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Optimization of RNA Structure Leads to Increased Biosynthesis of E. coli L-asparaginase when its Gene is Overexpressedстатья
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: 23 января 2026 г.
- Авторы: Shayfutdinov R.R., Orlova N.A., Vorobiev I.I.
- Журнал: Applied Biochemistry and Microbiology
- Том: 61
- Номер: 6
- Год издания: 2025
- Издательство: Maik Nauka/Interperiodica Publishing
- Местоположение издательства: Russian Federation
- Первая страница: 1063
- Последняя страница: 1074
- DOI: 10.1134/s0003683825602148
- Аннотация: A highly efficient expression system for recombinant Escherichia coli type II L-asparaginase (EC 3.5.1.1) has been developed based on a synthetic gene optimized for the energy of secondary structure hairpins formation in the 5'-region of mRNA. A producer strain, E. coli BL21[DE3]/pET28a-AsnSYN was created, providing an accumulation of up to 291 ± 9 mg/L of enzymatically active protein (44.5 ± 2.6 mg/(L AU)) when cultivated in stirred flasks, which is 50% higher than the control strain with a natural gene at an induction time of 3 hours. Optimization of the codon composition of the gene made it possible to increase the energy of secondary mRNA structure formation in the 5'-region from –70 to –47 kcal/mol, which, as we assume, contributed to improved translation efficiency. For the developed producer strain, asparaginase that meets pharmacopoeial purity requirements can be obtained with a total yield of ≥25% and a specific activity of >250 IU/mg. The preparation contains no visible impurities according to electrophoresis data and less than 3% multimeric forms according to size exclusion chromatography data. The results obtained demonstrate the promise of using synthetic genes with optimized DNA structure for the industrial production of therapeutic enzymes.
- Добавил в систему: Орлова Надежда Александровна