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Title: Adsorption of Clostridium thermocellum cellulases onto pretreated mixed hardwood, avicel, and lignin

Abstract

Adsorption of Avicel-hydrolyzing activity was examined with respect to: mixed hardwood flour pretreated with 1% sulfuric acid for 9 s at 220C (PTW220), lignin prepared from PTW220 by either acid or enzymatic hydrolysis, and Avicel. Experiments were conducted at 60C for all materials, and also at 25C for PTW220. Based on transient adsorption results and reaction rates, times were selected at which to characterize adsorption at 60C as follows: PTW220, 1 min; lignin, 30 min; and Avicel, 45 min. Similar results were obtained for adsorption of cellulase activity to PTW220 at 25 and 60C, and for lignin prepared by enzymatic and acid hydrolysis. For all materials, adsorption was described well by a Langmuir equation, although the reversibility of adsorption was not investigated. Langmuir affinity constants (L/g) were: PTW220, 109; lignin, 17.9; Avicel, 4.3; cellulose from PTW220, [ge]187. Langmuir capacity constants were 760 for PTW220 and 42 for Avicel; the cellulase binding capacity of lignin appeared to be very high under the conditions examined, and could not be determined. At low and moderate cellulase loadings at least, the majority of cellulase activity adsorbed to PTVV220 is bound to the cellulosic component. The results indicate that PTW220, and its cellulose component inmore » particular, differ radically from Avicel with respect to adsorption. Avicel-hydrolyzing activity and CMC-hydrolyzing activities were found to bind to Avicel with a constant ratio of essentially one, consistent with adsorption of a multi-activity complex.« less

Authors:
; ; ;  [1]
  1. (Dartmouth College, Hanover, NH (United States). Thayer School of Engineering)
Publication Date:
OSTI Identifier:
6204399
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biotechnology and Bioengineering; (United States); Journal Volume: 42:7
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; CELLULASE; ENZYME ACTIVITY; CLOSTRIDIUM THERMOCELLUM; GROWTH; LIGNIN; ENZYMATIC HYDROLYSIS; ADSORPTION; BACTERIA; CARBOHYDRATES; CHEMICAL REACTIONS; CLOSTRIDIUM; DECOMPOSITION; ENZYMES; GLYCOSYL HYDROLASES; HYDROLASES; HYDROLYSIS; LYSIS; MICROORGANISMS; O-GLYCOSYL HYDROLASES; ORGANIC COMPOUNDS; POLYSACCHARIDES; PROTEINS; SACCHARIDES; SOLVOLYSIS; SORPTION; 090900* - Biomass Fuels- Processing- (1990-)

Citation Formats

Bernardez, T.D., Lyford, K., Hogsett, D.A., and Lynd, L.R.. Adsorption of Clostridium thermocellum cellulases onto pretreated mixed hardwood, avicel, and lignin. United States: N. p., 1993. Web. doi:10.1002/bit.260420715.
Bernardez, T.D., Lyford, K., Hogsett, D.A., & Lynd, L.R.. Adsorption of Clostridium thermocellum cellulases onto pretreated mixed hardwood, avicel, and lignin. United States. doi:10.1002/bit.260420715.
Bernardez, T.D., Lyford, K., Hogsett, D.A., and Lynd, L.R.. Mon . "Adsorption of Clostridium thermocellum cellulases onto pretreated mixed hardwood, avicel, and lignin". United States. doi:10.1002/bit.260420715.
@article{osti_6204399,
title = {Adsorption of Clostridium thermocellum cellulases onto pretreated mixed hardwood, avicel, and lignin},
author = {Bernardez, T.D. and Lyford, K. and Hogsett, D.A. and Lynd, L.R.},
abstractNote = {Adsorption of Avicel-hydrolyzing activity was examined with respect to: mixed hardwood flour pretreated with 1% sulfuric acid for 9 s at 220C (PTW220), lignin prepared from PTW220 by either acid or enzymatic hydrolysis, and Avicel. Experiments were conducted at 60C for all materials, and also at 25C for PTW220. Based on transient adsorption results and reaction rates, times were selected at which to characterize adsorption at 60C as follows: PTW220, 1 min; lignin, 30 min; and Avicel, 45 min. Similar results were obtained for adsorption of cellulase activity to PTW220 at 25 and 60C, and for lignin prepared by enzymatic and acid hydrolysis. For all materials, adsorption was described well by a Langmuir equation, although the reversibility of adsorption was not investigated. Langmuir affinity constants (L/g) were: PTW220, 109; lignin, 17.9; Avicel, 4.3; cellulose from PTW220, [ge]187. Langmuir capacity constants were 760 for PTW220 and 42 for Avicel; the cellulase binding capacity of lignin appeared to be very high under the conditions examined, and could not be determined. At low and moderate cellulase loadings at least, the majority of cellulase activity adsorbed to PTVV220 is bound to the cellulosic component. The results indicate that PTW220, and its cellulose component in particular, differ radically from Avicel with respect to adsorption. Avicel-hydrolyzing activity and CMC-hydrolyzing activities were found to bind to Avicel with a constant ratio of essentially one, consistent with adsorption of a multi-activity complex.},
doi = {10.1002/bit.260420715},
journal = {Biotechnology and Bioengineering; (United States)},
number = ,
volume = 42:7,
place = {United States},
year = {Mon Sep 20 00:00:00 EDT 1993},
month = {Mon Sep 20 00:00:00 EDT 1993}
}
  • The cellulase activity in cell-free broths from Clostridium thermocellum is examined on both dilute-acid-pretreated mixed hardwood (90% maple, 10% birch) and Avicel. Experiments were conducted in vitro in order to distinguish properties of the cellulase from properties of the organism and to evaluate the effectiveness of C. thermocellum cellulase in the hydrolysis of a naturally occurring, lignin-containing substrate. The results obtained establish that essentially quantitative hydrolysis of cellulose from pretreated mixed hardwood is possible using this enzyme system. Pretreatment with 1% H/sub 2/SO/sub 4/ and a 9-s residence time at 220, 210, 200, and 180/sup 0/C allowed yields after enzymaticmore » hydrolysis (percentage of glucan solubilized/glucan potentially solubilized) of 97.8, 86.1, 82.0, and 34.6%, respectively. Enzymatic hydrolysis of mixed hardwood with no pretreatment resulted in a yield of 10.1%. Hydrolysis yields of greater than 95% were obtained from 0.6 g/l mixed hardwood pretreated at 220/sup 0/C in 7 hours at broth strengths of 60 and 80% (v/v) and in approximately 48 hours with 33% broth. Hydrolysis of pretreated mixed hardwood is compared to hydrolysis of Avicel. The initial rate of Avicel hydrolysis saturates with respect to enzyme, whereas the initial rate of hydrolysis of pretreated wood is proportional to the amount of enzyme present. Initial hydrolysis rates for pretreated wood and Avicel at 0.6 g/l are greater for wood at low broth dilutions (1.25:1 to 5:1) by up to 2.7-fold and greater for Avicel at high broth dilutions (5:1 to 50:1) by up to 4.3-fold. Maximum rates of hydrolysis are achieved at less than 2 g substrate/liter for both pretreated wood and Avicel).« less
  • Broussonetia papyrifera, known as paper mulberry, is a potential feed stock for bioethanol production because of its cellulose-rich composition. Lignin in dilute acid pretreated Broussonetia papyrifera was fractionated to three different fractions, and their physiochemical properties were determined by FT-IR, GPC and NMR analyses. Different structural characteristics were observed from each lignin fraction. Cellobiohydrolases I (CBH) adsorption to each lignin was understood by the lignin properties. The results showed that aliphatic hydroxyl groups in lignin showed positive correlations with the maximum binding ability of CBH onto lignin samples. Also, the contents of phenolic compounds such as p-hydroxyphenyl benzoate (PB), syringylmore » (S) and guaiacyl (G) units in the lignin influenced their CBH binding.« less
  • Here, to develop and prototype a high-throughput microplate assay to assess anaerobic microorganisms and lignocellulosic biomasses in a rapid, cost-effective screen for consolidated bioprocessing potential. Clostridium thermocellum parent Δ hpt strain deconstructed Avicel to cellobiose, glucose, and generated lactic acid, formic acid, acetic acid and ethanol as fermentation products in titers and ratios similar to larger scale fermentations confirming the suitability of a plate-based method for C. thermocellum growth studies. C. thermocellum strain LL1210, with gene deletions in the key central metabolic pathways, produced higher ethanol titers in the Consolidated Bioprocessing (CBP) plate assay for both Avicel and switchgrass fermentationsmore » when compared to the Δ hpt strain. A prototype microplate assay system is developed that will facilitate high-throughput bioprospecting for new lignocellulosic biomass types, genetic variants and new microbial strains for bioethanol production.« less