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Cloning and Overexpression of the als, pflA, and adhB Genes in Streptococcus thermophilus and Their Effects on Metabolite Formation

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Abstract

Streptococcus thermophilus is a lactic acid bacterium and used as starter culture in the dairy industry, mainly in the manufacture of yoghurt, with Lactobacillus delbrueckii subsp. bulgaricus. It produces lactic acid as a major fermentation end product and some carbonyl compounds through sugar metabolism. The level of metabolites could be improved using molecular biotechnology. The genes of als, encoding α-acetolactate synthase (Als), the pflA, encoding pyruvate-formate lyase activating enzyme (PflA), and the adhB which encodes alcohol dehydrogenase (AdhB) of S. thermophilus NCFB2393 strain were amplified by polymerase chain reaction and separately cloned into the overexpression vector pNZ276 under the control of the lacA promoter. The strains were transformed individually with the constructed plasmids. Their abilities to generate important metabolites such as pyruvate, lactate, formate, acetaldehyde, acetoin, ethanol, and 2,3-butanediol in LM17 medium were analyzed using high-performance liquid chromatography. High level of 2,3-butanediol was obtained by overexpressing the als gene. The level of formate increased slightly by overexpressing the pflA gene. The overexpression of the adhB gene, on the other hand, resulted in a significant increase in the ethanol level.

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Acknowledgments

This work was supported by the Scientific and Technological Research Council of Turkey (TÜBİTAK), Project No: 110 O 218.

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Authors and Affiliations

  1. Biotechnology and Gene Engineering Laboratory, Agricultural Biotechnology Department, Agriculture Faculty, Kahramanmaras Sutcu Imam University, Avsar, 46060, Kahramanmaras, Turkey

    Ismail Akyol & Fatma Gul Ozcelik

  2. Food Technology Department, Yenisehir Vocational School, Uludag University, 16900, Yenisehir, Bursa, Turkey

    Asuman Karakas-Sen

  3. Animal Science Department, Agriculture Faculty, Kahramanmaras Sutcu Imam University, Avsar, 46060, Kahramanmaras, Turkey

    Emin Ozkose & M. Sait Ekinci

  4. Department of Food Engineering, Faculty of Agriculture, Kahramanmaras Sutcu Imam University, Avsar, 46060, Kahramanmaras, Turkey

    Yekta Gezginc

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  1. Ismail Akyol
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Correspondence to Ismail Akyol.

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Akyol, I., Ozcelik, F.G., Karakas-Sen, A. et al. Cloning and Overexpression of the als, pflA, and adhB Genes in Streptococcus thermophilus and Their Effects on Metabolite Formation. Mol Biotechnol 57, 923–930 (2015). https://doi.org/10.1007/s12033-015-9882-1

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  • DOI: https://doi.org/10.1007/s12033-015-9882-1

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