title: Overexpression of malic enzyme (ME) of Mucor circinelloides improved lipid accumulation in engineered Rhodotorula glutinis.
abstract: The oleaginous yeast Rhodotorula glutinis has been known to be a potential feedstock for lipid production. In the present study, we investigated the enhancement of expression of malic enzyme (ME; NADP(+) dependent; EC 1.1.1.40) from Mucor circinelloides as a strategy to improve lipid content inside the yeast cells. The 26S rDNA and 5.8S rDNA gene fragments isolated from Rhodotorula glutinis were used for homologous integration of ME gene into R. glutinis chromosome under the control of the constitutively highly expressed gene phosphoglycerate kinase 1 to achieve stable expression. We demonstrated that by increasing the expression of the foreign ME gene in R. glutinis, we successfully improved the lipid content by more than twofold. At the end of lipid accumulation phrase (96 h) in the transformants, activity of ME was increased by twofold and lipid content of the yeast cells was increased from 18.74 % of the biomass to 39.35 %. Simultaneously, there were no significant differences in fatty acid profiles between the wild-type strain and the recombinant strain. Over 94 % of total fatty acids were C16:0, C18:0, C16:1, C18:1, and C18:2. Our results indicated that heterologous expression of NADP(+)-dependent ME involved in fatty acid biosynthesis indeed increased the lipid accumulation in the oleaginous yeast R. glutinis.
authors.affiliation: Institute of Genomic Medicine, College of Pharmacy, Jinan University, Guangzhou 510632, China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Li
authors.forename: Hanxiao
authors.initials: H
authors.lastname: Sun
authors.forename: Xuemei
authors.initials: X
authors.lastname: Mo
authors.forename: Xiuying
authors.initials: X
authors.lastname: Li
authors.forename: Bo
authors.initials: B
authors.lastname: Xu
authors.forename: Peng
authors.initials: P
authors.lastname: Tian
mesh_terms: D003067:Coenzymes / Q000378:metabolism; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D005227:Fatty Acids / Q000378:metabolism*; D015870:Gene Expression; D005818:Genetic Engineering; D008969:Molecular Sequence Data; D009089:Mucor / Q000201:enzymology* / Q000235:genetics; D009249:NADP / Q000378:metabolism; D012335:RNA, Ribosomal / Q000235:genetics; D012340:RNA, Ribosomal, 5.8S / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012248:Rhodotorula / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA
chemical_list: D003067:Coenzymes; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D005227:Fatty Acids; D012335:RNA, Ribosomal; D012340:RNA, Ribosomal, 5.8S; C065096:RNA, ribosomal, 26S; D011994:Recombinant Proteins; D009249:NADP