title: Identification and characterization of a long-chain fatty acid transporter in the sophorolipid-producing strain Starmerella bombicola.
abstract: The sophorolipid-producing strain Starmerella bombicola CGMCC 1576 has a remarkable ability to produce sophorolipids (SLs) under the acidic and lactonic forms with almost equal proportion. In this study, we found the gene encoding for the long-chain acyl-CoA synthetase (ALCS). This enzyme was putatively identified as a membrane-bound long-chain fatty acid transport protein and contributed to the uptake of long-chain fatty acids. Disruption of the alcs gene resulted in an impaired growth of the alcs-deleted mutant in minimal media containing different fatty acids (C12:0, C14:0, C16:0, C18:0, C22:0, and C24:0) as the sole carbon source and led to a dramatic decrease in the uptake of the fluorescent-tagged long-chain fatty acid analogue-boron dipyrromethene difluoride dodecanoic acid (BODIPY-3823). The absence of this alcs gene caused obvious phenotype changes. Compared with the wild-type strain, the yield and compositions of the SLs produced by the gene-deleted mutant of ∆alcs::six showed almost no lactonic form of SLs, and the acidic SLs were composed of medium-chain. The ALCS enzyme was heterologously expressed in Escherichia coli JM109 (DE3) with pMAL-c2x-alcs. The enzyme was purified through a maltose-binding protein (MBP) affinity chromatography column and was confirmed to be homogeneous by SDS-PAGE. The recombinant enzyme could catalyze the formation of the long-chain acyl-CoA when the long-chain fatty acids and the coenzyme A were used as substrates.
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Shan Da Nan Road 27, Jinan, Shandong, 250100, People's Republic of China.
authors.forename: Jiashan
authors.initials: J
authors.lastname: Li
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Shan Da Nan Road 27, Jinan, Shandong, 250100, People's Republic of China.
authors.forename: Chengqiang
authors.initials: C
authors.lastname: Xia
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Shan Da Nan Road 27, Jinan, Shandong, 250100, People's Republic of China.
authors.forename: Xiaoran
authors.initials: X
authors.lastname: Fang
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Shan Da Nan Road 27, Jinan, Shandong, 250100, People's Republic of China.
authors.forename: Haizhao
authors.initials: H
authors.lastname: Xue
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Shan Da Nan Road 27, Jinan, Shandong, 250100, People's Republic of China. songx@sdu.edu.cn.|National Glycoengineering Research Center, Shandong University, Shan Da Nan Road 27, Jinan, Shandong, 250100, People's Republic of China. songx@sdu.edu.cn.
authors.forename: Xin
authors.initials: X
authors.lastname: Song
mesh_terms: D000595:Amino Acid Sequence; D001692:Biological Transport / Q000235:genetics; D003066:Coenzyme A Ligases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D050612:Fatty Acid Transport Proteins / Q000235:genetics*; D005227:Fatty Acids / Q000378:metabolism*; D006017:Glycolipids / Q000096:biosynthesis*; D058619:Maltose-Binding Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D004718:Saccharomycetales / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D016415:Sequence Alignment; D000098859:Long-Chain-Fatty-Acid-CoA Ligase
chemical_list: D050612:Fatty Acid Transport Proteins; D005227:Fatty Acids; D006017:Glycolipids; D058619:Maltose-Binding Proteins; D011994:Recombinant Proteins; D003066:Coenzyme A Ligases; D000098859:Long-Chain-Fatty-Acid-CoA Ligase
keywords: Acyl-CoA synthetase; Fatty acid uptake; Sophorolipids; Starmerella bombicola