Abstract
A novel expression system was established in the oleaginous yeast, Lipomyces kononenkoae. The expression vector pLK-rhPHG of L. kononenkoae was constructed and using the hygromycin phosphotransferase gene and green fluorescent protein gene as reporter genes. A delta 6-fatty acid desaturase gene (D6DM) from Cunninghamella echinulata MIAN6 was then expressed in this strain. The recombinant strain accumulated about 1.2% γ-linolenic acid in the total fatty acids.
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Acknowledgments
This work was supported by National Natural Science Foundation of China (Grants 31000778 and 30970262) and the Major S&T Projects on the Cultivation of New Varieties of Genetically Modified Organisms (Grant 2009ZX08009-120B) from Ministry of Agriculture, P.R. China.
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Ping Wang and Xia Wan contributed equally to this work.
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10529_2011_651_MOESM2_ESM.tif
Supplementary Fig. 2 Recovery of pLK-rhPHG-transformed L. kononenkoae. A, L. kononenkoae harboring pLK-rhPHG grown on a YPD plate supplemented with 60 mg hygromycin B/l; B, L. kononenkoae grown on a YPD plate supplemented with 60 mg hygromycin B/l (TIFF 830 kb)
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Supplementary Fig. 3 Expression of the GFP reporter gene in L. kononenkoae. A, Light microscopy of L. kononenkoae; B, fluorescence microscopy of L. kononenkoae; C, light microscopy of L. kononenkoae harboring pLK-rhPHG; D, fluorescence microscopy of L. kononenkoae harboring pLK-rhPHG (TIFF 968 kb)
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Supplementary Fig. 4 Gas chromatography analysis of fatty acid compositions A, GLA standard, B, L. kononenkoae harboring pLK-rhPHT C, L. kononenkoae 1714 harboring pLK-rhPHM (TIFF 879 kb)
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Wang, P., Wan, X., Zhang, Y. et al. Production of γ-linolenic acid using a novel heterologous expression system in the oleaginous yeast Lipomyces kononenkoae . Biotechnol Lett 33, 1993–1998 (2011). https://doi.org/10.1007/s10529-011-0651-3
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DOI: https://doi.org/10.1007/s10529-011-0651-3