Industrially produced pullulanases with thermostability: Discovery, engineering, and heterologous expression.
Industrially produced pullulanases with thermostability: Discovery, engineering, and heterologous expression.
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DOI：
10.1016/j.biortech.2019.01.098
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发表时间：
2019-04
影响因子：
11.4
通讯作者：
Xinye Wang;Y. Nie;Yan Xu
中科院分区：
文献类型：
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作者：
Xinye Wang;Y. Nie;Yan Xu
普鲁兰酶（EC 3.2.1.41）是一种淀粉脱支酶，广泛用于水解淀粉、支链淀粉、支链淀粉和其他低聚糖中的α-1，6-糖苷键，在食品、酿造和制药工业中具有应用潜力。虽然进行了广泛的研究来发现和表达支链淀粉酶，但只有少数具有工业应用所需的特性。这对挖掘新的酶源，基于序列/结构设计蛋白质以及调节表达提出了挑战。我们在这里审查鉴定极端和嗜温微生物作为工业普鲁兰酶的来源，具有理想的特性，包括耐酸性，热稳定性和嗜冷性。本文介绍了普鲁兰酶定点突变和序列/结构指导蛋白质工程的最新进展。此外，我们还讨论了普鲁兰酶在原核和真核微生物系统中的异源表达，并讨论了表达元件的有效性及其对酶生产的调节。最后，我们指出未来的研究需要开发所需的工业支链淀粉酶。
Pullulanases (EC 3.2.1.41) are well-known starch-debranching enzymes widely used to hydrolyze α-1,6-glucosidic linkages in starch, pullulan, amylopectin, and other oligosaccharides, with application potentials in food, brewing, and pharmaceutical industries. Although extensive studies are done to discover and express pullulanases, only few are available with desirable characteristics for industrial applications. This raises the challenge to mine new enzyme sources, engineer proteins based on sequence/structure, and regulate expressions. We review here the identification of extremophilic and mesophilic microbes as sources of industrial pullulanases with desirable characteristics, including acid-resistance, thermostability, and psychrotrophism. We present current advances in site-directed mutagenesis and sequence/structure-guided protein engineering of pullulanases. In addition, we discuss heterologous expression of pullulanases in prokaryotic and eukaryotic microbial systems, and address the effectiveness of the expression elements and their regulation of enzyme production. Finally, we indicate future research needs to develop desired industrial pullulanases.