Продукция субтилизиновых протеаз в бактериях и дрожжах

В настоящей работе мы рассматриваем прогресс в изучении и модификации субтилизиновых протеаз. Несмотря на длительное время применения микробиальных протеаз и значительное число работ, посвященных их исследованию, поиск новых генов протеаз, создание продуцентов и развитие методов их применения остаются актуальными, о чем говорит высокий уровень цитирования публикаций, описывающих протеазы и их продуценты. На данный класс ферментов приходится максимальный объем производства промышленных белков в мире, что объясняет большой интерес к нему. Это говорит о чрезвычайно высокой важности получения собственных технологий их производства. В статье представлены сведения о классификации субтилизинов, истории их открытия и дальнейших работ по оптимизации их свойств. Дан обзор классов субтилизиновых протеаз и родственных им ферментов. Проанализированы проблемы поиска и отбора субтилаз из природных штаммов различных микроорганизмов, пути и особенности их модификации и используемые при этом методы генетической инженерии. Детально изучены методы оптимизации продукции промышленных субтилаз у различных штаммов, касающихся важнейших аспектов культивирования: состава среды, времени культивирования, влияния температуры и pH. Приводятся результаты последних исследований по техникам культивирования – глубинному и твердофазному культивированию. На основании рассмотренных литературных данных можно заключить, что в настоящее время практически не применяются нативные, т. е. обнаруженные в природе ферменты, в связи с решающими преимуществами, предоставляемыми белками, модифицированными при помощи генной инженерии и обладающими улучшенными свойствами: термостабильностью, общей устойчивостью к детергентам и специфической – к различным окислителям, высокой активностью в разных диапазонах температур, независимостью от ионов, стабильностью в отсутствие кальция и т. д. Большинство субтилизиновых протеаз синтезируется в штаммах-продуцентах, относящихся к разным видам рода Bacillus. В то же время ведутся работы по адаптации синтеза этих ферментов в других микроорганизмах, в частности дрожжей Pichia pastoris.

субтилизин, субтилаза, протеаза, щелочная сериновая протеаза, Pichia pastoris, Bacillus subtilis, биотехнология, генетическая инженерия, культивирование

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