Expression of Recombinant Taq DNA Polymerase in Oligotrophic Klebsiella oxytoca: A Novel Approach to Industrial Enzyme Production

Abstract

The demand for high-efficiency DNA polymerases in molecular biology and diagnostic applications has led to the exploration of novel microbial hosts for enzyme production. This study investigates the expression of recombinant Taq DNA polymerase in Klebsiella oxytoca, an oligotrophic bacterium known for its minimal nutrient requirements and robust growth in diverse environments. By leveraging the metabolic versatility and adaptive capabilities of K. oxytoca, we aimed to establish a cost-effective and sustainable method for producing Taq polymerase at an industrial scale. The recombinant K. oxytoca was engineered using a plasmid vector containing the Taq polymerase gene under the control of a strong promoter. Optimal expression conditions were identified, including the appropriate induction time and temperature, leading to high yields of active enzyme. The Taq DNA polymerase was successfully expressed in a standard LB medium and at a concentration of 0.1% (v/v). Expressed Taq DNA polymerases were characterized by SDS-PAGE and PCR activity analyses were performed. The same processes were also carried out in scale-up studies, and it was investigated whether TaqDNA polymerase production in Klebsiella oxytoca was suitable for industry. This approach not only reduces production costs but also aligns with green chemistry principles by utilizing a host organism that thrives on minimal resources. Our findings suggest that oligotrophic K. oxytoca is a promising candidate for the biotechnological production of recombinant enzymes, offering an innovative pathway for enhancing industrial enzyme manufacturing processes.