1. Identification and characterization of a flavin-containing monooxygenase MoA and its function in a specific sophorolipid molecule metabolism in Starmerella bombicola.
- Author
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Li, Jiashan, Li, Hui, Li, Weiwei, Xia, Chengqiang, and Song, Xin
- Subjects
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MONOOXYGENASES , *FLAVINS , *FUNGAL metabolism , *GENOMICS , *DELETION mutation , *FUNGAL mutation - Abstract
The yeast Starmerella bombicola CGMCC 1576 can produce abundant sophorolipids (SLs) including almost equal proportion of acidic and lactonic SLs. In this study, a monooxygenase MoA responsible for the metabolism of a sophorolipid molecule, C18:2 diacetylated acidic sophorolipid (C18:2 DASL), was identified, through genomic analysis, protein modeling, and gene knocking out strategy. The yield and compositions of SLs produced by the deletion mutant ∆moA changed dramatically. In HPLC chromatogram, the UV absorption area of C18:2 DASL (one major acidic sophorolipid component) increased from 9.84 × 10 mAU × s to 34.26 × 10 mAU × s by an increase of 248.17 % when oleic acid was used as hydrophobic carbon source. Moreover, when linoleic acid was used as hydrophobic carbon source, the content of C18:2 DASL component produced by the overexpressed strain Peno::moA decreased significantly compared with that of wild type and △ moA. Furthermore, the MoA enzyme was heterologously expressed in Escherichia coli JM109 (DE3) with a recombinant plasmid named pMAL-c2x-moA, and the purified enzyme was obtained through a maltose-binding protein (MBP) affinity chromatography column. The purified C18:2 DASL and C18:1 DASL were applied to be catalyzed by MoA enzyme, respectively; it turned to be that C18:1 DASL still remained in the MoA reaction system, but C18:2 DASL disappeared. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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