Design of highly efficient cellulase mixtures for enzymatic hydrolysis of cellulose

An extremely highly active cellobiohydrolase (CBH IIb or Cel6B) was isolated from Chrysosporium lucknowense UV18‐25 culture filtrate. The CBH IIb demonstrated the highest ability for a deep degradation of crystalline cellulose amongst a few cellobiohydrolases tested, including C. lucknowense CBH Ia, Ib, IIa, and Trichoderma reesei CBH I and II. Using purified C. lucknowense enzymes (CBH Ia, Ib, and IIb; endoglucanases II and V; β‐glucosidase, xylanase II), artificial multienzyme mixtures were reconstituted, displaying an extremely high performance in a conversion of different cellulosic substrates (Avicel, cotton, pretreated Douglas fir wood) to glucose. These mixtures were much or notably more effective in hydrolysis of the cellulosic substrates than the crude multienzyme C. lucknowense preparation and other crude cellulase samples produced by T. reesei and Penicillium verruculosum. Highly active cellulases are a key factor in bioconversion of plant lignocellulosic biomass to ethanol as an alternative to fossil fuels. Biotechnol. Bioeng. 2007; 97: 1028–1038. © 2007 Wiley Periodicals, Inc.

[1]  B Henrissat,et al.  A classification of glycosyl hydrolases based on amino acid sequence similarities. , 1991, The Biochemical journal.

[2]  Gusakov,et al.  Surface hydrophobic amino acid residues in cellulase molecules as a structural factor responsible for their high denim-washing performance. , 2000, Enzyme and microbial technology.

[3]  R. Blanchette,et al.  Biodegradation of Cellulose , 1990 .

[4]  A. Kurabi,et al.  Enzymatic hydrolysis of steam-exploded and ethanol organosolv-pretreated douglas-fir by novel and commercial fungal cellulases , 2005, Applied biochemistry and biotechnology.

[5]  G. Pettersson,et al.  The initial kinetics of hydrolysis by cellobiohydrolases I and II is consistent with a cellulose surface-erosion model. , 1998, European journal of biochemistry.

[6]  S L Mowbray,et al.  Family 7 cellobiohydrolases from Phanerochaete chrysosporium: crystal structure of the catalytic module of Cel7D (CBH58) at 1.32 A resolution and homology models of the isozymes. , 2001, Journal of molecular biology.

[7]  Stephen E. Wald,et al.  Kinetics of the enzymatic hydrolysis of cellulose , 1984, Biotechnology and bioengineering.

[8]  W. D. Murray,et al.  Bioconversion of forest products industry waste cellulosics to fuel ethanol: a review. , 1996 .

[9]  Alexander V. Gusakov,et al.  Purification, cloning and characterisation of two forms of thermostable and highly active cellobiohydrolase I (Cel7A) produced by the industrial strain of , 2005 .

[10]  A. Gusakov,et al.  Enzymatic saccharification of industrial and agricultural lignocellulosic wastes , 1992 .

[11]  L. Olsson,et al.  Fermentation of lignocellulosic hydrolysates for ethanol production. , 1996 .

[12]  W. Steiner,et al.  Cellulose hydrolysis by the cellulases from Trichoderma reesei: a new model for synergistic interaction. , 1994, The Biochemical journal.

[13]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[14]  M. Schülein Enzymatic properties of cellulases from Humicola insolens. , 1997, Journal of biotechnology.

[15]  T. V. Bubnova,et al.  The selection and properties of Penicillium verruculosum mutants with enhanced production of cellulases and xylanases , 2005, Microbiology.

[16]  B. Ganem,et al.  Identification of two functionally different classes of exocellulases. , 1996, Biochemistry.

[17]  Richard T. Elander,et al.  Survey and analysis of commercial cellulase preparations suitable for biomass conversion to ethanol , 1997 .

[18]  A Bairoch,et al.  Updating the sequence-based classification of glycosyl hydrolases. , 1996, The Biochemical journal.

[19]  Dr. Peter James Proteome Research: Mass Spectrometry , 2001, Principles and Practice.

[20]  D. Kilburn,et al.  Evaluation of novel fungal cellulase preparations for ability to hydrolyze softwood substrates – evidence for the role of accessory enzymes , 2005 .

[21]  Michael Somogyi,et al.  NOTES ON SUGAR DETERMINATION , 1926 .

[22]  A. Gusakov,et al.  Kinetics of the enzymatic hydrolysis of cellulose: 1. A mathematical model for a batch reactor process , 1985 .

[23]  B. Henrissat,et al.  Imaging the Enzymatic Digestion of Bacterial Cellulose Ribbons Reveals the Endo Character of the Cellobiohydrolase Cel6A from Humicola insolens and Its Mode of Synergy with Cellobiohydrolase Cel7A , 2000, Applied and Environmental Microbiology.

[24]  Johan Karlsson,et al.  A model explaining declining rate in hydrolysis of lignocellulose substrates with cellobiohydrolase I (Cel7A) and endoglucanase I (Cel7B) of Trichoderma reesei , 2002, Applied biochemistry and biotechnology.

[25]  Tuula T. Teeri,et al.  Crystalline cellulose degradation : new insight into the function of cellobiohydrolases , 1997 .

[26]  B. Henrissat,et al.  Synergism of Cellulases from Trichoderma reesei in the Degradation of Cellulose , 1985, Bio/Technology.

[27]  Maobing Tu,et al.  Weak lignin-binding enzymes , 2005, Applied biochemistry and biotechnology.

[28]  A. I. Antonov,et al.  Cellulase Complex of the Fungus Chrysosporium lucknowense: Isolation and Characterization of Endoglucanases and Cellobiohydrolases , 2004, Biochemistry (Moscow).

[29]  G. Tiraby,et al.  Genetic improvement of Trichoderma reesei for large scale cellulase production , 1988 .

[30]  Himmel,et al.  Cellulase for commodity products from cellulosic biomass , 1999, Current opinion in biotechnology.

[31]  T. K. Ghose Measurement of cellulase activities , 1987 .

[32]  Henry R. Bungay,et al.  Energy, the biomass options , 1981 .

[33]  J. Ståhlberg,et al.  Adsorption and synergism of cellobiohydrolase I and II of Trichoderma reesei during hydrolysis of microcrystalline cellulose , 1994, Biotechnology and bioengineering.

[34]  J. Pérez,et al.  Biodegradation and biological treatments of cellulose, hemicellulose and lignin: an overview , 2002, International microbiology : the official journal of the Spanish Society for Microbiology.

[35]  T. Houfek,et al.  Transcriptional Regulation of Biomass-degrading Enzymes in the Filamentous Fungus Trichoderma reesei* , 2003, Journal of Biological Chemistry.

[36]  T. Teeri,et al.  The Cellulases Endoglucanase I and Cellobiohydrolase II of Trichoderma reesei Act Synergistically To Solubilize Native Cotton Cellulose but Not To Decrease Its Molecular Size , 1996, Applied and environmental microbiology.

[37]  J. Nielsen,et al.  Fuel ethanol production from lignocellulose: a challenge for metabolic engineering and process integration , 2001, Applied Microbiology and Biotechnology.

[38]  A. Sinitsyn,et al.  Comparative evaluation of hydrolytic efficiency toward microcrystalline cellulose of Penicillium and Trichoderma cellulases , 1995 .

[39]  M. Galbe,et al.  A review of the production of ethanol from softwood , 2002, Applied Microbiology and Biotechnology.

[40]  A. Clarke Biodegradation of Cellulose: Enzymology and Biotechnology , 1996 .

[41]  T. Wood,et al.  Synergism Between Enzymes Involved in the Solubilization of Native Cellulose , 1979 .

[42]  S. Covert,et al.  Structure, organization, and transcription of a cellobiohydrolase gene cluster from Phanerochaete chrysosporium , 1992, Applied and environmental microbiology.

[43]  Weak lignin-binding enzymes , 2005 .