Hirasawa Takashi, Ookubo Aki, Yoshikawa Katsunori, Nagahisa Keisuke, Furusawa Chikara, Sawai Hideki, Shimizu Hiroshi
Department of Bioinformatic Engineering, Osaka University, Suita, Japan.
Appl Microbiol Biotechnol. 2009 Oct;84(6):1149-59. doi: 10.1007/s00253-009-2209-z.
In order to determine whether transcriptome data obtained by DNA microarray analysis could be used to identify the genes involved in target metabolite production, we tried to identify the genes involved in L-lactate production by L-lactate-producing recombinant Saccharomyces cerevisiae strains. We obtained DNA microarray data for these strains. Plasmids carrying lactic acid bacteria, bovine, and human L-lactate dehydrogenase (LDH) genes were introduced into PDC1-disrupted S. cerevisiae strains. L-Lactate productivity of the strains harboring the human and bovine LDH genes was higher than that of the strains harboring lactic acid bacteria LDH genes. DNA microarray analysis revealed that the expression of 388 genes was significantly altered in the strains with the human and bovine LDH genes. Of these, the L-lactate productivity of human LDH-harboring deletion strains of 289 genes was compared with that of the standard and 56 randomly selected deletion strains containing the same LDH gene to validate the effectiveness of DNA microarray analysis for identifying the genes responsible for L-lactate production in the recombinant strains. Only deletion strains of the genes selected on the basis of the DNA microarray data showed significantly altered L-lactate production as compared to the standard and the randomly selected deletion strains. Our results indicated that the genes related to L-lactate production could be successfully identified by selecting the genes that exhibited significantly altered expression on DNA microarray analysis, and the effectiveness of DNA microarray analysis for identifying the genes responsible for L-lactate production was discussed.
为了确定通过DNA微阵列分析获得的转录组数据是否可用于鉴定参与目标代谢物产生的基因，我们试图鉴定产L-乳酸的重组酿酒酵母菌株中参与L-乳酸产生的基因。我们获得了这些菌株的DNA微阵列数据。将携带乳酸菌、牛和人L-乳酸脱氢酶（LDH）基因的质粒导入PDC1基因缺失的酿酒酵母菌株中。携带人和牛LDH基因的菌株的L-乳酸生产率高于携带乳酸菌LDH基因的菌株。DNA微阵列分析显示，在携带人和牛LDH基因的菌株中，388个基因的表达发生了显著变化。其中，将289个基因的人源LDH缺失菌株的L-乳酸生产率与标准菌株以及56个随机选择的含有相同LDH基因的缺失菌株进行比较，以验证DNA微阵列分析在鉴定重组菌株中负责L-乳酸产生的基因方面的有效性。与标准菌株和随机选择的缺失菌株相比，只有基于DNA微阵列数据选择的基因的缺失菌株显示出L-乳酸产量有显著变化。我们的结果表明，通过选择在DNA微阵列分析中表达显著改变的基因，可以成功鉴定与L-乳酸产生相关的基因，并讨论了DNA微阵列分析在鉴定负责L-乳酸产生的基因方面的有效性。
