Expression of soluble Saccharomyces cerevisiae alcohol dehydrogenase in Escherichia coli applicable to oxido-reduction bioconversions

被引:0
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作者
Pavol Utekal
Csaba Tóth
Anikó Illésová
Pavol Koiš
Lucia Bocánová
Ján Turňa
Hana Drahovská
Stanislav Stuchlík
机构
[1] Comenius University in Bratislava,Department of Molecular Biology, Faculty of Natural Sciences
[2] Comenius University in Bratislava,Department of Organic Chemistry, Faculty of Natural Sciences
来源
Biologia | 2014年 / 69卷
关键词
alcohol dehydrogenase; soluble protein expression; green notes bioconversion;
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摘要
Six-carbon aldehydes and alcohols belong to flavours and fragrances with wide application in the food, feed, cosmetic, chemical and pharmaceutical sectors. In the present study, we prepared the expression system for production of recombinant yeast alcohol dehydrogenase 1 (YADH1) from Saccharomyces cerevisiae which is suitable also for catalysis of the interconversion of C-6 aldehydes and alcohols. We have demonstrated that an effective three-step strategy can overcome the insolubility problems during YADH1 production in Escherichia coli. We used trxB and gor deficient expression strain, decreased concentration of isopropyl β-D-1-thiogalactopyranoside and lowered temperature to 20°C during induction. Finally, kinetic parameters of recombinant YADH1 were determined and we concluded it is a promising enzyme also for the interconversion of C-6 alcohols/aldehydes in green note volatile production.
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页码:722 / 726
页数:4
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