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KMID : 0545119920020020115
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Journal of Microbiology and Biotechnology
1992 Volume.2 No. 2 p.115 ~ p.121
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Molecular Cloning and Expression of α-Amylase Gene from Bacillus stearothermophilus in Zymomonas mobilis ZM4
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SONG KI-BANG
RHEE SANG-KI
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Abstract
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In order to broaden the spectrum of substrate utilization of a Gram negative bacterium Zymomonas mobilis which has a great potential as an industrial ethanol producing microorganism, cloning of α-amylase gene into Z. mobilis ZM4 was tried. The α-amylase gene was isolated from Bacillus stearothermophilus. By Southern blot analysis, it was proven that the α-amylase gene fragment was originated from a naturally occuring plasmid of B. stearothermophilus ATCC31195. To place α-amylase gene under the control of Z. mobilis promoter, two different Z. mobilis expression vectors, pZA26 and pLOI204, were used. The truncated α-amylase gene was then introduced into these vectors. Both qualitative and quantitative activities of α-amylase were observed in Z. mobilis cells harboring these plasmids with the α-amylase gene inserted. Gas chromatographic analysis of ethanol showed that one of the Z. mobilis transconjugants was capable of producing 67 mM ethanol from rich medium(RM) containing 5% soluble starch as a sole carbon source.
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