Adsorption and synergism of cellobiohydrolase I and II of Trichoderma reesei during hydrolysis of microcrystalline cellulose

Journal Article · Sat Nov 05 04:00:00 EST 1994 · Biotechnology and Bioengineering; (United States)

DOI:https://doi.org/10.1002/bit.260440907· OSTI ID:6778819

Medve, J; Tjerneld, F ^[1]; Staahlberg, J ^[2]

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1. Univ. of Lund (Sweden). Dept. of Biochemistry
2. Univ. of Uppsala (Sweden). Dept. of Molecular Biology

Hydrolysis of microcrystalline cellulose (Avicel) by cellobiohydrolase I and II (CBH I and II) from Trichoderma reesei has been studied. Adsorption and synergism of the enzymes were investigated. Experiments were performed at different temperatures and enzyme/substrate ratios using CBH I and CBH II alone and in reconstituted equimolar mixtures. Fast protein liquid chromatography (FPLC) analysis was found to be an accurate and reproducible method to follow the enzyme adsorption. A linear correlation was found between the conversion and the amount of adsorbed enzyme when Avicel was hydrolyzed by increasing amounts of CBH I and/or CBH II. CBH I had lower specific activity compared to CBH II although, over a wide concentration range, more CBH I was adsorbed than CBH II. Synergism between the cellobiohydrolases during hydrolysis of the amorphous fraction of Avicel showed a maximum as a function of total enzyme concentration. Synergism measured as a function of bound enzyme showed a continuous increase, which indicates that by decreasing the distance between the two enzymes the synergism is enhanced. The adsorption process for both enzymes was slow. Depending on the enzyme/substrate ratio it took 30--90 min to reach 95% of the equilibrium binding. The amount of bound enzyme decreased with increasing temperature. The two enzymes compete for the adsorption sites but also bind to specific sites. Stronger competition for adsorption sites was shown by CBH I.

• OSTI ID: 6778819
• Journal Information: Biotechnology and Bioengineering; (United States), Journal Name: Biotechnology and Bioengineering; (United States) Vol. 44:9; ISSN BIBIAU; ISSN 0006-3592
• Country of Publication: United States
• Language: English

Related Subjects

09 BIOMASS FUELS
090900* -- Biomass Fuels-- Processing-- (1990-)
ADSORPTION
CARBOHYDRATES
CELLULOSE
CHEMICAL REACTIONS
CHROMATOGRAPHY
DECOMPOSITION
ENZYMATIC HYDROLYSIS
ENZYME ACTIVITY
ENZYMES
EUMYCOTA
FUNGI
HYDROLASES
HYDROLYSIS
LIQUID COLUMN CHROMATOGRAPHY
LYSIS
ORGANIC COMPOUNDS
PLANTS
POLYSACCHARIDES
PROTEINS
SACCHARIDES
SEPARATION PROCESSES
SOLVOLYSIS
SORPTION
SYNERGISM
TRICHODERMA
TRICHODERMA VIRIDE