Department of Microbiology and Bioprocess Technology, Institute of Biochemistry, University of Leipzig, Leipzig, Germany.
Department of Microbiology and Bioprocess Technology, Institute of Biochemistry, University of Leipzig, Leipzig, Germany.
Adv Appl Microbiol. 2014;89:267-305. doi: 10.1016/B978-0-12-800259-9.00007-X.
Thermophilic actinomycetes produce enzymes capable of hydrolyzing synthetic polyesters such as polyethylene terephthalate (PET). In addition to carboxylesterases, which have hydrolytic activity predominantly against PET oligomers, esterases related to cutinases also hydrolyze synthetic polymers. The production of these enzymes by actinomycetes as well as their recombinant expression in heterologous hosts is described and their catalytic activity against polyester substrates is compared. Assays to analyze the enzymatic hydrolysis of synthetic polyesters are evaluated, and a kinetic model describing the enzymatic heterogeneous hydrolysis process is discussed. Structure-function and structure-stability relationships of actinomycete polyester hydrolases are compared based on molecular dynamics simulations and recently solved protein structures. In addition, recent progress in enhancing their activity and thermal stability by random or site-directed mutagenesis is presented.
嗜热放线菌产生的酶能够水解合成聚酯,如聚对苯二甲酸乙二醇酯(PET)。除了羧基酯酶外,角质酶相关的酯酶也能水解合成聚合物,它们主要对 PET 低聚物具有水解活性。本文描述了放线菌产生这些酶及其在异源宿主中的重组表达,并比较了它们对聚酯底物的催化活性。评估了分析合成聚酯酶解的酶分析方法,并讨论了描述酶异质水解过程的动力学模型。基于分子动力学模拟和最近解决的蛋白质结构,比较了放线菌聚酯水解酶的结构-功能和结构-稳定性关系。此外,还介绍了通过随机或定点突变来提高其活性和热稳定性的最新进展。