Adsorption and desorption of cellulase components during enzymatic hydrolysis of steamed shirakamba (Betula platyphylla Skatchev) wood

doi:10.1016/0922-338X(91)90316-9

Journal of Fermentation and Bioengineering

Volume 72, Issue 2, 1991, Pages 96-100

Journal of Fermentat...

https://doi.org/10.1016/0922-338X(91)90316-9 Get rights and content

Abstract

The adsorption behavior of Trichoderma viride cellulases on steamed shirakamba (Betula platyphylla Skatchev) wood was studied. Substrate/enzyme (S/E) ratio, hydrolysis time and saccharification extent as well as the presence or absence of lignin in the substrate were taken into consideration as factors affecting the enzyme adsorption. Filter paperase (FPase) components were adsorbed more selectively than other cellulase components on the substrate. The presence of lignin in the steaamed hardwood tended to slow down the enzyme adsorption, but it did not appear to restrict the extent of hydrolysis of the carbohydrate moiety. The changes in the composition of the enzyme preparation during the course of hydrolysis were analyze by fast protein liquid chromatography (FPLC). The irreversible adsorption of specific cellulase components was not observed in the prolonged hydrolysis of steamed shirakamba wood containing abundant lignin. The removal of lignin from steamed hardwood prior to hydrolysis is not always necessary for the recycling of cellulolytic enzymes.

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Cited by (15)

Valorizing recycled paper sludge by a bioethanol production process with cellulase recycling
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Citation Excerpt :
In addition to that, the fraction of enzymes remaining adsorbed to the final solid were around 14%, which is clearly inferior comparatively to the other enzymes. This suggests a lower solid-adsorption efficiency for these enzymes (Gomes et al., 2015; Ishihara et al., 1991; Lindedam et al., 2013), which is explained by the lack of a cellulose-binding module (CBM) in this class of enzymes. Previous results suggested some heterogeneity in what concerns the stability and adsorption behavior of different cellulolytic components over the entire recycling experiment.
The feasibility of cellulase recycling in the scope of bioethanol production from recycled paper sludge (RPS), an inexpensive byproduct with around 39% of carbohydrates, is analyzed. RPS was easily converted and fermented by enzymes and cells, respectively. Final enzyme partition between solid and liquid phases was investigated, the solid-bound enzymes being efficiently recovered by alkaline washing. RPS hydrolysis and fermentation was conducted over four rounds, recycling the cellulases present in both fractions. A great overall enzyme stability was observed: 71, 64 and 100% of the initial Cel7A, Cel7B and β-glucosidase activities, respectively, were recovered. Even with only 30% of fresh enzymes added on the subsequent rounds, solid conversions of 92, 83 and 71% were achieved for the round 2, 3 and 4, respectively. This strategy enabled an enzyme saving around 53–60%, while can equally contribute to a 40% reduction in RPS disposal costs.
Key features of pretreated lignocelluloses biomass solids and their impact on hydrolysis
2010, Bioalcohol Production: Biochemical Conversion of Lignocellulosic Biomass
Prior to biological conversion of lignocellulosic biomass to ethanol or other products, natural barriers developed to protect plants must be overcome to realize efficient enzymatic hydrolysis, and a few pretreatment technologies are effective in inexpensively accomplishing this task through heating with chemicals. Over the years, changes in a number of structural and compositional attributes of biomass have been postulated to explain how pretreatment enhances enzymatic hydrolysis performance, but the complexity of biomass has always confounded development of a unified theory that can unequivocally predict how pretreated biomass solids will respond to enzymes. However, sugar release can be viewed as ultimately governed by two factors: 1) access of enzymes to cellulose and hemicellulose and 2) the effectiveness of enzymes attached to the surface in breaking down these carbohydrate chains to sugars and/or their oligomers. In this review, this perspective of enzyme access and effectiveness is applied to findings reported in the literature to provide a framework for understanding how various features in pretreated biomass solids could affect deconstruction of cellulose and hemicellulose to sugars and their yields.
Integrated process for total utilization of wood components by steam-explosion pretreatment
1998, Biomass and Bioenergy
Various species of hardwood chips were subjected to steam-explosion at 180–230°C for 1–20 min. On steaming, hemicellulose was hydrolyzed partially becoming extractable with water, and lignin was degraded by extensive cleavage of α- and β-aryl ether linkages becoming extractable with organic solvents and/or dilute alkali. The three main components, hemicellulose, lignin, and cellulose, of steam-exploded woods were fractionated by successive extraction with water and 90% dioxane. The water extracts were decolored and purified by chromatography on synthetic adsorbents and ion exchange resins, yielding a mixture of xylose and xylooligosaccharides (DP2∼10). The xylooligosaccharides were hydrolyzed to xylose with hemicellulolytic enzymes immobilized on ceramics having controlled pore size. The yield of xylose was 10–20% based on starting materials. The extracted amounts of lignin were different among wood species. Syringyl lignin became more soluble than guaiacyl lignin on steaming. The lignin extracted was converted to thermoplastic materials, lignin-pitch, by phenolysis followed by heat treatment under vacuum. The lignin-pitch was well spun into fine filaments at a speed of 500–1000 m min⁻¹ in the temperature range 150–190°C using the melt-spinning method. The filaments were carbonized on heating from room temperature up to 1000°C in a stream of nitrogen. The carbon fiber was obtained in a yield of more than 40% based on the starting materials. The physical properties of the lignin- based carbon fiber was equivalent to a commercial carbon fiber made from petroleum pitch. The residual fibers, mainly cellulose, were hydrolyzed with cellulase derived from Trichoderma viride. Their enzymatic susceptibility was different among wood species. It was higher in species having lower contents of Klason lignin and guaiacyl lignin. Birch and Mollissima acasia were hydrolyzed more than 90%. Finally, the economics of this process are discussed assuming a plant processing 100 t of hardwoods per day.
Enzyme activity recovery from secondary fiber treated with cellulase and xylanase
1996, Journal of Biotechnology
One of the major problems with implementing biotechnical processes in the recycled paper industry, such as enzyme-enhanced deinking and enzymatic enhancement of pulp drainage properties, is the cost of commercial enzyme preparations. Thus, several factorial studies were performed to determine if enzyme activity can be successfully removed from simulated recycled fiber (once-dried, bleached hardwood and softwood kraft fiber) treated with low concentrations (0.2% or 2.0% on oven-dry fiber) of cellulase or xylanase. Enzyme activity recovery was accomplished by washing treated fiber with dilute NaOH in combination with a low concentration of the nonionic surfactant Tween 80 under a variety of mild conditions. Various cellulase activities i.e., endoglucanase, exoglucanase, and filter paper, can be effectively recovered depending on the washing conditions, the cellulase charge, and the fiber type. Xylanase activity was effectively recovered from softwood, but not hardwood. The results suggest that enzyme activity recovery may be a possible means of decreasing the operating costs for biotechnical processes in the paper industry.
Binding of hemicellulases on isolated polysaccharide substrates
1995, Enzyme and Microbial Technology
Binding of five fungal xylanases produced by Trichoderma reesei, Aspergillus oryzae, and Aspergillus fumigatus and two mannanases of Bacillus subtilis and T. reesei on insoluble xylan, mannan, and cellulose was studied at different pH values and ionic strengths. For comparison, the binding of cellobiohydrolase I (CBH I) and endoglucanase (EG I) of T. reesei on the same substrates was examined. Ionic interactions appeared to play an important role in the binding of xylan, as most of the enzymes were totally bound on xylan when the pH was below their isoelectric point but remained mainly unbound at pH values above the isoelectric point. The binding on xylan was also clearly sensitive to ionic strength. The xylanases were also incompletely bound on mannan and cellulose, and the two mannanases tested were partially bound on mannan. The two cellulolytic enzymes of T. reesei used in this work are both known to contain a cellulose binding domain and were efficiently bound on cellulose. Interestingly, T. reesei mannanase was also found to bind readily on cellulose. Thus, it most probably contains, instead of a mannan binding domain, a specific cellulose binding domain.
Cellulase recycling in biorefineries—is it possible?
2015, Applied Microbiology and Biotechnology

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References (21)

Note on sugar determination

J. Biol. Chem.

Determination of protein: a modification of the Lowry method that gives a linear photometric response

Anal. Biochem.

Analytical separation of Trichoderma reesei cellulases by ion-exchange fast protein chromatography

J. Chromatogr.

Preliminary cost analyses for enzymatic hydrolysis of newsprint

Economic evaluation of enzymatic hydrolysis of phenol-pretreated wheat straw

Biotechnol. Bioeng.

Enzymatic hydrolysis of woods. VII. Enzymatic susceptibility of autohydrolyzed woods

Mokuzai Gakkaishi

Biotechnical utilization of wood carbohydrates after steaming pretreatment

Appl. Microbiol. Biotechnol.

Recycle of enzymes and substrates following enzymatic hydrolysis of steam-pretreated aspenwood

Biotechnol. Bioeng.

Adsorption of cellulase on cellulose: effect of physicochemical properties of cellulose on adsorption and rate of hydrolysis

Biotechnol. Bioeng.

Effect of pore size in substrate and diffusion of enzyme on hydrolysis of cellulosic materials with cellulase

Biotechnol. Bioeng.

Cited by (15)

Valorizing recycled paper sludge by a bioethanol production process with cellulase recycling

Key features of pretreated lignocelluloses biomass solids and their impact on hydrolysis

Integrated process for total utilization of wood components by steam-explosion pretreatment

Enzyme activity recovery from secondary fiber treated with cellulase and xylanase

Binding of hemicellulases on isolated polysaccharide substrates

Cellulase recycling in biorefineries—is it possible?

OriginalAdsorption and desorption of cellulase components during enzymatic hydrolysis of steamed shirakamba (Betula platyphylla Skatchev) wood

Abstract

J. Biol. Chem.

Anal. Biochem.

J. Chromatogr.

Preliminary cost analyses for enzymatic hydrolysis of newsprint

Economic evaluation of enzymatic hydrolysis of phenol-pretreated wheat straw

Biotechnol. Bioeng.

Enzymatic hydrolysis of woods. VII. Enzymatic susceptibility of autohydrolyzed woods

Mokuzai Gakkaishi

Biotechnical utilization of wood carbohydrates after steaming pretreatment

Appl. Microbiol. Biotechnol.

Recycle of enzymes and substrates following enzymatic hydrolysis of steam-pretreated aspenwood

Biotechnol. Bioeng.

Adsorption of cellulase on cellulose: effect of physicochemical properties of cellulose on adsorption and rate of hydrolysis

Biotechnol. Bioeng.

Effect of pore size in substrate and diffusion of enzyme on hydrolysis of cellulosic materials with cellulase

Biotechnol. Bioeng.

Original
Adsorption and desorption of cellulase components during enzymatic hydrolysis of steamed shirakamba (Betula platyphylla Skatchev) wood