Physiological role of the glutathione-dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida boidinii
Physiological role of the glutathione-dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida boidinii
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DOI：
10.1099/00221287-148-9-2697
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发表时间：
2002-09-01
期刊：
影响因子：
2.8
通讯作者：
Kato, N
中科院分区：
文献类型：
--
作者：
Lee, B;Yurimoto, H;Kato, N
以甲醇为唯一碳源的甲醇营养酵母具有甲醛脱氢酶活性(FLD，EC 1.2.1.1)。该结构基因FLD1是从猪链霉菌基因组文库中克隆得到的。FLD1基因全长1263个碱基，内含子123个碱基，外显子编码380个氨基酸的基因产物，预测的氨基酸序列与其他生物的FLDS序列高度相似。FLD1基因在猪瘟病毒基因组中通过一步基因破坏而被破坏。在甲醇限制的恒化器培养条件下，fld1Delta菌株不能以甲醇为碳源生长，即使在低稀释率(D<；0.05h(-1))下也不能生长，而甲酸脱氢酶基因(FDH；另一种参与甲醛氧化途径的NADH产生脱氢酶)菌株可以存活。这些结果表明，FLD对C boidinii在甲醇上的生长是必需的，而不是FDH。
The methylotrophic yeast Candida boidinii exhibits formaldehyde dehydrogenase activity (FLD, EC 1.2.1.1) during growth on methanol as a sole carbon source. The structural gene, FLD1, was cloned from a genomic library of C. boidinii. The 1263 bp FLD1 gene contained a 123 bp intron and its exon encoded a gene product of 380 amino acids, whose predicted amino acid sequence showed high similarity to the sequences of FLDs from other organisms. The FLD1 gene was disrupted in the C boidinii genome by one-step gene disruption. The fld1Delta strain could not grow on methanol as a carbon source under methanol-limited chemostat culture conditions, even with low dilution rates (D < 0.05 h(-1)), whereas a strain with a disruption in the gene for formate dehydrogenase (FDH; another NADH-generating dehydrogenase involved in the formaldehyde oxidation pathway) could survive. These results indicated that FLD, but not FDH, is essential for growth of C boidinii on methanol.