Catalytic characterization of heterodimeric linoleate 13S-lipoxygenase from black soybean (Glycine max (L.) Merr.),Enzyme and Microbial Technology

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Catalytic characterization of heterodimeric linoleate 13S-lipoxygenase from black soybean (Glycine max (L.) Merr.)
Enzyme and Microbial Technology ( IF 3.4 ) Pub Date : 2020-09-01 , DOI: 10.1016/j.enzmictec.2020.109595
Jun-Young Park ₁ , Chae Hyung Kim ₁ , Yoonseok Choi ₁ , Kyung-Min Park ₂ , Pahn-Shick Chang ₃

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A novel lipoxygenase (BLOX) was purified from black soybean (Glycine max (L.) Merr.), and its catalytic properties were characterized. The molecular weight of BLOX was 101 kDa and its unique heterodimeric structure with two different subunits of molecular weight 46 kDa and 55 kDa was elucidated. The optimum pH and temperature of BLOX were pH 9.5 and 40 °C, respectively. BLOX was highly stable at the pH range of 6.0-10.0 and below 40 °C, and was stimulated by adding ferrous ion (Fe2+). In terms of substrate specificity, BLOX showed a substrate preference to linoleic acid that is the main substance to produce hydroperoxides in soybean. When it reacted with linoleic acid, the major product was 13(S)-hydroperoxy-9,11-octadecadienoic acid; therefore, it could be classified into the linoleate 13S-LOX family (EC 1.13.11.12). Finally, the kinetic parameters (Vmax, Km, and kcat) of BLOX were 0.124 mM min-1, 0.636 mM, and 12.28 s-1, respectively, and consequently, the catalytic efficiency (kcat/Km) was calculated as 1.93 × 104 M-1·s-1. These catalytic characteristics of BLOX could contribute to understanding the enzymatic rancidification of black soybean, and to further biotechnical approaches to control and mitigate the deterioration.

中文翻译：

黑大豆 (Glycine max (L.) Merr.) 异二聚亚油酸 13S-脂氧合酶的催化特性

从黑大豆 (Glycine max (L.) Merr.) 中纯化出一种新型脂氧合酶 (BLOX)，并对其催化性能进行了表征。 BLOX 的分子量为 101 kDa，其独特的异二聚体结构具有分子量 46 kDa 和 55 kDa 两个不同的亚基。 BLOX 的最适pH 和温度分别为pH 9.5 和40 °C。 BLOX 在 6.0-10.0 的 pH 范围和 40 °C 以下高度稳定，并通过添加亚铁离子 (Fe2+) 进行刺激。在底物特异性方面，BLOX 对亚油酸表现出底物偏好，而亚油酸是大豆中产生氢过氧化物的主要物质。与亚油酸反应，主要产物为13(S)-氢过氧-9,11-十八碳二烯酸；因此，它可以归类为亚油酸酯 13S-LOX 家族 (EC 1.13.11.12)。最终，BLOX的动力学参数（Vmax、Km和kcat）分别为0.124 mM min-1、0.636 mM和12.28 s-1，因此催化效率（kcat/Km）计算为1.93 × 104 M-1·s-1。 BLOX 的这些催化特性有助于了解黑大豆的酶促酸败，并进一步采用生物技术方法来控制和减轻恶化。

更新日期：2020-09-01

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