Sakai Y, Rogi T, Yonehara T, Kato N, Tani Y
Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University, Japan.
Biotechnology (N Y). 1994 Mar;12(3):291-3. doi: 10.1038/nbt0394-291.
Previous studies of ATP production with the methylotrophic yeast, Candida boidinii, suggested that the phosphorylation of AMP catalyzed by adenylate kinase (ADK) was rate-limiting. To investigate whether the enhancement of ADK activity in C. boidinii cells would improve ATP productivity, the Saccharomyces cerevisiae ADK1 gene encoding ADK was expressed in C. boidinii under the C. boidinii AOD1 promoter. Methanol-induced transformants had 10,000-fold enhanced levels of ADK activity and produced 23-fold more ATP from adenosine when compared to the control, parent strain. In a pH-controlled reaction system with successive adenosine-feeding, the ATP concentration in the reaction mixture reached 230 mM (117 g/l) over 45 hours, and was easily purified with an overall yield of 78 percent.
先前对甲基营养型酵母博伊丁假丝酵母（Candida boidinii）进行的ATP生成研究表明，由腺苷酸激酶（ADK）催化的AMP磷酸化是限速步骤。为了研究增强博伊丁假丝酵母细胞中ADK的活性是否会提高ATP的生产率，在博伊丁假丝酵母AOD1启动子的控制下，将编码ADK的酿酒酵母ADK1基因在博伊丁假丝酵母中进行表达。与对照亲本菌株相比，甲醇诱导的转化体的ADK活性提高了10000倍，并且从腺苷产生的ATP增加了23倍。在一个连续供应腺苷的pH受控反应体系中，反应混合物中的ATP浓度在45小时内达到230 mM（117 g/L），并且易于纯化，总产率为78%。
