Cloning of delta6-desaturase from Mucor circinelloides and its high expression in Saccharomyces cerevisiae.
The aim of this study is to obtain Saccharomyces cerevisiae engineering strain with high gamma-linolenic acid (GLA, gamma-C18:3), which is a nutritionally important fatty acid that plays a vital role in biological structure and cell functions. As the first step,we cloned gamma6-desaturase gene from fungus mucor circinelloides by RT-PCR; delta6-desaturase is responsible for the transformation of linoleic acid into GLA. The PCR product was subcloned into yeast expression vector pYES2 to generate a recombinant plasmid pYES412. Transformation of S. cerevisiae strain INVSc1 was done by the lithium acetate method and the recombinant yeast cells were selected on a uracil-deficient medium. On appropriate medium and temperature,linoleic acid was provided as a substrate to yeast cultures,and the level of gamma-linolenic acid reached 50.07%. So far,the result we obtained is the best in terms of the level of expression of delta6-desaturase gene in Saccharomyces cerevisiae.
Hao
Yan-Ling
YL
Department of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China. yanlinghao@163.com
Wang
Ying
Y
Zhu
Ben-Zhong
BZ
Luan
Chun-Guang
CG
Luo
Yun-Bo
YB
D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D050602:Linoleoyl-CoA Desaturase / Q000235:genetics / Q000378:metabolism*; D009089:Mucor / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013696:Temperature; D017965:gamma-Linolenic Acid / Q000378:metabolism
D018076:DNA, Complementary; D005656:Fungal Proteins; D011994:Recombinant Proteins; D017965:gamma-Linolenic Acid; D050602:Linoleoyl-CoA Desaturase