Production of enzymes by biotechnological methods: A review

Abstract

Abstract: enzymes are essential biological catalysts widely utilized in diverse industries such as pharmaceuticals, food processing, biofuels, and environmental technology due to their high specificity, efficiency, and eco-friendliness. With increasing global demand, conventional methods of enzyme extraction have become inadequate, leading to the development of advanced biotechnological approaches. The aim of this review is to analyze contemporary strategies for enzyme production using biotechnological methods, evaluate their benefits and limitations, and explore emerging trends aimed at improving enzyme yield, stability, and industrial applicability. The materials and methods used in this review involved a comprehensive analysis of scientific publications over the past three years obtained from international databases, using keywords such as enzyme production, fermentation, recombinant DNA, and protein engineering. The data were synthesized through critical review of original research, review articles, and industrial reports. Biotechnological enzyme production predominantly relies on microbial systems due to their rapid growth, adaptability, and cost-effective fermentation capabilities. Bacteria, fungi, and yeasts are employed in submerged or solid-state fermentation, offering scalability and precise process control. Genetically engineered microorganisms, particularly Escherichia coli, Saccharomyces cerevisiae, and Pichia pastoris, are frequently used as expression hosts, enabling high-yield production and post-translational modifications. The review highlights the significance of recombinant DNA technology, emphasizing cloning strategies, vector design, expression optimization, and fusion proteins for improved purification and secretion. Protein engineering techniques such as rational design and directed evolution allow the fine-tuning of enzyme properties, enhancing thermal stability, pH tolerance, and substrate specificity. These advances have had transformative effects on the pharmaceutical sector, enabling efficient drug synthesis and delivery systems, and supporting sustainable production practices across industries. Although plant- and animal-derived enzymes retain specific industrial roles, microbial enzymes remain dominant due to their robustness and efficiency. Fermentation techniques are central to enzyme production, with submerged fermentation favored for its automation potential, and solid-state fermentation offering higher concentrations and lower costs. The integration of synthetic biology, metagenomics, and AI-driven process control is expected to redefine future enzyme production. In conclusion, biotechnological methods have significantly enhanced the feasibility, scalability, and sustainability of industrial enzyme production. Continued innovation in genetic and protein engineering, coupled with process optimization, promises to expand enzyme applications across emerging industrial sectors.