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利用包埋的 l-阿拉伯糖异构酶在酵母孢子中生产 l-核酮糖。

Production of l-Ribulose Using an Encapsulated l-Arabinose Isomerase in Yeast Spores.

机构信息

Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology , Jiangnan University , Wuxi , Jiangsu 214122 , People's Republic of China.

School of Food Science and Technology , Jiangnan University , Wuxi , Jiangsu 214122 , People's Republic of China.

出版信息

J Agric Food Chem. 2019 May 1;67(17):4868-4875. doi: 10.1021/acs.jafc.9b00640. Epub 2019 Apr 16.

Abstract

The rare sugar l-ribulose is produced from the abundant sugar l-arabinose by enzymatic conversion. An l-arabinose isomerase (AI) from Geobacillus thermodenitrificans was efficiently expressed and encapsulated in Saccharomyces cerevisiae spores. Deletion of the yeast OSW2 gene, which causes a mild defect in the integrity of the spore wall, substantially improved the activity of encapsulated AI, without damaging its superior enzymatic properties of thermostability, pH tolerance,and resistance toward SDS and proteinase treatments. In a 10 mL reaction, 100 mg of dry AI encapsulated in spores produced 250 mg of l-ribulose from 1 g of l-arabinose, indicating a 25% conversion rate. Notably, the product of l-ribulose was directly purified from the reaction solution with an approximately 91% recovery using a Ca ion exchange column. Our results describe not only a facile approach for the production of l-ribulose but also a useful strategy for the enzymatic conversion of rare sugars in "Izumoring".

摘要

稀有糖 l-核酮糖可由丰富的糖 l-阿拉伯糖经酶转化得到。一种来自嗜热脱硫杆菌的 l-阿拉伯糖异构酶(AI)在酿酒酵母孢子中得到高效表达和包封。酵母 OSW2 基因的缺失导致孢子壁完整性的轻微缺陷,极大地提高了包封 AI 的活性,而不会损害其卓越的热稳定性、pH 耐受性以及对 SDS 和蛋白酶处理的抗性等酶学特性。在 10 mL 反应中,100 mg 干燥的 AI 包封在孢子中,可将 1 g 的 l-阿拉伯糖转化为 250 mg 的 l-核酮糖,转化率为 25%。值得注意的是,产物 l-核酮糖可直接使用 Ca 离子交换柱从反应溶液中直接纯化,回收率约为 91%。我们的结果不仅描述了一种生产 l-核酮糖的简便方法,还为“ Izumoring ”中稀有糖的酶转化提供了一种有用的策略。

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