bacteria to the species level is an important issue for the fermentation industry. One hundred seventy-one strains of lactic acid bacteria (LAB) were isolated from twenty-nine kurut samples and fifteen traditional fermented mongolian cattle milk (FMCM) samples in Tibet, China. All of the strains were isolated and divided into bacilli or cocci by phenotype, and then differentiated by restriction fragment length polymorphism (RFLP) analysis using a set of restriction enzymes, AluI, HaeIII, BsmaI, TspRI and HinfI. Restriction pattern analyses indicated that the lactobacilli could be clearly identified at the species level and the LAB cocci at the genera level by the five restriction enzymes. Further studies on 16S rDNA sequences of representative and undefined strains showed that the combination of RFLP analysis of polymerase chain reaction (PCR) for amplifying 16S rDNA and 16S rDNA sequence analysis is rapid, easy to perform, and effective for large-scale preliminary identification of LAB.

Key concepts: HaeIII, Biology, 16S ribosomal RNA, Restriction fragment length polymorphism, Bacteria, Restriction enzyme, Bacilli, Fermentation

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