METHOD OF OBTAINING BIOLOGICALLY ACTIVE RECOMBINANT PROTEINS Russian patent published in 2023 - IPC C07K14/00 C07K19/00 C12N15/63 

FIELD: biotechnology.

SUBSTANCE: method of obtaining biologically active recombinant proteins containing up to 4 disulfide bonds. The method involves the development of a genetic construct encoding a precursor of the target protein, containing in its composition, arranged in the order of enumeration from the N-terminus to the C-terminus, the sequence of the self-associating peptide L₆KD, including 6 leucine residues following one after another, the sequence of SUMO protein, the product of Smt3 gene of the yeast Saccharomyces cerevisiae, and the sequence of the target protein, the first N-terminal residue of which, following the sequence of the SUMO protein, is different from proline. The created genetic construct is used to obtain the target protein by transforming a suitable recipient strain with this construct, cultivating the transformed strain under conditions that ensure the expression of the genetic construct and microbiological synthesis of the precursor of the target target protein in the form of insoluble inclusion bodies, followed by isolation of the synthesized inclusion bodies from cells of the producer strain, releasing the soluble target protein from their composition using a suitable SUMO-specific proteinase derivative, separating the residues of insoluble inclusion bodies by centrifugation, and further purifying the released target protein using molecular biology techniques, without the use of renaturation.

EFFECT: invention provides effective production of biologically active recombinant target proteins produced using microbiological synthesis without using any denaturation/renaturation procedures.

1 cl, 5 dwg, 2 tbl, 16 ex