Characterization of inulolytic enzymes from the Jerusalem artichoke-derived Glutamicibacter mishrai NJAU-1

文献类型： 外文期刊

第一作者： Lian, Dan

作者： Lian, Dan;Zhuang, Shuo;Shui, Chen;Zheng, Shicheng;Liang, Mingxiang;Ma, Yanhong;Sun, Zongjiu;Porras-Dominguez, Jaime R.;Van den Ende, Wim;Porras-Dominguez, Jaime R.;Van den Ende, Wim;Oner, Ebru Toksoy

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关键词： Bio fructose; Exo-inulinase; Glutamicibacter mishrai; Fructan; Jerusalem artichoke

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：5.56； 五年影响因子：5.365 ）

ISSN： 0175-7598

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收录情况： SCI

摘要： The rhizosphere context of inulin-accumulating plants, such as Jerusalem artichoke (Helianthus tuberosus), is an ideal starting basis for the discovery of inulolytic enzymes with potential for bio fructose production. We isolated a Glutamicibacter mishrai NJAU-1 strain from this context, showing exo-inulinase activity, releasing fructose from fructans. The growth conditions (pH 9.0; 15 degrees C) were adjusted, and the production of inulinase by Glutamicibacter mishrai NJAU-1 increased by 90% (0.32 U/mL). Intriguingly, both levan and inulin, but not fructose and sucrose, induced the production of exo-inulinase activity. Two exo-inulinase genes (inu/ and inu2) were cloned and heterologously expressed in Pichia pastoris. While INU2 preferentially hydrolyzed longer inulins, the smallest fructan 1-kestose appeared as the preferred substrate for INU1, also efficiently degrading nystose and sucrose. Active site docking studies with GFn- and Fn-type small inulins (G is glucose, F is fructose, and n is the number of beta (2-1) bound fructose moieties) revealed subtle substrate differences between INU1 and INU2. A possible explanation about substrate specificity and INU's protein structure is then suggested.

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