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Analysis of the α-amylase gene sequence and the enzyme activity of Indian rock oyster Saccostrea forskali

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  • Chemistry and Biochemistry
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Abstract

Indian rock oyster Saccostrea forskali is an important commercial species in Thailand. In this study, its full-length α-amylase (SfAmy) cDNA nucleotide sequence was investigated. The SfAmy cDNA was 1,689 bp long and contained a 1,563-bp open reading frame encoding 520 amino acid residues, including a 17-amino acid signal peptide. The molecular mass and the estimated isoelectric point (pI) of the deduced mature S. forskali α-amylase (SfAMY) were 55.948 kDa and 6.45, respectively. The deduced protein sequence showed 45–88 % identity to other mollusk AMYs. The molecular weight was confirmed by the weight of the purified native enzyme. The specific activities of crude and purified native enzymes toward 1 % starch were 29.53 and 187.42 U/mg. In addition, the obtained recombinant SfAMY also showed activity in digesting 1 % starch. The specific activities of the crude and purified recombinant proteins were 11.8 and 46 U/mg. Both enzymes showed optimal activity temperature at 40 °C but their optimum pH values were different, 6.0 for the native and 5.0 for the recombinant. The expression of SfAmy examined by RT-PCR showed the highest levels in the digestive gland but none was observed in the adductor muscle.

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Acknowledgments

We are grateful to Dr. Nora Fascestti for critical reading of manuscript. We thank the Higher Education Commission of Thailand and the Center of Agricultural Biotechnology, Kasetsart University, Kampaengsaen Campus, Nakon Pathom, for the grant to TT.

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Authors and Affiliations

  1. Center for Agricultural Biotechnology, Kasetsart University, Kampaengsaen Campus, Nakhon Pathom, Thailand

    Thanaset Thongsaiklaing

  2. Center for Excellence on Agricultural Biotechnology (AG-BIO/PERDO-CHE), Bangkok, Thailand

    Thanaset Thongsaiklaing

  3. Interdisciplinary Graduate Program in Genetic Engineering, Kasetsart University, Bangkok, Thailand

    Wimonsiri Sehawong

  4. Department of Genetics, Faculty of Science, Kasetsart University, Bangkok, Thailand

    Anchanee Kubera & Lertluk Ngernsiri

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  1. Thanaset Thongsaiklaing
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  2. Wimonsiri Sehawong
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  3. Anchanee Kubera
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  4. Lertluk Ngernsiri
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Correspondence to Lertluk Ngernsiri.

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T. Thongsaiklaing and W. Sehawong contributed equally to this work.

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Thongsaiklaing, T., Sehawong, W., Kubera, A. et al. Analysis of the α-amylase gene sequence and the enzyme activity of Indian rock oyster Saccostrea forskali . Fish Sci 80, 589–601 (2014). https://doi.org/10.1007/s12562-014-0708-z

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  • DOI: https://doi.org/10.1007/s12562-014-0708-z

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