Passoth V, Schäfer B, Liebel B, Weierstall T, Klinner U
Institut für Biologie IV (Mikrobiologie), RWTH Aachen, Germany.
Yeast. 1998 Oct;14(14):1311-25. doi: 10.1002/(SICI)1097-0061(1998100)14:14<1311::AID-YEA315>3.0.CO;2-T.
Two Pichia stipitis ADH genes (PsADH1 and PsADH2) were isolated by complementation of a Saccharomyces cerevisiae Adh(-)-mutant. The genes enabled the transformants to grow in the presence of antimycin A on glucose, to use ethanol as sole carbon source and made them sensitive to allylalcohol. The sequences of the genes showed similarities of 70-77% to sequences of ADH genes of Candida albicans, Kluyveromyces lactis, K. marxianus, and S. cerevisiae and about 60% homology to those of Schizosaccharomyces pombe and Aspergillus flavus. Southern hybridization experiments suggested that P. stipitis has only these two ADH genes. Both genes are located on the largest chromosome of P. stipitis. PsADH2 encodes for the ADH activity that is responsible for ethanol formation at oxygen limitation. The gene is regulated at the transcriptional level. Moreover, also in cells grown on ethanol, only PsADH2 transcript was found. PsADH1 transcript was detected under aerobic conditions on fermentable carbon sources.
通过互补酿酒酵母Adh(-)突变体分离出了两个树干毕赤酵母ADH基因（PsADH1和PsADH2）。这些基因使转化体能够在抗霉素A存在的情况下在葡萄糖上生长，以乙醇作为唯一碳源，并使它们对烯丙醇敏感。这些基因的序列与白色念珠菌、乳酸克鲁维酵母、马克斯克鲁维酵母和酿酒酵母的ADH基因序列具有70-77%的相似性，与粟酒裂殖酵母和黄曲霉的ADH基因序列具有约60%的同源性。Southern杂交实验表明树干毕赤酵母仅具有这两个ADH基因。这两个基因都位于树干毕赤酵母的最大染色体上。PsADH2编码在氧限制条件下负责乙醇形成的ADH活性。该基因在转录水平上受到调控。此外，在以乙醇为碳源生长的细胞中，也只发现了PsADH2转录本。在需氧条件下，在可发酵碳源上检测到了PsADH1转录本。
