Rapid and simplified procedure for purification of a cellulase from Fusarium lini

Journal Article · Sun Jan 01 04:00:00 EST 1984 · Biotechnol. Bioeng.; (United States)

DOI:https://doi.org/10.1002/bit.260260109· OSTI ID:7034377

Vaidya, M; Seeta, R; Mishra, C; Deshpande, V; Rao, M

An endo-..beta..-1,4-glucanase (EC3.2.1.4) was obtained in high yields in purified form from a culture filtrate of Fusarium lini by an extremely simple method. The method consists of precipitation of the culture filtrate with ammonium sulphate (290 g/L), followed by chromatography of the precipitated fraction on Biogel P-150. The purification is based on the unusual property of the enzyme being eluted after cytochrome C, even though its molecular weight is 2.8 x 10/sup 4/ (by SDS PAGE). The yield of pure enzyme was 6.8 mg/L culture broth. The homogeneity of the enzyme was established by ultracentrifugation, isoelectric focusing, and electrophoresis in polyacrylamide gels containing SDS. The enzyme was isoelectric at pH 8.3 and contained 2.9% carbohydrate. The K/sub M/ value for carboxymethyl (CM) cellulose was 11.6 mg/mL. The enzyme showed high viscosity reducing activity towards CM cellulose but very low activity with Walseth cellulose and crystalline celluloses such as Avicel and cotton. The purified enzyme has activity towards xylan. The amino acid analysis showed a predominance of acidic and neutral amino acids and low contents of histidine, arginine, and methionine. One-half of the cysteine content was 11 residues/mol enzyme, and no free-SH group was detectable.

• Research Organization: National Chemical Lab., Poona, India
• OSTI ID: 7034377
• Journal Information: Biotechnol. Bioeng.; (United States), Journal Name: Biotechnol. Bioeng.; (United States) Vol. 26:1; ISSN BIBIA
• Country of Publication: United States
• Language: English

Related Subjects

09 BIOMASS FUELS
140504* -- Solar Energy Conversion-- Biomass Production & Conversion-- (-1989)
AMINO ACIDS
CARBOHYDRATES
CARBOXYLIC ACIDS
CELLULASE
CELLULOLYTIC ACTIVITY
CELLULOSE
CENTRIFUGATION
CHEMICAL PROPERTIES
CHEMICAL REACTIONS
CHROMATOGRAPHY
DECOMPOSITION
ELECTROPHORESIS
ENZYMATIC HYDROLYSIS
ENZYME ACTIVITY
ENZYMES
FUNGI
FUSARIUM
GLYCOSYL HYDROLASES
HYDROLASES
HYDROLYSIS
LYSIS
O-GLYCOSYL HYDROLASES
ORGANIC ACIDS
ORGANIC COMPOUNDS
PARASITES
PLANTS
POLYSACCHARIDES
PURIFICATION
SACCHARIDES
SEPARATION PROCESSES
SOLVOLYSIS
ULTRACENTRIFUGATION