Subject Area物理化学
Cloning and characterization of the isocitrate dehydrogenase gene from oleaginous yeast Lipomyces starkeyi
Tang W(唐伟); Zhang SF(张素芳); Tan HD(谭海东); Zhao ZB(赵宗保)
Conference Name13th International Biotechnology Symposium and Exhibition
Conference Date2008-10-12
Conference Place中国
Funding Organization中国科学院,中国工程院,中国教育部,中国科技部
AbstractOleaginous yeast Lipomyces starkeyi can accumulate intracellular lipids over 65% of its cell dry weight [1]. Mitochondrial NAD+-specific isocitrate dehydrogenase (IDH) catalyzes the oxidative decarboxylation of isocitrare to α-ketoglutarate in eukaryotic cells, an essential and rate-limiting step in citric acid cycle. Yeast S. cerevisiae IDH is composed of an octamer of four IDH1 and four IDH2 subunits [2]. Early biochemical study showed that IDH activity correlates with the lipid storage capacity [3]. In this work we cloned the full length cDNA sequence of the mitochondrial IDH1 and IDH2 gene using methods of degenerate polymerase chain reaction (PCR) and rapid amplification of cDNA ends from L. starkeyi AS 2.1560, a useful oil-producing yeast. We further determined the expression level of both lsidh1 and lsidh2 by real time PCR during the lipid storage process. Our results showed the encoded products, LsIDH1 and LsIDH2, are similar in size with a molecular weight of 40,277 and 40,708, respectively, and have high similarity with IDH sequences from other yeast species. More importantly, putative mitochondrial transit peptides have been identified at the N-terminus of each sequence, suggesting that both proteins are likely located within the mitochondria. Further gene expression analysis by real time PCR showed that both idh1 and idh2 expression decreased concurrently with the evolution of cellular lipids. Our data may be valuable for further engineering oleaginous yeasts.
WOS IDWOS:000208979401333
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Document Type会议论文
Corresponding AuthorZhao ZB(赵宗保)
Recommended Citation
GB/T 7714
Tang W,Zhang SF,Tan HD,et al. Cloning and characterization of the isocitrate dehydrogenase gene from oleaginous yeast Lipomyces starkeyi[C],2008:S359/2.
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