RU2011123739A - Протеазы, содержащие одну или несколько комбинируемых мутаций - Google Patents

Протеазы, содержащие одну или несколько комбинируемых мутаций Download PDF

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RU2011123739A
RU2011123739A RU2011123739/10A RU2011123739A RU2011123739A RU 2011123739 A RU2011123739 A RU 2011123739A RU 2011123739/10 A RU2011123739/10 A RU 2011123739/10A RU 2011123739 A RU2011123739 A RU 2011123739A RU 2011123739 A RU2011123739 A RU 2011123739A
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subtilisin
positions
variant
detergent
seq
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Луис Г. КАСКАО-ПЕРЕЙРА
Дэвид А. ЭСТЕЛЛ
МЛ. Джеймс Т. КЕЛЛИС
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Abstract

1. Выделенный вариант субтилизина, который является зрелой формой, обладающей протеолитической активностью и включающей замену в двух или более положениях, выбираемых из положений 24, 45, 101, 109, 118, 213 и 217, нумерация которых соответствует аминокислотной последовательности субтилизина BPN' B. amyloliquefaciens, представленной как SEQ ID NO:1.2. Выделенный вариант субтилизина по п.1, который имеет относительный индекс продуктивности уровня экспрессии белка (ТСА PI) и/или индекс продуктивности активности по удалению пятен (BMI PI), больший или равный 0,5.3. Выделенный вариант субтилизина по п.1 или 2, где указанный субтилизинпредставляет собой GG36 и где указанная замена в двух или более положениях выбрана из S24Q, S24E, S24L, S24R, R45Q, R45E, R45L, S101Q, S101E, S101L, S101R, Q109E, Q109L, Q109R, G118Q, G118E, G118L, G118R, T213Q, T213L, T213R, T213E, L217Q и L217E, соответствующих положениям субтилизина BPN' SEQ ID NO:1.4. Выделенный вариант субтилизина по любому из п.1 или 2, где указанный субтилизинпредставляет собой GG36 и где указанная замена в двух или более положениях представляет собой S24Q-R45Q-S101Q-G118Q-T213Q, соответствующую положениям субтилизина BPN' SEQ ID NO:1.5. Выделенная нуклеиновая кислота, кодирующая вариант субтилизина по любому из пп.1-4.6. Экспрессирующий вектор, содержащий нуклеиновую кислоту по п.5.7. Клетка-хозяин, содержащая экспрессирующий вектор по п.6.8. Очищающая композиция, содержащая по меньшей мере один вариант субтилизина по любому из пп.1-4.9. Очищающая композиция по п.8, которая является детергентом.10. Очищающая композиция по п.9, в которой указанный детергент является сверхсильным жидким или сухим стиральным детергентом.11. Очищающая композиция по п.9, в которой указанный детергент является детергентом для мыть�

Claims (15)

1. Выделенный вариант субтилизина Bacillus, который является зрелой формой, обладающей протеолитической активностью и включающей замену в двух или более положениях, выбираемых из положений 24, 45, 101, 109, 118, 213 и 217, нумерация которых соответствует аминокислотной последовательности субтилизина BPN' B. amyloliquefaciens, представленной как SEQ ID NO:1.
2. Выделенный вариант субтилизина по п.1, который имеет относительный индекс продуктивности уровня экспрессии белка (ТСА PI) и/или индекс продуктивности активности по удалению пятен (BMI PI), больший или равный 0,5.
3. Выделенный вариант субтилизина по п.1 или 2, где указанный субтилизин Bacillus представляет собой GG36 и где указанная замена в двух или более положениях выбрана из S24Q, S24E, S24L, S24R, R45Q, R45E, R45L, S101Q, S101E, S101L, S101R, Q109E, Q109L, Q109R, G118Q, G118E, G118L, G118R, T213Q, T213L, T213R, T213E, L217Q и L217E, соответствующих положениям субтилизина BPN' SEQ ID NO:1.
4. Выделенный вариант субтилизина по любому из п.1 или 2, где указанный субтилизин Bacillus представляет собой GG36 и где указанная замена в двух или более положениях представляет собой S24Q-R45Q-S101Q-G118Q-T213Q, соответствующую положениям субтилизина BPN' SEQ ID NO:1.
5. Выделенная нуклеиновая кислота, кодирующая вариант субтилизина по любому из пп.1-4.
6. Экспрессирующий вектор, содержащий нуклеиновую кислоту по п.5.
7. Клетка-хозяин, содержащая экспрессирующий вектор по п.6.
8. Очищающая композиция, содержащая по меньшей мере один вариант субтилизина по любому из пп.1-4.
9. Очищающая композиция по п.8, которая является детергентом.
10. Очищающая композиция по п.9, в которой указанный детергент является сверхсильным жидким или сухим стиральным детергентом.
11. Очищающая композиция по п.9, в которой указанный детергент является детергентом для мытья посуды.
12. Очищающая композиция по любому из пп.8-11, дополнительно содержащая один или несколько дополнительных ферментов или производных ферментов, выбираемых из гемицеллюлаз, целлюлаз, пероксидаз, протеаз, металлопротеаз, ксиланаз, липаз, фосфолипаз, эстераз, пергидролаз, кутиназ, пектиназ, пектат-лиаз, маннаназ, кератиназ, редуктаз, оксидаз, фенолоксидаз, липоксигеназ, лигниназ, пуллуланаз, танназ, пентозаназ, маланаз, β-глюканаз, арабинозидаз, гиалуронидаз, хондроитиназ, лакказ и амилаз или их смесей.
13. Очищающая композиция по любому из пп.8-11, дополнительно содержащая по меньшей мере один стабилизатор.
14. Очищающая композиция, содержащая по меньшей мере 0,0001 мас.% по меньшей мере одного варианта субтилизина по любому из пп.1-4 и необязательно по меньшей мере один приемлемый дополнительный ингредиент.
15. Способ очистки, который включает стадии:
а) контактирования поверхности и/или изделия, содержащего ткань, с очищающей композицией по любому из пп.7-14; и
b) необязательно промывки и/или прополаскивания указанной поверхности или изделия.
RU2011123739/10A 2008-11-11 2009-11-10 Протеазы, содержащие одну или несколько комбинируемых мутаций RU2560978C2 (ru)

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US11354508P 2008-11-11 2008-11-11
US61/113,545 2008-11-11
US21880209P 2009-06-19 2009-06-19
US61/218,802 2009-06-19
PCT/US2009/063837 WO2010056653A2 (en) 2008-11-11 2009-11-10 Proteases comprising one or more combinable mutations

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US9096838B2 (en) 2015-08-04
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CN103361196A (zh) 2013-10-23
RU2560978C2 (ru) 2015-08-20
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CA2743302A1 (en) 2010-05-20
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US20140308736A1 (en) 2014-10-16
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WO2010056653A2 (en) 2010-05-20
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