Paryzhak S. Сloning genes of formaldehyde dehydrogenase FLD1 and formaldehyde reductase ADH1 of the yeast Hansenula polymorpha and gene-engineering construction of producers of dehydrogenase and reductase of formaldehyde.

Perspective gene-engineered yeast clones, over-producing formaldehyde dehydrogenase (FdDH), originated from the recipient thermotolerant methylotrophic yeast H. polymorpha with multi-copy integration of the FLD1 gene and with elevated resistance to formaldehyde (up to 20 mM) have been developed. To obtain recombinant strains of methylotrophic yeast H. polymorpha capable to over-produce formaldehyde reductase, ORF of ADH1 gene from chromosomal DNA of S. cerevisiae was cloned under the control of strong constitutive promotеr of GAP1 H. рolymorpha gene. ADH1 gene of H. рolymorpha, coding one of the ADH isoforms, which reveals formaldehyde-reductase activity was identified and cloned. Intact cells of the gene-engineered thermotolerant methylotrophic yeast H. polymorpha, with a high content of NAD+- and glutathione-dependent FdDH, were used as the biorecognition elements for amperometric assay of formaldehyde.