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Molecular methods for identification of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus using methionine biosynthesis and 16S rRNA genes.
J Dairy Res. 2008 Nov; 75(4):392-8.JD

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.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Cebeci A
Department of Biochemistry, Middle East Technical University, Turkey.
Gürakan GC
No affiliation info available

MeSH

DNA PrimersDNA, BacterialLactobacillus delbrueckiiMethioninePhenotypePolymerase Chain ReactionRNA, BacterialRNA, Ribosomal, 16SRestriction MappingStreptococcus thermophilusTurkeyYogurt

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18620618

Citation

Cebeci, Aysun, and G Candan Gürakan. "Molecular Methods for Identification of Lactobacillus Delbrueckii Subsp. Bulgaricus and Streptococcus Thermophilus Using Methionine Biosynthesis and 16S rRNA Genes." The Journal of Dairy Research, vol. 75, no. 4, 2008, pp. 392-8.
Cebeci A, Gürakan GC. Molecular methods for identification of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus using methionine biosynthesis and 16S rRNA genes. J Dairy Res. 2008;75(4):392-8.
Cebeci, A., & Gürakan, G. C. (2008). Molecular methods for identification of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus using methionine biosynthesis and 16S rRNA genes. The Journal of Dairy Research, 75(4), 392-8. https://doi.org/10.1017/S0022029908003543
Cebeci A, Gürakan GC. Molecular Methods for Identification of Lactobacillus Delbrueckii Subsp. Bulgaricus and Streptococcus Thermophilus Using Methionine Biosynthesis and 16S rRNA Genes. J Dairy Res. 2008;75(4):392-8. PubMed PMID: 18620618.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular methods for identification of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus using methionine biosynthesis and 16S rRNA genes. AU - Cebeci,Aysun, AU - Gürakan,G Candan, Y1 - 2008/07/14/ PY - 2008/7/16/pubmed PY - 2009/1/1/medline PY - 2008/7/16/entrez SP - 392 EP - 8 JF - The Journal of dairy research JO - J Dairy Res VL - 75 IS - 4 N2 - 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. SN - 0022-0299 UR - https://www.unboundmedicine.com/medline/citation/18620618/Molecular_methods_for_identification_of_Lactobacillus_delbrueckii_subsp__bulgaricus_and_Streptococcus_thermophilus_using_methionine_biosynthesis_and_16S_rRNA_genes_ DB - PRIME DP - Unbound Medicine ER -