Purification and properties of Clostridium thermocellum endoglucanase 5 produced in Escherichia coliстатья
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Дата последнего поиска статьи во внешних источниках: 18 июля 2013 г.
Авторы:
Mosolova T.P.a ,
Kalyuzhnyi S.V.a ,
Varfolomeyev S.D.a ,
Velikodvorskaya G.A.b
Журнал:
Applied Biochemistry and Biotechnology
Том:
42
Номер:
1
Год издания:
1993
Издательство:
Humana Press, Inc.
Местоположение издательства:
United States
Первая страница:
9
Последняя страница:
18
DOI:
10.1007/BF02788898
Аннотация:
Endoglucanase 5 (EG5) has been isolated from the strain of E. coli TGI harboring recombinant plasmid pCU108, which contains the cel5 gene of C. thermocellum. The enzyme has been produced with 98-fold purification and a final yield of 27% by using subsequent twofold high performance ion-exchange chromatography on Mono Q and high performance chromatofocusing on Mono P. The protein has a mol mass of 35 kDa and includes 3 multiple forms with pI 4.4-4.8 as evidenced by analytical gel isoelectrofocusing. EG5 cleaves CMC (Km = 0.097 g/L, Vmax = 8.2 mg/min·mg of protein), amorphous cellulose, xylan, lichenan as a substrate with an optimum temperature of 80‡C and pH 6.0 and Avicel (Km = 18.2 g/L, Vmax = 0.035 mg/min·mg of protein) with an optimum temperature of 60‡C and pH 6.O. Cellobiose in concentrations up to 200 Μg/mL do not inhibit the hydrolysis of CMC by EG5, but 10-30 Μg/mL of glucose significantly decrease the activity of this enzyme. The stimulating role of calcium chloride and concentration of protein in the system has been demonstrated for Avicel hydrolysis by EG5. © 1993 Humana Press Inc.
Добавил в систему:
Калюжный Сергей Владимирович