Characterization of the monolignol oxidoreductase AtBBE-like protein 15 L182V for biocatalytic applications

Q2 Chemical Engineering

Journal of Molecular Catalysis B-enzymatic Pub Date : 2016-11-01 DOI:10.1016/j.molcatb.2016.10.018

Sabine Pils , Kordula Schnabl , Silvia Wallner , Marko Kljajic , Nina Kupresanin , Rolf Breinbauer , Michael Fuchs , Raquel Rocha , Joerg H. Schrittwieser , Wolfgang Kroutil , Bastian Daniel , Peter Macheroux

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引用次数: 6

Abstract

Monolignol oxidoreductases from the berberine bridge enzyme-like (BBE-like) protein family (pfam 08031) catalyze the oxidation of monolignols to the corresponding aldehydes. In this report, we explore the potential of a monolignol oxidoreductase from Arabidopsis thaliana (AtBBE-like protein 15) as biocatalyst for oxidative reactions. For this study we employed a variant with enhanced reactivity towards oxygen, which was obtained by a single amino acid exchange (L182V). The pH and temperature optima of the purified AtBBE-like protein 15 L182V were determined as well as the tolerance toward organic co-solvents; furthermore the substrate scope was characterized. The enzyme has a temperature optimum of 50 °C and retains more than 50% activity between pH 5 and pH 10 within 5 min. The enzyme shows increased activity in the presence of various co-solvents (10–50% v/v), including acetonitrile, 2-propanol, 1,4-dioxane, and dimethyl sulfoxide. Primary benzylic and primary or secondary allylic alcohols were accepted as substrates. The enantioselectivity E in the oxidation of secondary alcohols was good to excellent (E>34 to >200).

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单脂醇氧化还原酶atbe样蛋白15l182v生物催化应用的表征

来自小檗碱桥酶样(bbe -样)蛋白家族(pfam 08031)的单脂醇氧化还原酶催化单脂醇氧化成相应的醛。在本报告中，我们探讨了拟南芥中单脂醇氧化还原酶(atbbe样蛋白15)作为氧化反应生物催化剂的潜力。在这项研究中，我们采用了一种对氧反应性增强的变体，该变体是通过单氨基酸交换获得的(L182V)。测定纯化的atbe样蛋白15l182v的最适pH和温度，以及对有机共溶剂的耐受性;并对衬底范围进行了表征。该酶的最佳温度为50℃，在pH 5 ~ pH 10范围内5min内保持50%以上的活性。在各种共溶剂(10 - 50% v/v)存在下，酶的活性增加，包括乙腈、2-丙醇、1,4-二氧六环和二甲亚砜。伯苯基醇和伯或仲烯丙醇可作为底物。仲醇氧化对映体选择性E从良好到优异(E>34 ~ E> 200)。

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来源期刊

Journal of Molecular Catalysis B-enzymatic 生物-生化与分子生物学

CiteScore

2.58

自引率

0.00%

发文量

审稿时长

3.4 months

期刊介绍： Journal of Molecular Catalysis B: Enzymatic is an international forum for researchers and product developers in the applications of whole-cell and cell-free enzymes as catalysts in organic synthesis. Emphasis is on mechanistic and synthetic aspects of the biocatalytic transformation. Papers should report novel and significant advances in one or more of the following topics; Applied and fundamental studies of enzymes used for biocatalysis; Industrial applications of enzymatic processes, e.g. in fine chemical synthesis; Chemo-, regio- and enantioselective transformations; Screening for biocatalysts; Integration of biocatalytic and chemical steps in organic syntheses; Novel biocatalysts, e.g. enzymes from extremophiles and catalytic antibodies; Enzyme immobilization and stabilization, particularly in non-conventional media; Bioprocess engineering aspects, e.g. membrane bioreactors; Improvement of catalytic performance of enzymes, e.g. by protein engineering or chemical modification; Structural studies, including computer simulation, relating to substrate specificity and reaction selectivity; Biomimetic studies related to enzymatic transformations.