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Development of the Fundamentals of a Technology for the Production of Tribolium castaneum Recombinant Cathepsin L in Komagataella kurtzmanii Yeastстатья
Информация о цитировании статьи получена из
Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 15 февраля 2024 г.
- Авторы: Serkina A.V., Gubaidullin I.I., Akentiev F.I., Dombrovskii I.A., Filippova I.Yu, Elpidina E.N., Kozlov D.G., Bulushova N.V.
- Журнал: Applied Biochemistry and Microbiology
- Том: 59
- Номер: 8
- Год издания: 2023
- Издательство: Maik Nauka/Interperiodica Publishing
- Местоположение издательства: Russian Federation
- Первая страница: 1078
- Последняя страница: 1088
- DOI: 10.1134/s0003683823080082
- Аннотация: Based on the Komagataella kurtzmanii yeast, a strain producing Tribolium castaneum recombinant procathepsin (rpTcCathL) has been obtained; this strain provided biosynthesis and secretion of at least 0.5 g/L of the target protein as a result of cultivation in flasks on a medium of complex composition. It was determined that the pH of the yeast culture medium (pH ≥ 6.5) is the key factor that influences the biosynthesis and accumulation of procathepsin. A new chromatographic purification scheme was developed, which allows one to obtain recombinant procathepsin with a yield of about 30% and a purity of over 95%. It was shown that complete autocatalytic activation of the proenzyme is achieved within no more than 30 min at a temperature of 37°C and pH 4.0–4.5. The resulting recombinant cathepsin and activated cathepsin at a concentration of 0.5 mg/mL and below could be stored without detectable changes at a temperature of 4°C for at least 2 weeks. Thus, the research made it possible to develop the fundamentals of the technology for obtaining and storing recombinant preparations of the highly purified major digestive cathepsin L from T. castaneum in the form of a proenzyme and mature cathepsin L.
- Добавил в систему: Элпидина Елена Николаевна