Codon harmonization as a way to increase the expression level of functionally active human CYP17A1

CYP17A1 is a key enzyme in the biosynthesis of glucocorticoids and androgens, and its dysfunction is associated with severe disorders, including hormone-dependent malignant neoplasms (prostate cancer and breast cancer). In this study, the codon harmonization of the human CYP17A1 gene was performed for the first time, and the method for its expression and purification from bacterial cells of various strains was optimized. It was found that harmonization of codons of the gene encoding human CYP17A1 increases the expression level of the target protein by 28 %. The analysis of codon harmonization revealed that only certain amino acids (A, C, D, G, I, V, Y) are represented by rare codons, and rare codons are also found in the amino acid residues of the active center of the protein. The study demonstrated that the harmonized protein interacts with the natural substrate of CYP17A1, progesterone, similarly to the optimized protein and exhibits functional activity. The results obtained indicate that codon harmonization for human CYP17A1 is an effective approach for producing preparative quantities of the therapeutically significant enzyme while maintaining its catalytic activity.

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