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Molecular methods for identification of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus using methionine biosynthesis and 16S rRNA genes

Published online by Cambridge University Press:  14 July 2008

Aysun Cebeci  and
G Candan Gürakan
Aysun Cebeci
Affiliation:
Department of Biochemistry, Middle East Technical University, Turkey
G Candan Gürakan*
Affiliation:
Department of Food Engineering, Middle East Technical University, Ankara, Turkey
*
*For correspondence; e-mail: [email protected]
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Abstract

Yoghurt and starter culture producers are still searching strains of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus to produce healthier yogurt with longer shelf life, better texture, taste and quality. However, selective identification of Lb. delbrueckii subsp. bulgaricus and Strep. thermophilus from a mixed population using microbiological and biochemical methods is difficult, time consuming and may not be accurate. In this study, a quick, sensitive and accurate method is proposed to identify both Lb. delbrueckii subsp. bulgaricus and Strep. thermophilus using PCR. The method is comprised of two parts. In the first part, methionine biosynthesis genes, known to be present in both species were partially amplified by designed primers (cysmet2F and cysmet2R). Partial amplification of the methionine biosynthesis gene which gives 700 bp fragment resulted in selective identification of Lb. bulgaricus and Strep. thermophilus. All 16 Lb. bulgaricus and 6 Strep. thermophilus isolates assessed by this method gave the expected amplification. On the other hand, further analysis of other closely related species with the same primers have indicated that the same product was also amplified in two more lactobacilli namely, Lb. delbrueckii subsp. lactis and Lb. helveticus species. Thus, in the second part of the method, further differentiation of Lb. delbrueckii subsp. bulgaricus and Strep. thermophilus from each other and these species was achieved using restriction analysis of 16S rRNA gene with EcoRI.

Keywords

Lb. delbrueckii subsp. bulgaricusStrep. thermophilusyoghurt starterstrain selection16S rRNA gene
Type
Research Article
Information
Journal of Dairy Research , Volume 75 , Issue 4 , November 2008 , pp. 392 - 398
Copyright
Copyright © Proprietors of Journal of Dairy Research 2008

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