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Characterization of an omega-6 linoleate lipoxygenase from Burkholderia thailandensis and its application in the production of 13-hydroxyoctadecadienoic acid

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A recombinant putative lipoxygenase from Burkholderia thailandensis with a specific activity of 26.4 U mg−1 was purified using HisTrap affinity chromatography. The native enzyme was a 75-kDa dimer with a molecular mass of 150 kDa. The enzyme activity and catalytic efficiency (k cat/K m) were the highest for linoleic acid (k cat of 93.7 s−1 and K m of 41.5 μM), 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, 20 g l−1 linoleic acid, 2.5 g l−1 enzyme, 0.1 mM Cu2+, and 6 % (v/v) methanol. Under these conditions, linoleate 13-lipoxygenase from B. thailandensis produced 20.8 g l−1 13-HODE (70.2 mM) from 20 g l−1 linoleic acid (71.3 mM) for 120 min, with a molar conversion yield of 98.5 % and productivity of 10.4 g l−1 h−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.

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This study was supported by grants from the Bio-industry Technology Development Program, Ministry for Agriculture, Food and Rural Affairs (No. 112002-3) and the Korea Healthcare Technology R&D Project, Ministry for Health & Welfare, Republic of Korea (No. 2012-009).

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Correspondence to Deok-Kun Oh.

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An, JU., Kim, BJ., Hong, SH. et al. Characterization of an omega-6 linoleate lipoxygenase from Burkholderia thailandensis and its application in the production of 13-hydroxyoctadecadienoic acid. Appl Microbiol Biotechnol 99, 5487–5497 (2015).

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