期刊
GENE
卷 706, 期 -, 页码 106-114出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.gene.2019.04.074
关键词
Aspergillus oryzae; Diacylglycerol acyltransferase; Dihomo-gamma-linolenic acid; Lipid accumulation; Triacylglycerol; Oleaginous fungi
资金
- Chalermphrakiat Grant, Faculty of Medicine Siriraj Hospital, Mahidol University
- Center of Excellence for Innovation in Chemistry (PERCH-CIC), the Office of the Higher Education Commission
- National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand [P15-51082]
Biological significance of 18-carbon polyunsaturated fatty acids, gamma-linolenic acid (GLA; C18:3 n-6) and dihomo-gamma-linolenic acid (DGLA; C20:3 n-6) has gained much attention in the systematic development of optimized strains for industrial applications. In this work, a n-6 PUFAs-producing strain of Aspergillus oryzae was generated by manipulating metabolic reactions in fatty acid modification and triacylglycerol biosynthesis. The codon-optimized genes coding for Delta(6)-desaturase and Delta(6)-elongase of Pythium sp., and diacylglycerol acyltransferase 2 (mMaDGAT2) of Mortierella alpina were co-transformed in a single vector into A. oryzae BCC14614, yielding strain TD6E6-DGAT2. Comparative phenotypic analysis showed that a 70% increase of lipid titer was found in the engineered strain, which was a result of a significant increase in triacylglycerol (TAG) content (52.0 +/- 1.8% of total lipids), and corresponded to the increased size of lipid particles observed in the fungal cells. Interestingly, the proportions of GLA and DGLA in neutral lipids of the engineered strain were similar, with the highest titers obtained in the high C:N culture (29:0; 6% glucose) during the lipid-accumulating stage of growth. Time-course expression analysis of the engineered strain revealed transcriptional control of TAG biosynthesis through a co-operation between the native DGAT2 of A. oryzae and the transformed mMaDGAT2.
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