Sequencing analysis of intracellular inserted formate dehydrogenase gene and uninserted recombinant komagataella phaffii
In this study, the intracellular metabolic pathway of K. phaffii was modified. Firstly, the formate dehydrogenase gene of the dissimilar pathway was knocked out, and then the formate dehydrogenase of Candida boidinii (CbFDH)was inserted to construct an autotrophic pathway of CO2 fixation in the cell of recombinant K. phaffii. CO2 was reduced to formate by NADH electrochemical regeneration system, and the formate was further used to induce the expression of foreign proteins by recombinant K. phaffii. Finally, the expression of recombinant protein was induced by CO2 substrate, and transcriptomic analysis was performed on the recombinant K. phaffii. RNA-seq technology was an efficient approach to investigate the transcriptome, which was verified as a reliable method by several researchers. The analysis of the metabolic pathway of recombinant protein expression induced by formate synthesis on CO2 substrate will contribute to the early realization of the goal of carbon neutrality, and has positive significance for the application of K. phaffii in the safe production of foreign proteins.