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Title: Properties of cellulase immobilized on agarose gel with spacer

Abstract

Cellulase produced by fungus Trichoderma viride was immobilized on agarose beads (Sepharose 4B) activated by cyanogen bromide and also on activated agarose beads that contained spacer arm (activated Ch-Sepharose 4B and Affi-Gel 15). The CMCase activity retained by immobilized cellulase on activated Sepharose containing the spacer tended to be higher than that immobilized without spacer, although the extent of protein immobilization was lower. Also, the higher substrate specificity for cellulase immobilized on beads with spacer was obtained for cellobiose, acid-swollen cellulose, or cellulose powder. The hydrolysis product from their substrates was mainly glucose. 10 references.

Authors:
; ; ; ;
Publication Date:
Research Org.:
National Food Research Institute, Ibaraki, Japan
OSTI Identifier:
6561637
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biotechnol. Bioeng.; (United States); Journal Volume: 28:12
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; 59 BASIC BIOLOGICAL SCIENCES; CELLULASE; ENZYME ACTIVITY; IMMOBILIZED ENZYMES; AGAR; CELLOBIOSE; CHEMICAL ACTIVATION; ENZYMATIC HYDROLYSIS; GELS; GLUCOSE; PH VALUE; POWDERS; PRODUCTION; PROTEINS; SUBSTRATES; TRICHODERMA VIRIDE; ALDEHYDES; CARBOHYDRATES; CHEMICAL REACTIONS; COLLOIDS; DECOMPOSITION; DISACCHARIDES; DISPERSIONS; ENZYMES; FUNGI; GLYCOSYL HYDROLASES; HEXOSES; HYDROLASES; HYDROLYSIS; LYSIS; MONOSACCHARIDES; O-GLYCOSYL HYDROLASES; OLIGOSACCHARIDES; ORGANIC COMPOUNDS; PLANTS; POLYSACCHARIDES; SACCHARIDES; SOLVOLYSIS; TRICHODERMA; 090222* - Alcohol Fuels- Preparation from Wastes or Biomass- (1976-1989); 550700 - Microbiology; 140504 - Solar Energy Conversion- Biomass Production & Conversion- (-1989)

Citation Formats

Chim-anage, P., Kashiwagi, Y., Magae, Y., Ohta, T., and Sasaki, T. Properties of cellulase immobilized on agarose gel with spacer. United States: N. p., 1986. Web. doi:10.1002/bit.260281215.
Chim-anage, P., Kashiwagi, Y., Magae, Y., Ohta, T., & Sasaki, T. Properties of cellulase immobilized on agarose gel with spacer. United States. doi:10.1002/bit.260281215.
Chim-anage, P., Kashiwagi, Y., Magae, Y., Ohta, T., and Sasaki, T. 1986. "Properties of cellulase immobilized on agarose gel with spacer". United States. doi:10.1002/bit.260281215.
@article{osti_6561637,
title = {Properties of cellulase immobilized on agarose gel with spacer},
author = {Chim-anage, P. and Kashiwagi, Y. and Magae, Y. and Ohta, T. and Sasaki, T.},
abstractNote = {Cellulase produced by fungus Trichoderma viride was immobilized on agarose beads (Sepharose 4B) activated by cyanogen bromide and also on activated agarose beads that contained spacer arm (activated Ch-Sepharose 4B and Affi-Gel 15). The CMCase activity retained by immobilized cellulase on activated Sepharose containing the spacer tended to be higher than that immobilized without spacer, although the extent of protein immobilization was lower. Also, the higher substrate specificity for cellulase immobilized on beads with spacer was obtained for cellobiose, acid-swollen cellulose, or cellulose powder. The hydrolysis product from their substrates was mainly glucose. 10 references.},
doi = {10.1002/bit.260281215},
journal = {Biotechnol. Bioeng.; (United States)},
number = ,
volume = 28:12,
place = {United States},
year = 1986,
month =
}
  • Aspergillus niger KKS, isolated from a farmland near Suwon, was immobilized on Celite and polyurethane foams. Enzyme activities produced by the immobilized cell system in a bubble column were higher than that of shake-flask culture. The enzyme productivities were twice as high. {Beta}-Glucosidase, {Beta}-xylosidase, and xylanase activities obtained in a bubble column were significant when the ground rice straw was used as a substrate. 9 refs., 2 figs., 3 tabs.
  • The thermodynamics of interaction of (R)- and (S)-propranolol between an acetic acid buffer (pH = 4.7 and 5.5) and the protein cellobiohydrolase I immobilized on silica gel was studied between 5 and 45{degree}C. The equilibrium data were fitted to a biLangmuir adsorption isotherm with excellent agreement. One of the two Langmuir contributions is the same for both enantiomers and accounts for the nonspecific interactions between these compounds and most sites on the surfaces (type-I, nonselective sites). It has a large saturation capacity. The second contribution accounts for the chiral selective interactions (type-II sites). It has a lower monolayer capacity thanmore » the first. The interaction enthalpy and entropy on type-I sites are -1.1 kcal/mol and +0.1 cal/(mol K), respectively. For type-II sites, they are -1.9 kcal/mol and -2.6 cal/(mol K), respectively, for (R)-propranolol and +1.6 kcal/mol and +11.6 cal/(mol K), respectively, for (S)-propranolol at pH = 5.5. This explains why at this pH the retention time of the less-retained R enantiomer decreases with increasing temperature, while the retention time of the S enantiomer increases, causing a large increase of the separation factor when the temperature is raised from 5 to 45{degree}C. The saturation capacity of the chiral contributions depends strongly on the pH, and the retention times of both enantiomers decrease with increasing temperature at pH = 4.7. 46 refs., 6 figs., 5 tabs.« less
  • Ethanol yields were 2.1 (P = 0.06) to 2.3 (P = 0.01) times higher in simultaneous saccharification and fermentation (SSF) reactions of microcrystalline cellulose when cellulase was physisorbed on silica nanoparticles compared to enzyme in solution. In SSF reactions, cellulose is hydrolyzed to glucose by cellulase while yeast simultaneously ferments glucose to ethanol. The 35 C temperature and the presence of ethanol in SSF reactions are not optimal conditions for cellulase. Immobilization onto solid supports can stabilize the enzyme and promote activity at non-optimum reaction conditions. Mock SSF reactions that did not contain yeast were used to measure saccharification productsmore » and identify the mechanism for the improved ethanol yield using immobilized cellulase. Cellulase adsorbed to 40 nm silica nanoparticles produced 1.6 times (P = 0.01) more glucose than cellulase in solution in 96 h at pH 4.8 and 35 C. There was no significant accumulation (<250 {mu}g) of soluble cellooligomers in either the solution or immobilized enzyme reactions. This suggests that the mechanism for the immobilized enzyme's improved glucose yield compared to solution enzyme is the increased conversion of insoluble cellulose hydrolysis products to soluble cellooligomers at 35 C and in the presence of ethanol. The results show that silica-immobilized cellulase can be used to produce increased ethanol yields in the conversion of lignocellulosic materials by SSF.« less
  • Continuous cellulase production by Trichoderma viride QM 9123, immobilized in 6 mm diameter, spherical, stainless steel biomass support particles, has been achieved using a medium containing glucose as the main carbon source. Experiments were carried out in a 10-L spouted bed fermentor. In this type of reactor recycled broth is used to create a jet at the base of a bed of particles, causing the particles to spout and circulate. During the circulation, particles pass through a region of high shear near the jet inlet. This effectively prevents a buildup of excess biomass and thus enables steady-state conditions to bemore » achieved during continuous operation. Continuous production of cellulase was achieved at significantly higher yield and productivity than in conventional systems. At a dilution rate of 0.15 h/sup -1/ (nominal washout rate for freely suspended cells is 0.012 h/sup -1/), the yield of cellulase on glucose was 31% higher than that measured during batch operation, while the volumetric productivity (31.5 FPA U/L x h) was 53% greater than in the batch system. The specific cellulase productivity of the immobilized cells was more than 3 times that of freely suspended cells, showing that diffusional limitations can be beneficial. This offers significant opportunity for the further development of biomass support particles and associated bioreactors.« less
  • Cellulase (Cellulosin AC-8) was immobilized on poly-L-glutamic acid. This immobilized cellulase (IC) is water soluble in the neutral and alkaline solutions, where IC has the activity, while IC can be made insoluble by lowering the pH so that it can be recovered from the reaction mixture with its activity. The optimum pH and temperature were determined to be 5.5 and 55 degrees C, respectively. The stability of IC against change in the pH and temperature was improved by the immobilization. Solvolysis of 3N-NaOH-treated cellulose, with IC under the optimum conditions found here, led to the production of low-molecular-weight compounds. 15more » references.« less