Volume 11,Issue 3
Advances in Polymerase Engineering and Their Biotechnological Applications
DNA and RNA polymerases are essential enzymes responsible for nucleic acid replication, transcription, and amplification, and they play indispensable roles in molecular biology, biotechnology, and synthetic biology. However, the natural properties of polymerases are often limited by their evolutionary origins, resulting in constraints in catalytic efficiency, substrate compatibility, thermostability, and replication fidelity. In recent years, advances in protein engineering and structural biology have enabled the development of diverse strategies to enhance polymerase performance. This review summarizes recent progress in polymerase engineering aimed at structural optimization and functional enhancement. Major engineering strategies, including directed evolution, rational and semi-rational design, computational protein engineering, and domain fusion approaches, are discussed in terms of their principles and applications. In addition, the rapidly expanding applications of engineered polymerases are highlighted, particularly in molecular diagnostics, enzymatic DNA synthesis, artificial genetic systems, and medical and industrial biotechnology. Overall, the continuous integration of high-throughput screening technologies, structural analysis, and artificial intelligence–assisted design is expected to accelerate the development of next-generation polymerases with improved performance and expanded catalytic capabilities. These advances will further promote innovations in synthetic biology, nucleic acid diagnostics, genome engineering, and the development of novel biotechnological tools.
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