title: Enhanced production of polyunsaturated fatty acids by enzyme engineering of tandem acyl carrier proteins.
abstract: In some microorganisms, polyunsaturated fatty acids (PUFAs) are biosynthesized by PUFA synthases characterized by tandem acyl carrier proteins (ACPs) in subunit A. These ACPs were previously shown to be important for PUFA productivity. In this study, we examined their function in more detail. PUFA productivities increased depending on the number of ACPs without profile changes in each subunit A of eukaryotic and prokaryotic PUFA synthases. We also constructed derivative enzymes from subunit A with 5 × ACPs. Enzymes possessing one inactive ACP at any position produced ~30% PUFAs compared with the parental enzyme but unexpectedly had ~250% productivity compared with subunit A with 4 × ACPs. Enzymes constructed by replacing the 3rdACP with an inactive ACP from another subunit A or ACP-unrelated sequences produced ~100% and ~3% PUFAs compared with the parental 3rdACP-inactive enzyme, respectively. These results suggest that both the structure and number of ACP domains are important for PUFA productivity.
authors.affiliation: Graduate School of Chemical Sciences and Engineering, Hokkaido University, N13-W8, Kita-ku, Sapporo 060-8628, Japan.
authors.forename: Shohei
authors.initials: S
authors.lastname: Hayashi
authors.affiliation: Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, Sapporo 060-8628, Japan.
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Satoh
authors.affiliation: Kyowa Hakko Bio Co. Ltd., 1-6-1, Ohtemachi, Chiyoda-ku, Tokyo 100-8185, Japan.
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Ujihara
authors.affiliation: Research Faculty of Agriculture, Hokkaido University, N9-W9, Kita-ku, Sapporo 060-8589, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Takata
authors.affiliation: Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, Sapporo 060-8628, Japan.
authors.forename: Tohru
authors.initials: T
authors.lastname: Dairi
mesh_terms: D000213:Acyl Carrier Protein / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D053898:Biosynthetic Pathways; D005231:Fatty Acids, Unsaturated / Q000096:biosynthesis*; D000072417:Protein Domains; D015202:Protein Engineering / Q000379:methods
chemical_list: D000213:Acyl Carrier Protein; D005231:Fatty Acids, Unsaturated