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Assessment of expression of Leloir pathway genes in wild-type galactose-fermenting Streptococcus thermophilus by real-time PCR

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Abstract

In this study, four galactose-positive (Gal+) Streptococcus thermophilus strains viz. AJM, JM1, KM3 and AUKD8 and one galactose-negative (Gal−) S. thermophilus NCDC 218 were used to characterize the organization of Leloir pathway genes using long-range PCR, and expression of these genes were studied using real-time PCR, in presence of different sugars. Long-range PCR results showed that both Gal+ and Gal− isolates, the gal–lac gene order (galRKTEM–lacSZ), are conserved including the size of individual genes. The promoter sequence of the three Gal+ isolates (AJM, JM1 and KM3) possessed single base pair deletion at −28 region of galR and C to T substitution at −9 box galK region. In contrast, Gal+ AUKD8 had A to T substitution at preceding −25 region of galR. The expression of galK and galM grown in the presence of galactose was significantly higher in case of AJM (30- and 7.6-fold, respectively), followed by KM3 and JM1. In addition, galR, galT and galE showed higher expression in galactose, than in lactose and glucose medium. This study gives a preliminary idea on Leloir pathway gene expression in wild Gal+ S. thermophilus, and further studies may throw more light on the role of gal–lac operon in galactose metabolism.

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Acknowledgments

We acknowledge Dr. Sachinandan De, Biswajit Brahma and Ayan Mukherjee (Animal Biotechnology Division, National Dairy Research Institute) for their suggestions and providing laboratory facilities at different stages of this work. The research of first author has been made possible by fellowship of the Indian Council of Agricultural Research (ICAR, India).

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This article does not contain any studies with human or animal subjects.

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Authors and Affiliations

  1. Dairy Microbiology Division, National Dairy Research Institute, Karnal, 132001, India

    Kaliyaperumal Anbukkarasi, Dhiraj Kumar Nanda, Thiyagamoorthy UmaMaheswari, Prashant Singh & Rameshwar Singh

  2. Department of Bioproducts, Central Leather Research Institute, Chennai, 600020, India

    Thiagarajan Hemalatha

  3. Department of Food Science, University of Missouri, Columbia, MO, 65211, USA

    Prashant Singh

  4. Directorate of Knowledge Management in Agriculture, Indian Council of Agricultural Research, New Delhi, 110012, India

    Rameshwar Singh

Authors
  1. Kaliyaperumal Anbukkarasi
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  2. Dhiraj Kumar Nanda
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  3. Thiyagamoorthy UmaMaheswari
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  4. Thiagarajan Hemalatha
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  5. Prashant Singh
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  6. Rameshwar Singh
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Correspondence to Rameshwar Singh.

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Anbukkarasi, K., Nanda, D.K., UmaMaheswari, T. et al. Assessment of expression of Leloir pathway genes in wild-type galactose-fermenting Streptococcus thermophilus by real-time PCR. Eur Food Res Technol 239, 895–903 (2014). https://doi.org/10.1007/s00217-014-2286-9

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  • DOI: https://doi.org/10.1007/s00217-014-2286-9

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