Ecological geneticsEcological geneticsЭкологическая генетика1811-09322411-9202Eco-Vector245910.17816/ecogen11258-72ArticlesСтатьиResearch ArticleBiosynthesis of hexaand pentameric chitooligosaccharides using n-acetylglucoseaminyl transferase from rhizobial bacteriaБиосинтез гекса- и пентамерных хитоолигосахаридов с помощью n-ацетилглюкозаминилтрансферазы ризобиальных бактерийLeppyanenIrina ViktorovnaЛеппяненИрина Викторовна

scientist

младший научный сотрудник, лаборатория молекулярной и клеточной биологии.

leppyanen_irina@rambler.ruArtamonovaTatyana OlegovnaАртамоноваТатьяна Олеговна

scientist

научный сотрудник

artamonova@nanobio.spbstu.ruLopatinSergey AleksandrovichЛопатинСергей Александрович

PhD

к. х. н., старший научный сотрудник

lopatin@biengi.ac.ruVarlamovValeriy PetrovichВарламовВалерий Петрович

doctor of science

д. х. н., заведующий лабораторией, профессор

varlamov@biengi.ac.ruTikhonovichIgor AnatolyevichТихоновичИгорь Анатольевич

doctor of science, professor, academician of RAAS

д. б. н., профессор, академик РАСХН

contact@arriam.spb.ruDolgikhYelena AnatolyevnaДолгихЕлена Анатольевна

PhD, group leader

к. б. н., ведущий научный сотрудник, лаборатория молекулярной и клеточной биологии

dol2helen@yahoo.comAll-Russia Research Institute for Agricultural MicrobiologyГНУ ВНИИСХМSt.Petersburg State Polytechnical UniversityСанкт-Петербургский государственный политехнический университетCenter “Bioengineering” of RASУчреждение Российской академии наук Центр «Биоинженерия» РАН15062013112VOL 11, NO2 (2013)ТОМ 11, №2 (2013)587230032016Copyright ©; 2013, Leppyanen I.V., Artamonova T.O., Lopatin S.A., Varlamov V.P., Tikhonovich I.A., Dolgikh Y.A.Copyright ©; 2013, Леппянен И.В., Артамонова Т.О., Лопатин С.А., Варламов В.П., Тихонович И.А., Долгих Е.А.2013Leppyanen I.V., Artamonova T.O., Lopatin S.A., Varlamov V.P., Tikhonovich I.A., Dolgikh Y.A.Леппянен И.В., Артамонова Т.О., Лопатин С.А., Варламов В.П., Тихонович И.А., Долгих Е.А.http://creativecommons.org/licenses/by/4.0https://journals.eco-vector.com/ecolgenet/article/view/2459

Chitooligosaccharides find wide application that determines considerable interest in their use. Enzymatic synthesis of hexa-N-acetylchitohexaose and penta-N-acetylchitopentaose using N-acetyl-glucoseaminyl transferase enzyme possessing unique features from rhizobial bacteria Rhizobium sp. GRH2 and M. loti has been performed in E. coli cells. Cultivation of bacteria E. coli expressing the appropriate recombinant enzyme resulted in synthesis of significant amounts of desired chitooligosaccharides (milligrams per liter). Analysis of synthesized chitooligosacchairdes by methods of high performance liquid chromatography and mass-spectrometry confirmed the conformity of the synthesized compounds to standards.

Хитоолигосахариды находят широкое практическое применение, что определяет интерес к изучению возможности биосинтеза этих соединений. В данной работе осуществлен синтез гекса-N-ацетилхитогексаозы и пента-N-ацетилхитопентаозы в клетках E. coli с использованием уникального фермента N-ацетилглюкозаминилтрансферазы клубеньковых бактерий Rhizobium sp. GRH2 и M. loti. При культивировании бактерий E. coli, экспрессирующих рекомбинантные ферменты, в присутствии субстрата N-ацетилглюкозамина, уровень синтеза хитоолигосахаридов достигал нескольких миллиграмм на литр жидкой культуры. Анализ полученных соединений с помощью методов ВЭЖХ и масс-спектрометрии подтвердил их соответствие стандартам.

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