Srivastava Nitin, Khare Sunil Kumar
Enzyme and Microbial Biochemistry Laboratory, Department of Chemistry, Indian Institute of Technology Delhi, Hauz Khas, New Delhi, 110016, India.
Department of Biological Sciences, Indian Institute of Science Education and Research Kolkata, Mohanpur, 741246, West Bengal, India.
Appl Biochem Biotechnol. 2025 May 15. doi: 10.1007/s12010-025-05270-9.
Microbial alkaline proteases are versatile enzymes chiefly employed in various industrial sectors, viz., food processing, detergents, leather, textile, pharmaceutical industries. However, the existing bottlenecks, such as lower enzyme yields, stability, purification, specificity, and catalytic rates, bring resistance toward their industrial suitability. The robust microbes are prominent sources of stable enzymes. However, further challenges may exist, such as low yield, difficult purification, and lesser enzymatic efficiency. With the advent of advanced genomic and enzyme engineering approaches, such bottlenecks can be overcome. Initially, the microbial genomes can be used as novel repositories for stable enzyme sequences for further heterologous production with higher enzymatic yields and an easier purification process. Moreover, enzyme improvement through directed evolution and rational engineering could enhance enzyme stability and efficiency. Currently, conventional enzyme improvement methods are increasingly replaced by Artificial Intelligence-Machine Learning (AI-ML) and computational data-driven tools that provide precise information for tailoring enzymes for industrial endeavors. Hence, the current review encompasses a deliberate study of microbial alkaline proteases, their major industrial applications, and the bottlenecks in their commercial implementations. Further, it presents in-detailed solutions, including genetic and enzyme engineering, and insights toward incorporating advanced tools like AI-ML and de novo enzyme engineering to subside the existing challenges.
微生物碱性蛋白酶是用途广泛的酶，主要应用于各个工业领域，即食品加工、洗涤剂、皮革、纺织、制药行业。然而，现有的瓶颈，如酶产量较低、稳定性差、纯化困难、特异性不强以及催化速率低等，使其在工业适用性方面受到限制。健壮的微生物是稳定酶的重要来源。然而，可能还存在其他挑战，如产量低、纯化困难以及酶效率较低等。随着先进的基因组学和酶工程方法的出现，这些瓶颈可以被克服。首先，微生物基因组可作为稳定酶序列的新库，用于进一步的异源生产，以获得更高的酶产量和更简便的纯化过程。此外，通过定向进化和理性工程进行酶的改进可以提高酶的稳定性和效率。目前，传统的酶改进方法正越来越多地被人工智能-机器学习（AI-ML）和计算数据驱动的工具所取代，这些工具为工业应用定制酶提供了精确信息。因此，本综述对微生物碱性蛋白酶、其主要工业应用以及商业应用中的瓶颈进行了深入研究。此外，还详细介绍了包括基因工程和酶工程在内的解决方案，以及关于引入AI-ML和从头酶工程等先进工具以克服现有挑战的见解。
