Characterization of Achlya bisexualis β-Amylase Expression in an Amylolytic Industrial Strain of Saccharomyces cerevisiae
2008
Lee, O.H. (Chonnam National University, Gwangju, Republic of Korea) | Lim, M.H. (Chonnam National University, Gwangju, Republic of Korea) | Kim, J.H. (Chonnam National University, Gwangju, Republic of Korea) | Ryu, E.H. (Chonnam National University, Gwangju, Republic of Korea) | Ko, H.M. (Chonnam National University, Gwangju, Republic of Korea) | Chin, J.E. (Dongkang College, Gwangju, Republic of Korea) | Bai, S. (Chonnam National University, Gwangju, Republic of Korea), E-mail: [email protected]
To develop an amylolytic industrial yeast strain producing β-amylase, the BAMY gene encoding Achlya bisexualis β-amylase was constitutively expressed under the control of the alcohol dehydrogenase gene promoter (ADC1p) in an industrial strain of Saccharomyces cerevisiae. Yeast transformation was carried out by an integration system containing δ-sequences as the recombination site. The integrative cassette devoid of bacterial DNA sequences was constructed that contains the BAMY gene and δ-sequences. Industrial S. cerevisiae transformed with this integrative cassette secreted 45 kDa β-amylase into the culture medium. The β-amylase activity of the transformant was approximately 18.5-times higher than that of A. bisexualis. The multi-integrated BAMY genes in the transformant were stable after 100 generations of growth in nonselective medium. Hydrolysis of soluble starch and various starches with the enzyme released maltose but not glucose or oligosaccharides.
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