RECOMBINANT PLASMID pHpFDH_HisA, PROVIDING SYNTHESIS IN ESCHERICHIA coli CELLS OF FORMATE DEHYDROGENASE OF METHYLOTROPHIC YEAST HANSENULA POLYMORPHA (HpFDH_HisA) AND RECOMBINANT STRAIN ESCHERICHIA coli BL21(DE3)/pHpFDH_HisA VKPM B-14559 - HpFDH_HisA PRODUCER Russian patent published in 2024 - IPC C12N15/53 C12N15/11 C12N1/21 C12N15/70 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, particularly to genetic engineering. Was constructed recombinant plasmid DNA pHpFDH_HisA, containing modified gene hpfdh_hisa, coding mutant `NAD⁺-dependent formate dehydrogenase (EC 1.2.1.2, HpFDH_HisA) with additional sequence of six histidine residues and one residue of alanine on N-terminus of enzyme. Addition of an additional sequence of His₆Ala to the N-terminus of the enzyme increases thermal stability and enables to obtain preparations of a highly purified enzyme in a single step. To ensure high efficiency of expression in the target producer strain in the modified HpFDH_HisA gene, a TAATAA triplet tandem is used as a stop codon instead of the TGA triplet in the initial gene. Obtained plasmid enables expression of the included gene hpfdh_hisa in Escherichia coli cells.

EFFECT: as a result of transformation of the Escherichia coli BL21(DE3) strain by the proposed plasmid, a new recombinant producer strain is obtained Escherichia coli BL21(DE3))/pHpFDH_HisA VKPM B-14559, providing overexpression of HpFDH_HisA in active form.

2 cl, 6 dwg, 4 tbl, 4 ex