title: Unveiling of novel regio-selective fatty acid double bond hydratases from Lactobacillus acidophilus involved in the selective oxyfunctionalization of mono- and di-hydroxy fatty acids.
abstract: The aim of this study is the first time demonstration of cis-12 regio-selective linoleate double-bond hydratase. Hydroxylation of fatty acids, abundant feedstock in nature, is an emerging alternative route for many petroleum replaceable products thorough hydroxy fatty acids, carboxylic acids, and lactones. However, chemical route for selective hydroxylation is still quite challenging owing to low selectivity and many environmental concerns. Hydroxylation of fatty acids by hydroxy fatty acid forming enzymes is an important route for selective biocatalytic oxyfunctionalization of fatty acids. Therefore, novel fatty acid hydroxylation enzymes should be discovered. The two hydratase genes of Lactobacillus acidophilus were identified by genomic analysis, and the expressed two recombinant hydratases were identified as cis-9 and cis-12 double-bond selective linoleate hydratases by in vitro functional validation, including the identification of products and the determination of regio-selectivity, substrate specificity, and kinetic parameters. The two different linoleate hydratases were the involved enzymes in the 10,13-dihydroxyoctadecanoic acid biosynthesis. Linoleate 13-hydratase (LHT-13) selectively converted 10 mM linoleic acid to 13S-hydroxy-9(Z)-octadecenoic acid with high titer (8.1 mM) and yield (81%). Our study will expand knowledge for microbial fatty acid-hydroxylation enzymes and facilitate the designed production of the regio-selective hydroxy fatty acids for useful chemicals from polyunsaturated fatty acid feedstocks.
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, 143-701, Republic of Korea.
authors.forename: Kyoung-Rok
authors.initials: KR
authors.lastname: Kim
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, 143-701, Republic of Korea.
authors.forename: Hye-Jin
authors.initials: HJ
authors.lastname: Oh
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, 143-701, Republic of Korea.
authors.forename: Chul-Soon
authors.initials: CS
authors.lastname: Park
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, 143-701, Republic of Korea.
authors.forename: Seung-Hye
authors.initials: SH
authors.lastname: Hong
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, 143-701, Republic of Korea.
authors.forename: Ji-Young
authors.initials: JY
authors.lastname: Park
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, 143-701, Republic of Korea. deokkun@konkuk.ac.kr.
authors.forename: Deok-Kun
authors.initials: DK
authors.lastname: Oh
mesh_terms: D005227:Fatty Acids / Q000378:metabolism*; D006836:Hydro-Lyases / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D007779:Lactobacillus acidophilus / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D005227:Fatty Acids; D011994:Recombinant Proteins; D006836:Hydro-Lyases
keywords: 10,13-dihydroxy-octadecanoic acid; 13-hydroxy-9(Z)-octadecenoic acid; Lactobacillus acidophilus; hydroxy fatty acid; linoleate 13-hydratase; linoleic acid; regio-selective hydroxylation