Lertwattanasakul Noppon, Sootsuwan Kaewta, Limtong Savitree, Thanonkeo Pornthap, Yamada Mamoru
Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, Japan.
Biosci Biotechnol Biochem. 2007 May;71(5):1170-82. doi: 10.1271/bbb.60622. Epub 2007 May 7.
Four genes encoding alcohol dehydrogenase (Adh) isozymes in the thermotolerant yeast Kluyveromyces marxianus, a potent candidate for ethanol production at high temperatures, were investigated. Of these, KmADH3 and KmADH4 were cloned and sequenced, and their deduced amino acid sequences were compared with those of KmAdh1 and KmAdh2 and other Adhs of Kluyveromyces lactis and Saccharomyces cerevisiae. The four KmAdhs had high sequence similarity, though KmAdh3 and KmAdh4 possessed an amino-terminal extension as a mitochondrial targeting sequence, and appear to belong to the zinc-containing Adh family. These results and the results of Southern blot experiments suggest that there are at least four Adh isozymes in K. marxianus, two cytoplasmic enzymes and two mitochondrial enzymes. The expression profile revealed that KmADH genes are differently expressed depending on growth phase and carbon source, suggesting that these highly homologous Adhs play distinctive roles in cells.
对高温酿酒酵母(Kluyveromyces marxianus)中编码乙醇脱氢酶(Adh)同工酶的四个基因进行了研究,该酵母是高温乙醇生产的有力候选菌株。其中,克隆并测序了KmADH3和KmADH4,并将其推导的氨基酸序列与KmAdh1和KmAdh2以及乳酸克鲁维酵母(Kluyveromyces lactis)和酿酒酵母(Saccharomyces cerevisiae)的其他Adh进行了比较。四个KmAdh具有高度的序列相似性,尽管KmAdh3和KmAdh4具有作为线粒体靶向序列的氨基末端延伸,并且似乎属于含锌Adh家族。这些结果以及Southern印迹实验的结果表明,马克斯克鲁维酵母中至少有四种Adh同工酶,两种细胞质酶和两种线粒体酶。表达谱显示,KmADH基因根据生长阶段和碳源的不同而表达不同,这表明这些高度同源的Adh在细胞中发挥着独特的作用。