vestichИзвестия Национальной академии наук Беларуси. Серия химических наукProceedings of the National Academy of Sciences of Belarus, Chemical Series1561-83312524-2342The Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1561-8331-2025-61-4-327-337vestich-985Research ArticleБИООРГАНИЧЕСКАЯ ХИМИЯBIOORGANIC CHEMISTRYГармонизация кодонов как способ увеличения уровня экспрессии функционально-активного CYP17A1 человекаCodon harmonization as a way to increase the expression level of functionally active human CYP17A1ШаладоноваМ. И.ShaladonovaM. I.

Шаладонова Марина Игоревна – аспирант, младший научный сотрудник

ул. Академика Купревича, 5/2, 220084, Минск

Shaladonova Marina I. – Postgraduate Student, Junior Researcher

5/2, Academician Kuprevich Str., 220084, Minsk

m.shaladonova@gmail.comДиченкоЯ. В.DzichenkaYa. V.

Диченко Ярослав Владимирович – кандидат химических наук, ведущий научный сотрудник

ул. Академика Купревича, 5/2, 220084, Минск

Dzichenka Yaraslau V. – Ph. D. (Chemistry), Leading Researcher

5/2, Academician Kuprevich Str., 220084, Minsk

dichenko@iboch.byЩурВ. В.ShchurV. V.

Щур Вероника Владимировна – кандидат химических наук, старший научный сотрудник

ул. Академика Купревича, 5/2, 220084, Минск

Shchur Veronika V. – Ph. D. (Chemistry), Senior Researcher

5/2, Academician Kuprevich Str., 220084, Minsk

shchur931@gmail.comТравкинаМ.TrawkinaM.

Травкина Мария – научный сотрудник

ул. Академика Купревича, 5/2, 220084, Минск

Trawkina Maria – Researcher

5/2, Academician Kuprevich Str., 220084, Minsk

maria_trawkina@iboch.byУсановС. А.UsanovS. A.

Усанов Сергей Александрович – член-корреспондент, доктор химических наук, профессор

ул. Академика Купревича, 5/2, 220084, Минск

Usanov Sergei A. – Corresponding Member, Dr. Sc. (Chemistry), Professor

5/2, Academician Kuprevich Str., 220084, Minsk

usanov@iboch.byИнститут биоорганической химии Национальной академии наук БеларусиInstitute of Bioorganic Che- mistry of the National Academy of Sciences of Belarus202503122025614327337Copyright © Шаладонова М.И., Диченко Я.В., Щур В.В., Травкина М., Усанов С.А., 20252025Шаладонова М.И., Диченко Я.В., Щур В.В., Травкина М., Усанов С.А.Shaladonova M.I., Dzichenka Y.V., Shchur V.V., Trawkina M., Usanov S.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestichem.belnauka.by/jour/article/view/985

CYP17A1 является ключевым ферментом в биосинтезе глюкокортикоидов и андрогенов, а дисфункция данного фермента сопровождается тяжелыми нарушениями в организме, в том числе гормон-зависимыми злокачественными новообразованиями (рак предстательной железы, рак молочной железы). Впервые проведена гармонизация нуклеотидной последовательности гена СYP17A1 человека и осуществлена оптимизация методики его выделения и очистки из бактериальных клеток различных штаммов. Установлено, что гармонизация кодонов гена, кодирующего СYP17A1 человека, позволяет увеличить уровень экспрессии целевого белка на 28 %. Проведенный анализ гармонизации кодонов позволил выявить, что редкими кодонами представлены только определенные аминокислоты (A, C, D, G, I, V, Y), также редкие кодоны встречаются в аминокислотных остатках активного центра белка. В ходе исследования доказано, что гармонизированный белок взаимодействует с природным субстратом СYP17A1 – прогестероном аналогично оптимизированному белку и обладает функциональной активностью. Полученные результаты дают основание считать, что гармонизация кодонов для СYP17A1 человека является методом, способным оптимизировать получение препаративных количеств терапевтически значимого фермента c сохранением его каталитической активности.

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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The authors declare that there are no conflicts of interest present.