Dalian Institute of Chemical Physics CAS, China.
Can J Microbiol. 2009 Sep;55(9):1062-9. doi: 10.1139/w09-063.
The oleaginous yeast Lipomyces starkeyi can accumulate intracellular lipids to over 60% of its cell dry mass under a nitrogen-limited condition. We showed that extracellular and intracellular citrate concentrations of L. starkeyi AS 2.1560 increased and the nicotinamide adenine dinucleotide - isocitrate dehydrogenase (NAD+-IDH) activity decreased at the beginning of the lipid accumulation, suggesting that the attenuation of the NAD+-IDH activity might initiate lipid storage. We next cloned the IDH gene by the methods of degenerate PCR and rapid amplification of cDNA ends. Phylogenetic analyses of the evolutionary relationships among LsIDH1, LsIDH2, and other yeast NAD+-IDHs revealed that the L. starkeyi IDH had a closer relationship with the IDHs of Yarrowia lipolytica. Further real-time PCR analysis showed that the expression levels of both LsIDH1 and LsIDH2 decreased concurrently with the evolution of cellular lipids. Our data should be valuable for understanding the biology of oleaginous yeasts and for further strain engineering of L. starkeyi.
产油酵母拉斯克氏油脂酵母在氮限制条件下可以将细胞内的脂质积累到超过其细胞干重的 60%。我们发现,在油脂积累的初期,拉斯克氏油脂酵母 AS 2.1560 的细胞外和细胞内柠檬酸浓度增加,烟酰胺腺嘌呤二核苷酸 - 异柠檬酸脱氢酶(NAD+-IDH)活性降低,表明 NAD+-IDH 活性的衰减可能引发脂质储存。接下来,我们通过简并 PCR 和快速扩增 cDNA 末端的方法克隆了 IDH 基因。LsIDH1、LsIDH2 和其他酵母 NAD+-IDHs 之间的进化关系的系统发育分析表明,拉斯克氏油脂酵母的 IDH 与解脂耶氏酵母的 IDH 关系更密切。进一步的实时 PCR 分析表明,随着细胞内脂质的积累,LsIDH1 和 LsIDH2 的表达水平同时降低。我们的数据对于理解产油酵母的生物学和进一步对拉斯克氏油脂酵母进行菌株工程改造应该是有价值的。