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Purification and Biochemical Properties of a Glucose-Stimulated ${eta}$-D-Glucosidase Produced by Humicola grisea var. thermoidea Grown on Sugarcane Bagasse
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  • Purification and Biochemical Properties of a Glucose-Stimulated ${eta}$-D-Glucosidase Produced by Humicola grisea var. thermoidea Grown on Sugarcane Bagasse
  • Purification and Biochemical Properties of a Glucose-Stimulated ${eta}$-D-Glucosidase Produced by Humicola grisea var. thermoidea Grown on Sugarcane Bagasse
저자명
Nascimento. Cesar Vanderlei,Souza. Flavio Henrique Moreira,Masui. Douglas Chodi,Leone. Francisco Assis,Peralta. Rosane Marina,Jo
간행물명
The journal of microbiology
권/호정보
2010년|48권 1호|pp.53-62 (10 pages)
발행정보
한국미생물학회
파일정보
정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

The effect of several carbon sources on the production of mycelial-bound ${eta}$-glucosidase by Humicola grisea var. thermoidea in submerged fermentation was investigated. Maximum production occurred when cellulose was present in the culture medium, but higher specific activities were achieved with cellobiose or sugarcane bagasse. Xylose or glucose (1%) in the reaction medium stimulated ${eta}$-glucosidase activity by about 2-fold in crude extracts from mycelia grown in sugarcane bagasse. The enzyme was purified by ammonium sulfate precipitation, followed by Sephadex G-200 and DEAE-cellulose chromatography, showing a single band in PAGE and SDS-PAGE. The ${eta}$-glucosidase had a carbohydrate content of 43% and showed apparent molecular masses of 57 and 60 kDa, as estimated by SDS-PAGE and gel filtration, respectively. The optimal pH and temperature were 6.0 and $50^{circ}C$, respectively. The purified enzyme was thermostable up to 60 min in water at $55^{circ}C$ and showed half-lives of 7 and 14 min when incubated in the absence or presence of 50 mM glucose, respectively, at $60^{circ}C$. The enzyme hydrolyzed p-nitrophenyl-${eta}$-D-glucopyranoside, p-nitrophenyl-${eta}$-D-galactopyranoside, p-nitrophenyl-${eta}$-D-fucopyranoside, p-nitrophenyl-${eta}$-D-xylopyranoside, o-nitrophenyl-${eta}$-D-galactopyranoside, lactose, and cellobiose. The best synthetic and natural substrates were p-nitrophenyl-${eta}$-D-fucopyranoside and cellobiose, respectively. Purified enzyme activity was stimulated up to 2-fold by glucose or xylose at concentrations from 25 to 200 mM. The addition of purified or crude ${eta}$-glucosidase to a reaction medium containing Trichoderma reesei cellulases increased the saccharification of sugarcane bagasse by about 50%. These findings suggest that H. grisea var. thermoidea ${eta}$-glucosidase has a potential for biotechnological applications in the bioconversion of lignocellulosic materials.