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Cloning and expression of the gene encoding Streptomyces coelicolor A3(2) α-galactosidase belonging to family 36

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Abstract

The α-galactosidase gene of Streptomyces coelicolor A3(2) was cloned, expressed in Escherichia coli and characterized. It consisted of 1497 nucleotides encoding a protein of 499 amino acids with a predicted molecular weight of 57,385. The observed homology between the deduced amino acid sequences of the enzyme and α-galactosidase from Thermus thermophilus was over 40%. The α-galactosidase gene was assigned to family 36 of the glycosyl hydrolases. The enzyme purified from recombinant E. coli showed optimal activity at 40 °C and pH 7. The enzyme hydrolyzed p-nitrophenyl-α-D-galactopyroside, raffinose, stachyose but not melibiose and galactomanno-oligosaccharides, indicating that this enzyme recognizes not only the galactose moiety but also other substrates.

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Authors and Affiliations

  1. Graduated school of Life and Environmental Science, University of Tsukuba, Ibaraki, 305-0006, Japan

    Kenji Kondoh & Kohei Morisaki

  2. Biological Function Division, National Food Research Institute, Tsukuba, Ibaraki, 305-8642, Japan

    Wook-Dong Kim, Satoshi Kaneko & Hideyuki Kobayashi

  3. Departmant of Food Engineering and Biotechnology, Kyouguan University, Kyunggi-do, 461-701, Korea

    Gwi-Gun Park

Authors
  1. Kenji Kondoh
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  2. Kohei Morisaki
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  3. Wook-Dong Kim
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  4. Gwi-Gun Park
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  5. Satoshi Kaneko
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  6. Hideyuki Kobayashi
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Correspondence to Hideyuki Kobayashi.

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Kondoh, K., Morisaki, K., Kim, WD. et al. Cloning and expression of the gene encoding Streptomyces coelicolor A3(2) α-galactosidase belonging to family 36. Biotechnol Lett 27, 641–647 (2005). https://doi.org/10.1007/s10529-005-3660-2

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  • DOI: https://doi.org/10.1007/s10529-005-3660-2

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