caa a22 4500 605506051 CHVBK 20210128100625.0 cr unu---uuuuu 210128e20150701xx s 000 0 eng 10.1007/s00253-014-6353-8 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6353-8 Characterization of an omega-6 linoleate lipoxygenase from Burkholderia thailandensis and its application in the production of 13-hydroxyoctadecadienoic acid [Elektronische Daten] [Jung-Ung An, Baek-Joong Kim, Seung-Hye Hong, Deok-Kun Oh] A recombinant putative lipoxygenase from Burkholderia thailandensis with a specific activity of 26.4Umg−1 was purified using HisTrap affinity chromatography. The native enzyme was a 75-kDa dimer with a molecular mass of 150kDa. The enzyme activity and catalytic efficiency (k cat/K m) were the highest for linoleic acid (k cat of 93.7s−1 and K m of 41.5μM), followed by arachidonic acid, α-linolenic acid, and γ-linolenic acid. The enzyme was identified as an omega-6 linoleate lipoxygenase (or a linoleate 13S-lipoxygenase) based on genetic and HPLC analyses as well as substrate specificity. The reaction conditions for the enzymatic production of 13-hydroxy-9,11(Z,E)-octadecadienoic acid (13-HODE) were optimal at pH 7.5, 25°C, 20gl−1 linoleic acid, 2.5gl−1 enzyme, 0.1mM Cu2+, and 6% (v/v) methanol. Under these conditions, linoleate 13-lipoxygenase from B. thailandensis produced 20.8gl−1 13-HODE (70.2mM) from 20gl−1 linoleic acid (71.3mM) for 120min, with a molar conversion yield of 98.5% and productivity of 10.4gl−1h−1. The molar conversion yield and productivity of 13-HODE obtained using B. thailandensis lipoxygenase were 151 and 158% higher, respectively, than those obtained using commercial soybean lipoxygenase under the optimum conditions for each enzyme at the same concentrations of substrate and enzyme. Springer-Verlag Berlin Heidelberg, 2015 Burkholderia thailandensis nationallicence 13-Hydroxyoctadecadienoic acid nationallicence 13-Hydroperoxyctadecadienoic acid nationallicence Linoleate 13-Lipoxygenase nationallicence Linoleic acid nationallicence An Jung-Ung Department of Bioscience and Biotechnology, Konkuk University, 143-701, Seoul, Republic of Korea aut Kim Baek-Joong Department of Bioscience and Biotechnology, Konkuk University, 143-701, Seoul, Republic of Korea aut Hong Seung-Hye Department of Bioscience and Biotechnology, Konkuk University, 143-701, Seoul, Republic of Korea aut Oh Deok-Kun Department of Bioscience and Biotechnology, Konkuk University, 143-701, Seoul, Republic of Korea aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/13(2015-07-01), 5487-5497 0175-7598 99:13<5487 2015 99 253 https://doi.org/10.1007/s00253-014-6353-8 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6353-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- An Jung-Ung Department of Bioscience and Biotechnology, Konkuk University, 143-701, Seoul, Republic of Korea aut NATIONALLICENCE 700 1- Kim Baek-Joong Department of Bioscience and Biotechnology, Konkuk University, 143-701, Seoul, Republic of Korea aut NATIONALLICENCE 700 1- Hong Seung-Hye Department of Bioscience and Biotechnology, Konkuk University, 143-701, Seoul, Republic of Korea aut NATIONALLICENCE 700 1- Oh Deok-Kun Department of Bioscience and Biotechnology, Konkuk University, 143-701, Seoul, Republic of Korea aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/13(2015-07-01), 5487-5497 0175-7598 99:13<5487 2015 99 253