Revolutionizing recombinant protein production in prokaryotic platforms - Methodologies and advances

Recombinant protein production in prokaryotic systems remains a major topic in biotechnology because of their rapid growth, cost-effectiveness, and ease of genetic manipulation. However, the production of functionally active proteins still faces significant challenges due to folding failures, insolubility, and the lack of the capability of most prokaryotes for complex post-translational processing. This review dwells into both traditional and emerging strategies for optimizing recombinant protein expression in various prokaryotic systems. It also highlights recent advances in genetic engineering and synthetic biology for expanding the toolkit available for protein production, which include refined expression vectors, engineered hosts with improved folding capabilities, and high-throughput screening platforms. Additionally, it provides a thorough discussion of how to optimize heterologous expression using fusion tag approaches, codon bias elimination, promoter and ribosome binding site (RBS) engineering, and chaperone-assisted folding. This review explores diverse prokaryotic expression systems that offer unique advantages for heterologous expression that extend far beyond the limitations of traditional hosts. Additionally, this review also emphasizes the need for the selection of the right expression system and optimizing conditions to fulfill the increasing demands for recombinant protein production in various fields.