title: In situ immobilized lipase on the surface of intracellular polyhydroxybutyrate granules: preparation, characterization, and its promising use for the synthesis of fatty acid alkyl esters.
abstract: Photobacterium lipolyticum M37 lipase (LipM37) was immobilized on the surface of intracellular polyhydroxybutyrate (PHB) granules in Escherichia coli. LipM37 was genetically fused to Cupriavidus necator PHA synthase (PhaC Cn ), and the engineered PHB operon containing the lip M37 -phaC Cn successfully mediated the accumulation of PHB granules (85 wt.%) inside E. coli cells. The PHB granules were isolated from the crude cell extract, and the immobilized LipM37 was comparable with the free form of LipM37 except for a favorable increase in thermostability. The immobilized LipM37 was used to synthesize oleic acid methyl ester (biodiesel) and oleic acid dodecyl ester (wax ester), and yielded 98.0 % conversion in esterification of oleic acid and dodecanol. It was suggested that the LipM37-PhaCCn fusion protein successfully exhibited bifunctional activities in E. coli and that in situ immobilization of lipase to the intracellular PHB could be a promising approach for expanding the biocatalytic toolbox for industrial chemical synthesis.
authors.affiliation: R&D Center, Genofocus Inc., 187 Techno-ro, Yuseong-gu, Daejeon, 305-500, Republic of Korea.
authors.forename: Taek Ho
authors.initials: TH
authors.lastname: Yang
authors.affiliation: Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology (KRICT), 141 Gajeong-ro, Yuseong-gu, Daejeon, 305-600, Republic of Korea.
authors.forename: Min-A
authors.initials: MA
authors.lastname: Kwon
authors.affiliation: Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology (KRICT), 141 Gajeong-ro, Yuseong-gu, Daejeon, 305-600, Republic of Korea.
authors.forename: Ji Young
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology (KRICT), 141 Gajeong-ro, Yuseong-gu, Daejeon, 305-600, Republic of Korea.
authors.forename: Ji-Eun
authors.initials: JE
authors.lastname: Choi
authors.affiliation: Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology (KRICT), 141 Gajeong-ro, Yuseong-gu, Daejeon, 305-600, Republic of Korea.
authors.forename: Joon Young
authors.initials: JY
authors.lastname: Oh
authors.affiliation: Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology (KRICT), 141 Gajeong-ro, Yuseong-gu, Daejeon, 305-600, Republic of Korea. ajee@krict.re.kr.
authors.forename: Jae Kwang
authors.initials: JK
authors.lastname: Song
mesh_terms: D055162:Biocatalysis; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism*; D004926:Escherichia coli / Q000166:cytology; D004951:Esterification; D004952:Esters; D006885:Hydroxybutyrates / Q000737:chemistry* / Q000378:metabolism*; D042541:Intracellular Space / Q000737:chemistry*; D008049:Lipase / Q000737:chemistry* / Q000378:metabolism*; D010776:Photobacterium / Q000201:enzymology; D011091:Polyesters / Q000737:chemistry* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013499:Surface Properties; D000094122:Polyhydroxybutyrates
chemical_list: D004800:Enzymes, Immobilized; D004952:Esters; D006885:Hydroxybutyrates; D008049:Lipase; D011091:Polyesters; D011993:Recombinant Fusion Proteins; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates
keywords: Esterification; Fatty acid alkyl ester; In situ immobilization; Lipase; Polyhydroxybutyrate