Characterization of formate dehydrogenase from Trametes versicor for formate production from CO2 gas
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Enzymatic CO<sub>2 </sub>reduction has been reported as a promising approach to greenhouse gas fixation. There are few biocatalysts capable of biological CO<sub>2</sub> fixation, e.g. pyruvate decarboxylase, carbonic anhydrase, and formate dehydrogenase. Formate dehydrogenase(FDH) can reduce CO<sub>2</sub> to formate without organic chemicals, and formate can be sequentially reduced to formaldehyde and methanol by coupling aldehyde dehydrogenase and alcohol dehydrogenase reactions. Therefore, FDH has been widely adopted inCO<sub>2 </sub>reducing reaction.<br /> In this study, we researched CO<sub>2</sub>-reducing activities of FDH derived from wood rot fungi, <i>Trametes versicolor</i>. Microbial species, <i>T. versicolor</i>, were separated in National institute of Forest Science. The fungi were grown in a potato dextrose agar medium. For the transformation, the host, <i>Shizosaccaromyces pombe </i>of wild type, was obtained from Bioneer(Korea). Total RNA was extracted with Hybrid-R (GeneAll) from <i>T. versicolor </i>mycelium. cDNAs were synthezied by AccuPower (Bioneer) as recommended by the manufacturer. For sub-cloning, PCR of fragments (1.2 kb) were amplified with pairs of TvFDH ORF primers(Foword: 5’-ATGCTCGCCGGCATCT-3’, Reverse: 5’-TCACTTGCGCTGGCCA-3’). The amplified fragments were sub-cloned into a pCR2.1-TOPO (Invitrogen, Carlsbad, CA, USA) vector and the plasmid DNA was sequenced. The amplified fragments were digested by Not 1 and Bam 1 and then inserted into a pSLF272 vector (Bioneer, Korea). The recombinant plasmid was amplified in the Escherichia coli DH5α, and then extracted by mini prep kit. The recombinant plasmids were chemically (lithium acetate method) transformed into yeast<i> S. pombe</i>. Ligation products, transformants, were named as pSLF272-TvFDH. Selection of transformants were performed in the minimal medium, EMM. Expression and purification of the TvFDHs is being performed to obtain pure enzymes.
Keywords: Climate change, Research, Genetic resources, Agriculture, Innovation
ID: 3604933