Microbial amidases: Characterization, advances and biotechnological applications

Rajendra Singh , Refana Shahul , Vijay Kumar , Ashok Kumar Yadav , Praveen Kumar Mehta
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Abstract

The amidases (EC 3.5.1.4) are versatile hydrolase biocatalysts that have been the attention of academia and industries for stereo-selective synthesis and bioremediation. These are categorized based on the amino acid sequence and substrate specificity. Notably, the Signature amidase family is distinguished by a characteristic signature sequence, GGSS(S/G)GS, which encompasses highly conserved Ser-Ser-Lys catalytic residues, and the amidases belonging to this family typically demonstrate a broad substrate spectrum activity. The amidases classified within the nitrilase superfamily possess distinct Glu-Lys-Cys catalytic residues and exhibit activity towards small aliphatic substrates. Recent discoveries have underscored the potential role of amidases in the degradation of toxic amides present in polymers, insecticides, and food products. This expands the horizons for amidase-mediated biodegradation of amide-laden pollutants and fosters sustainable development alongside organic synthesis. The burgeoning global production facilities are expected to drive a heightened demand for this enzyme, attributable to its promising chemo-, regio-, and enantioselective hydrolysis capabilities for a variety of amides. Advances in protein engineering have enhanced the catalytic efficiency, structural stability, and substrate selectivity of amidases. Concurrently, the heterologous expression of amidase genes sourced from thermophiles has facilitated the development of highly stable amidases with significant industrial relevance. Beyond their biotransformation capabilities concerning amides, through amido-hydrolase and acyltransferase activities, recent investigations have illuminated the potential of amidase-mediated degradation of amide-containing pollutants in soil and aquatic environments. This review offers a comprehensive overview of recent advancements pertaining to microbial amidases (EC 3.5.1.4), focusing on aspects such as their distribution, gene mining methodologies, enzyme stability, protein engineering, reusability, and biocatalytic efficacy in organic synthesis and biodegradation.
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微生物酰胺酶:特性、进展和生物技术应用。
酰胺酶(EC 3.5.1.4)是一种多功能的水解酶生物催化剂,在立体选择合成和生物修复方面受到学术界和工业界的关注。这些是根据氨基酸序列和底物特异性分类的。值得注意的是,Signature amidase家族的特征序列GGSS(S/G)GS包含高度保守的Ser-Ser-Lys催化残基,属于该家族的酰胺酶通常具有广泛的底物光谱活性。腈酶超家族中的酰胺酶具有独特的Glu-Lys-Cys催化残基,并对小脂肪底物具有活性。最近的发现强调了酰胺酶在降解聚合物、杀虫剂和食品中存在的有毒酰胺方面的潜在作用。这扩大了酰胺酶介导的含酰胺污染物生物降解的范围,并促进了有机合成的可持续发展。由于该酶具有对多种酰胺的化学、区域和对映体选择性水解能力,全球生产设施的迅速发展预计将推动对该酶的更高需求。蛋白质工程技术的进步提高了酰胺酶的催化效率、结构稳定性和底物选择性。同时,来自嗜热菌的氨基酶基因的异源表达促进了高度稳定的氨基酶的发展,具有重要的工业意义。除了它们对酰胺的生物转化能力之外,通过酰胺水解酶和酰基转移酶的活性,最近的研究已经阐明了酰胺酶介导的土壤和水生环境中含酰胺污染物降解的潜力。本文综述了微生物酰胺酶(EC 3.5.1.4)的分布、基因挖掘方法、酶的稳定性、蛋白质工程、可重复利用以及在有机合成和生物降解中的生物催化作用等方面的研究进展。
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