FIELD: biotechnology.
SUBSTANCE: invention relates to recombinant plasmid DNA pEst877 determining expression of the polypeptide with the activity of esterase P. cryohalolentis K5T on the cell surface with the molar weight of 3.64 Md (5.519 kb) consisting of NcoI/XhoI - DNA fragment of plasmid pET20b(+) with length of 3.654 kb, comprising a promoter T7lac, the bacteriophage T7 transcription terminator, the gene of bla β-lactamase which determines the stability of the cells transformed with the plasmid pEst877 to ampicillin, the site ori of replication initiation, signal sequence pelB of pectate lyase B Erwinia carotovora; and NcoI/XhoI-fragment of DNA with the size of 1.865 kb containing fusion gene Est877 encoding the amino acid sequences of esterase P. cryohalolentis K5T with an additional amino acid residue of aspartic acid after the site of cleavage of signal peptide and auto-transporter P. cryohalolentis K5T in a single reading frame. The invention also relates to the bacterial strain E. coli BL21(DE3)pLysS/pEst877 - producer of polypeptide with the activity of esterase P. cryohalolentis K5T on the cell surface.
EFFECT: invention enables to obtain highly active esterase with wide substrate specificity.
2 cl, 3 dwg, 7 ex
