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Designing a Thermostable Mini-Intein for Intein-Mediated Purification of Recombinant Proteins and Peptidesстатья
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: 6 августа 2025 г.
- Авторы: Karanov Andrey A., Zayats Evgeniy A., Kostromina Maria A., Abramchik Yulia A., Sharafutdinova Aleksandra R., Surkova Maria S., Zamyatnin Andrey A., Esipov Roman S.
- Журнал: Biochemistry (Moscow)
- Том: 90
- Номер: 6
- Год издания: 2025
- Издательство: Pleiades Publishing, Ltd
- Местоположение издательства: Road Town, United Kingdom
- Первая страница: 818
- Последняя страница: 827
- DOI: 10.1134/s0006297925600358
- Аннотация: This paper reports the design of a thermostable temperature-activated mini-intein based on the full-length intein DnaE1 from Thermus thermophilus HB27 (TthDnaE1). We performed rational design of three mini-inteins TthDnaE1 Δ272, Δ280, and Δ287 through deletion mutations in the full-length intein sequence. Two mini-inteins (Δ272 and Δ280) were capable of efficient protein splicing at temperatures above 50°C. The most active mini-intein with the Δ280 deletion was selected as a platform for further design of a self-cleaving carrier of affinity tags through single-point mutagenesis. Three mutations - C1A, D405G, and the combined C1A/D405G - were introduced to inhibit N-terminal extein cleavage and extein ligation. As a result, the mini-intein Δ280 with double mutation C1A/D405G displayed the highest efficiency of C-terminal extein cleavage with temperature optimum around 60°C. Thus, we constructed thermostable temperature-activated mini-intein capable of efficient protein splicing or cleavage of the C-terminal extein. The engineered TthDnaE1 Δ280 C1A/D405G mini-intein can serve as a basis for the development of new expression system for intein-mediated production of pharmaceutically relevant recombinant proteins and peptides.
- Добавил в систему: Замятнин Андрей Александрович
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