title: Purification and biochemical characterization of glucose-cellobiose-tolerant cellulases from Scytalidium thermophilum.
abstract: Two cellulases from Scytalidium thermophilum were purified and characterized, exhibiting tolerance to glucose and cellobiose. Characterization of purified cellulases I and II by mass spectrometry revealed primary structure similarities with an exoglucanase and an endoglucanase, respectively. Molecular masses were 51.2 and 45.6 kDa for cellulases I and II, respectively, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Cellulases I and II exhibited isoelectric points of 6.2 and 6.9 and saccharide contents of 11 and 93 %, respectively. Optima of temperature and pH were 60-65 °C and 4.0 for purified cellulase I and 65 °C and 6.5 for purified cellulase II. Both cellulases maintained total CMCase activity after 60 min at 60 °C. Cysteine, Mn(2+), dithiotreitol and ß-mercaptoethanol-stimulated cellulases I and II. The tolerance to cellulose hydrolysis products and the high thermal stabilities of Scytalidium cellulases suggest good potential for industrial applications.
authors.affiliation: Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Jean Carlos Rodrigues
authors.initials: JC
authors.lastname: Silva
authors.forename: Luis Henrique Souza
authors.initials: LH
authors.lastname: Guimarães
authors.forename: José Carlos Santos
authors.initials: JC
authors.lastname: Salgado
authors.forename: Rosa Prazeres Melo
authors.initials: RP
authors.lastname: Furriel
authors.forename: Maria Lourdes T M
authors.initials: ML
authors.lastname: Polizeli
authors.forename: José César
authors.initials: JC
authors.lastname: Rosa
authors.forename: João Atilio
authors.initials: JA
authors.lastname: Jorge
mesh_terms: D001203:Ascomycota / Q000201:enzymology*; D002475:Cellobiose / Q000378:metabolism*; D044602:Cellulases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D020536:Enzyme Activators / Q000032:analysis; D004791:Enzyme Inhibitors / Q000378:metabolism*; D004795:Enzyme Stability; D005947:Glucose / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D013058:Mass Spectrometry; D008970:Molecular Weight; D013696:Temperature
chemical_list: D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D002475:Cellobiose; D044602:Cellulases; D005947:Glucose
references.citation: Protein Sci. 2003 Dec;12(12):2782-93
references.pmid: 14627738
references.citation: J Agric Food Chem. 2010 Sep 8;58(17):9833-7
references.pmid: 20687562
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: Cell. 1977 Dec;12(4):1133-41
references.pmid: 23215
references.citation: J Agric Food Chem. 2010 May 26;58(10):6126-30
references.pmid: 20415423
references.citation: J Microbiol. 2010 Feb;48(1):53-62
references.pmid: 20221730
references.citation: Microbiol Mol Biol Rev. 2000 Sep;64(3):461-88
references.pmid: 10974122
references.citation: Bioresour Technol. 2003 Aug;89(1):17-34
references.pmid: 12676497
references.citation: J Ind Microbiol Biotechnol. 2008 Jun;35(6):515-24
references.pmid: 18228067
references.citation: Biochem Mol Biol Educ. 2005 Nov;33(6):399-403
references.pmid: 21638609
references.citation: Trends Biotechnol. 2006 Dec;24(12):549-56
references.pmid: 17050014
references.citation: Biotechnol Adv. 1997;15(3-4):583-620
references.pmid: 14538158
references.citation: J Biol Chem. 1951 Nov;193(1):265-75
references.pmid: 14907713
references.citation: Folia Microbiol (Praha). 1997;42(4):303-11
references.pmid: 9449776
references.citation: Biotechnol Bioeng. 2007 Jun 1;97(2):287-96
references.pmid: 17058283
references.citation: Adv Biochem Eng Biotechnol. 2007;108:121-45
references.pmid: 17589813
references.citation: J Ind Microbiol Biotechnol. 2008 May;35(5):377-391
references.pmid: 18338189