Regulation of catalytic activity of recombinant L-asparaginase from rhodospirillum rubrum by conjugation with a PEG-chitosan copolymer

Malakhova, M.A.; Pokrovskaya, M.V.; Alexandrova, S.S.; Sokolov, N.N.; Kudryashova, E.V.

Авторы: Malakhova M.A., Pokrovskaya M.V., Alexandrova S.S., Sokolov N.N., Kudryashova E.V.
Журнал: Moscow University Chemistry Bulletin
Том: 59
Номер: 4
Год издания: 2018
Издательство: Allerton Press Inc.
Местоположение издательства: United States
Первая страница: 297
Последняя страница: 304
DOI: 10.3103/S0027131418040065
Аннотация: A new approach for the regulation of catalytic properties of the medically significant enzyme L-asparaginase is suggested based on the formation of conjugates with PEG-chitosan (chitoPEGylation). The efficiency of this approach is demonstrated using recombinant L-asparaginase from Rhodospirillum rubrum (RrA). This preparation is immunologically different from the one used in medical practice preparations of L-asparaginase from E. coli, which offers a promising alternative for applications in the case of hypersentsitivity development. The low level of activity of RrA towards L-glutamine, which decreases significantly the chance of side effects developing, is an advantage of RrA. The technique for the synthesis of the RrA conjugates with PEG-chitosan (chitoPEGylation) of a varying modification degree is developed. It is established that conjugation of RrA with PEG-chitosan increased the specific activity of the enzyme in comparison with the native one. The activity changes from 56 IU/mg (for the native enzyme) to 61–72 IU/mg (for the conjugates) depending on the degree of chitosan PEGylation. The secondary structure of the Rhodospirillum rubrum asparaginase conjugates with PEG-chitosan is examined using CD- and IR-spectroscopy. It is found that the enzyme structure changed only slightly as a result of conjugation with PEG-chitosan: the content of α-helices changed from 36% (for the native enzyme) to 30–33% (for the conjugates). The content of β-structures changed from 15% (for the native enzyme) to 18% (for the conjugate). The obtained data open new opportunities for the synthesis of L-asparaginase preparations with improved biocatalytic properties.
Добавил в систему: Кудряшова Елена Вадимовна

	ИСТИНА	Войти в систему Регистрация
	ИСТИНА ИНХС РАН
	Главная Поиск Статистика О проекте Помощь

ИСТИНА

ИСТИНА ИНХС РАН

Regulation of catalytic activity of recombinant L-asparaginase from rhodospirillum rubrum by conjugation with a PEG-chitosan copolymerстатья

Прикрепленные файлы