Effects of Carbon Source on the Application of a Novel Foreign Gene Expression System in Saccharomyces cerevisiae Using the Upstream Region of the Candida Tropicalis Isocitrate Lyase Gene (UPR-ICL)
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概要
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The effects of various carbon sources were examined to optimize the application of a novel foreign gene expression system in Saccharomyces cerevisiae using the upstream region of the isocitrate lyase gene (UPR-ICL) of Candida tropicalis. A UPR-ICL-LacZ fusion gene was transformed into S. cerevisiae to compare the gene expression levels by measuring the β-galactosidase activity. Glucose, acetate, glycerol, lactate, and ethanol were used as carbon sources in different minimal media. All the carbon sources except for glucose led to a rapid increase in enzyme activity. Judging from the productivity and specific activity, acetate was the most effective carbon source for gene expression in the minimal media. At the same time, a high level of enzyme activity was also obtained in cells grown on glucose to later growth phases. This phenomenon was also observed in cells at the stationary phase in a rich glucose medium, and the productivity per unit volume of broth was the highest out of all the conditions examined. Determination of the glucose and ethanol concentrations in the media revealed that the increase in the level of gene expression promoted by UPR-ICL in S. cerevisiae was primarily dependent on glucose starvation, but not necessarily on ethanol accumulation, during cultivation. That is, gene expression is repressed until the stationary phase, where glucose is exhausted and the cell density becomes high. Immediately after glucose exhaustion, high levels of protein production were obtained. Accordingly, for rapid production of target proteins at high levels using this system on a laboratory scale, transferring cells cultivated in a glucose medium to a minimal acetate or ethanol medium seems effective.
- 社団法人日本生物工学会の論文
- 1995-12-25
著者
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Kurihara T
Laboratory Of Applied Biological Chemistry Department Of Synthetic Chemistry And Biological Chemistr
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Kurihara Tatsuo
Laboratory Of Microbial Biochemistry Institute For Chemical Research Kyoto University
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UEDA MITSUYOSHI
Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemist
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TANAKA ATSUO
Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemist
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Tanaka A
Department Of Etiology And Pathophysiology National Cardiovascular Center Research Institute
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Atomi H
Department Of Synthetic Chemistry And Biological Chemistry Graduate School Of Engineering Kyoto Univ
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Atomi Haruyuki
Laboratory Of Applied Biological Chemistry Department Of Synthetic Chemistry And Biological Chemistr
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UMEMURA KEN
Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemist
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KANAI TAMOTSU
Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemist
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TERANISHI YUTAKA
Yokohama Research Center, Mitsubishi Chemical Corporation
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Kurihara T
Kyoto Univ. Kyoto
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Umemura Ken
Laboratory Of Applied Biological Chemistry Department Of Synthetic Chemistry And Biological Chemistr
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Tanaka A
京都大学大学院工学研究科
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UEDA Mitsuyoshi
京都大学大学院工学研究科
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Atomi Haruyuki
Dept. Synth. Chem. Biol. Chem. Grad. Sch. Eng. Kyoto Univ.:crest Imanaka Project
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Tanaka A
Mie Univ. Mie Jpn
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Tanaka A
Kao Corp. Wakayama Jpn
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Takagi Atsuko
Department Of Etiology And Pathophysiology National Cardiovascular Center Research Institute
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Tanaka Atsuo
Laboratory Of Applied Biological Chemistry Department Of Synthetic Chemistry And Biological Chamistr
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Ueda Mitsuyoshi
Laboratory Of Applied Biological Chemistry Department Of Synthetic Chemistry And Biological Chemistr
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Ueda Mitsuyoshi
Laboratory Of Applied Biological Chemistry Department Of Synthetic Chemistry And Biological Chemistr
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Tanaka Atsuo
Laboratory Of Industrial Biochemistry Department Of Industrial Chemistry Faculty Of Engineering Kyot
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