Construction of an amylolytic industrial strain of Saccharomyces cerevisiae containing the Schwanniomyces occidentalis alpha-amylase gene.
The gene encoding Schwanniomyces occidentalis alpha-amylase (AMY) was introduced into the chromosomal delta sequences of an industrial strain of Saccharomyces cerevisiae. To obtain a strain suitable for commercial use, an delta-integrative cassette devoid of bacterial DNA sequences was constructed that contains the AMY gene and aureobasidin A resistance gene (AUR1-C) as the selection marker. The AMY gene was expressed under the control of the alcohol dehydrogenase gene promoter (ADC1p). The alpha-amylase activity of Sacc. cerevisiae transformed with this integrative cassette was 6 times higher than that of Sch. occidentalis. The transformants (integrants) were mitotically stable after 100 generations in nonselective medium.
Kang
Na-Young
NY
Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University, Gwangju 500-757, Korea.
Park
Jeong-Nam
JN
Chin
Jong-Eon
JE
Lee
Hwanghee Blaise
HB
Im
Suhn-Young
SY
Bai
Suk
S
D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D007218:Industrial Microbiology / Q000379:methods*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D004718:Saccharomycetales / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D000516:alpha-Amylases / Q000096:biosynthesis* / Q000235:genetics*
D011994:Recombinant Proteins; D000516:alpha-Amylases