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Characterization and robust nature of newly isolated oleaginous marine yeast Rhodosporidium spp. from coastal water of Northern China.

ABSTRACT: A total of ten marine yeast strains isolated from Bohai Sea, Northern China were identified to be members of three genera Rhodosporidium, Rhodotorula, and Cryptococcus. Two representative strains Rhodosporidium TJUWZ4 and Cryptococcus TJUWZA11 with high lipid content based on Nile red staining method were further characterized. A wide range of culture conditions (C and N sources, pH, temperature, salinity and C/N ratio) were tested to characterize the biomass and lipid production (yield and productivity) of these strains. Results indicated that Rhodosporidium TJUWZ4 was capable of achieving lipid yield up to 44% and 0.09 g/l-h productivity on glucose and peptone medium at pH 4, 20 °C, 30% salinity, and C/N 80. Three fatty acids, namely oleic acid (18:1), palmitic acid (C16:0) and linoleic acid (18:2) were the major intracellular fatty acids, which accounted for 90% of total lipids. With promising features for intracellular lipid accumulation, Rhodosporidium TJUWZ4 is a robust strain with great potentials for application in biodiesel production from renewable feedstocks.

SUBMITTER: Wang Q

PROVIDER: S-EPMC5302000 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature

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Characterization and robust nature of newly isolated oleaginous marine yeast Rhodosporidium spp. from coastal water of Northern China.

Wang Qiuzhen Q Cui Yan Y Sen Biswarup B Ma Wenmeng W Zheng Rose Lynn RL Liu Xianhua X Wang Guangyi G

AMB Express 20170201 1

A total of ten marine yeast strains isolated from Bohai Sea, Northern China were identified to be members of three genera Rhodosporidium, Rhodotorula, and Cryptococcus. Two representative strains Rhodosporidium TJUWZ4 and Cryptococcus TJUWZA11 with high lipid content based on Nile red staining method were further characterized. A wide range of culture conditions (C and N sources, pH, temperature, salinity and C/N ratio) were tested to characterize the biomass and lipid production (yield and prod ...[more]

PMID: 28144888

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Project description:BackgroundRed yeast species in the Rhodotorula/Rhodosporidium genus are outstanding producers of triacylglyceride and cell biomass. Metabolic engineering is expected to further enhance the productivity and versatility of these hosts for the production of biobased chemicals and fuels. Promoters with strong activity during oil-accumulation stage are critical tools for metabolic engineering of these oleaginous yeasts.ResultsThe upstream DNA sequences of 6 genes involved in lipid biosynthesis or accumulation in Rhodotorula toruloides were studied by luciferase reporter assay. The promoter of perilipin/lipid droplet protein 1 gene (LDP1) displayed much stronger activity (4-11 folds) than that of glyceraldehyde-3-phosphate dehydrogenase gene (GPD1), one of the strongest promoters known in yeasts. Depending on the stage of cultivation, promoter of acetyl-CoA carboxylase gene (ACC1) and fatty acid synthase β subunit gene (FAS1) exhibited intermediate strength, displaying 50-160 and 20-90% levels of GPD1 promoter, respectively. Interestingly, introns significantly modulated promoter strength at high frequency. The incorporation of intron 1 and 2 of LDP1 (LDP1in promoter) enhanced its promoter activity by 1.6-3.0 folds. Similarly, the strength of ACC1 promoter was enhanced by 1.5-3.2 folds if containing intron 1. The intron 1 sequences of ACL1 and FAS1 also played significant regulatory roles. When driven by the intronic promoters of ACC1 and LDP1 (ACC1in and LDP1in promoter, respectively), the reporter gene expression were up-regulated by nitrogen starvation, independent of de novo oil biosynthesis and accumulation. As a proof of principle, overexpression of the endogenous acyl-CoA-dependent diacylglycerol acyltransferase 1 gene (DGA1) by LDP1in promoter was significantly more efficient than GPD1 promoter in enhancing lipid accumulation.ConclusionIntronic sequences play an important role in regulating gene expression in R. toruloides. Three intronic promoters, LDP1in, ACC1in and FAS1in, are excellent promoters for metabolic engineering in the oleaginous and carotenogenic yeast, R. toruloides.

Dataset Information

Characterization and robust nature of newly isolated oleaginous marine yeast Rhodosporidium spp. from coastal water of Northern China.

Publications

Characterization and robust nature of newly isolated oleaginous marine yeast Rhodosporidium spp. from coastal water of Northern China.

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