vuzbiochemiИзвестия вузов. Прикладная химия и биотехнологияProceedings of Universities. Applied Chemistry and Biotechnology2227-29252500-1558ИРНИТУ10.21285/achb.1015JBQXVQvuzbiochemi-1600Research ArticleФИЗИКО-ХИМИЧЕСКАЯ БИОЛОГИЯPHYSICOCHEMICAL BIOLOGYВыявление целлюлозолитической активности штаммов энтомопатогенной бактерии Bacillus thuringiensisCellulolytic activity of strains of the entomopathogenic bacterium Bacillus thuringiensishttps://orcid.org/0000-0001-5401-0579КрыжкоА. В.KryzhkoA. V.

Крыжко Анастасия Владимировна, к.с-х.н., ведущий научный сотрудник

295453, г. Симферополь, ул. Киевская, 150

Anastasiia V. Kryzhko, Cand. Sci. (Agriculture), Leading Researcher

150, Kievskaya St., Simferopol, 295453

nk_lib@mail.ruНаучно-исследовательский институт сельского хозяйства КрымаРоссияResearch Institute of Agriculture of CrimeaRussian Federation2026020420261617281Copyright © Крыжко А.В., 20262026Крыжко А.В.Kryzhko A.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vuzbiochemi.elpub.ru/jour/article/view/1600

Исследование механизмов разложения целлюлозы у штаммов Bacillus thuringiensis представляет интерес для поиска новых перспективных продуцентов целлюлаз. Способность Cry3Аа белков Bacillus thuringiensis связываться с целлюлазой способствует увеличению стабильности токсинов в кишечнике насекомых. Определение целлюлазной активности проводили качественно при помощи красителя конго красного и количественным методом А.П. Синицына. Скорость разрушения целлюлозы измеряли методом Н.Л. Кристенсена. Ген целлюлазной активности Bacillus thuringiensis идентифицировали c помощью специфичных праймеров. Целью данной работы являлось изучение целлюлозолитической активности и детектирование генов целлюлазы в штаммах Bacillus thuringiensis Крымской коллекции микроорганизмов. В ходе исследования установлено, что все исследованные штаммы Bacillus thuringiensis с коэффициентом подобия, равным 92%, образовывали монофилетическую группу, характеризующуюся высокой степенью гомологии по гену целлюлазы и принадлежащую преимущественно к серотипам thuringiensis, kurstaki и dendrolimus. Максимальная целлюлазная активность, достигающая в среднем 0,25 мкг/мл, характерна для штаммов Bacillus thuringiensis, принадлежащих к серотипу thuringiensis, а максимальной целлюлозолитической активностью (в среднем до 80%) обладали штаммы, принадлежащие к серотипам kurstaki и thuringiensis. Подтверждены выраженные целлюлозолитические свойства для штаммов Bacillus thuringiensis 685, 792, 800, 926, 942, 989, 072, 0105, 0162, 0307, 0308, 0326, 0371, 0411, 0441, 0452, 0532, 0541, 41Н1, 109Н10,12-91, 177, 992. Обработка чернозема южного культурой штаммов Bacillus thuringiensis 072, 0105, 0326 способствует увеличению целлюлозолитических свойств чернозема южного до 99,5% относительно контроля.

Studying the mechanisms of cellulose breakdown in Bacillus thuringiensis strains is of interest in the search for potential cellulase producers. Also, the ability of Cry3Aa proteins in Bacillus thuringiensis to bind to cellulase increases the stability of toxins in the insect gut. In the study, cellulase activity was determined qualitatively using Congo red dye and quantitatively via a method developed by A.P. Sinitsyn. The rate of cellulose breakdown was measured employing the Christensen method. The cellulase gene in Bacillus thuringiensis was identified using specific primers. The study aimed to examine cellulolytic activity and detect cellulase genes in Bacillus thuringiensis strains from the Crimean collection of microorganisms. At a similarity coefficient of 92%, all the examined Bacillus thuringiensis strains were found to form a monophyletic group characterized by a high cellulase gene homology, which comprises predominantly the serotypes thuringiensis, kurstaki, and dendrolimus. Maximum cellulase activity, reaching an average of 0.25 μg/mL, was observed in Bacillus thuringiensis strains belonging to the thuringiensis serotype, while the maximum cellulolytic activity (on average up to 80%) was exhibited by strains belonging to the kurstaki and thuringiensis serotypes. Pronounced cellulolytic properties were confirmed in the Bacillus thuringiensis strains 685, 792, 800, 926, 942, 989, 072, 0105, 0162, 0307, 0308, 0326, 0371, 0411, 0441, 0452, 0532, 0541, 41Н1, 109Н10, 12-91, 177, and 992. Treatment of southern black earth with the Bacillus thuringiensis strains 072, 0105, and 0326 was found to increase its cellulolytic properties up to 99.5% relative to the control.

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The authors declare that there are no conflicts of interest present.