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Biochemistry (Moscow)

, Volume 69, Issue 5, pp 542–551 | Cite as

Cellulase Complex of the Fungus Chrysosporium lucknowense: Isolation and Characterization of Endoglucanases and Cellobiohydrolases

  • F. E. Bukhtojarov
  • B. B. Ustinov
  • T. N. Salanovich
  • A. I. Antonov
  • A. V. GusakovEmail author
  • O. N. Okunev
  • A. P. Sinitsyn
Article

Abstract

Using different chromatographic techniques, eight cellulolytic enzymes were isolated from the culture broth of a mutant strain of Chrysosporium lucknowense: six endoglucanases (EG: 25 kD, pI 4.0; 28 kD, pI 5.7; 44 kD, pI 6.0; 47 kD, pI 5.7; 51 kD, pI 4.8; 60 kD, pI 3.7) and two cellobiohydrolases (CBH I, 65 kD, pI 4.5; CBH II, 42 kD, pI 4.2). Some of the isolated cellulases were classified into known families of glycoside hydrolases: Cel6A (CBH II), Cel7A (CBH I), Cel12A (EG28), Cel45A (EG25). It was shown that EG44 and EG51 are two different forms of one enzyme. EG44 seems to be a catalytic module of an intact EG51 without a cellulose-binding module. All the enzymes had pH optimum of activity in the acidic range (at pH4.5-6.0), whereas EG25 and EG47 retained 55-60% of the maximum activity at pH 8.5. Substrate specificity of the purified cellulases against carboxymethylcellulose (CMC), β-glucan, Avicel, xylan, xyloglucan, laminarin, and p-nitrophenyl-β-D-cellobioside was studied. EG44 and EG51 were characterized by the highest CMCase activity (59 and 52 U/mg protein). EG28 had the lowest CMCase activity (11 U/mg) amongst the endoglucanases; however, this enzyme displayed the highest activity against β-glucan (125 U/mg). Only EG51 and CBH I were characterized by high adsorption ability on Avicel cellulose (98-99%). Kinetics of Avicel hydrolysis by the isolated cellulases in the presence of purified β-glucosidase from Aspergillus japonicus was studied. The hydrolytic efficiency of cellulases (estimated as glucose yield after a 7-day reaction) decreased in the following order: CBH I, EG60, CBH II, EG51, EG47, EG25, EG28, EG44.

cellulase endoglucanase cellobiohydrolase chromatography adsorption saccharification 

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Copyright information

© MAIK “Nauka/Interperiodica” 2004

Authors and Affiliations

  • F. E. Bukhtojarov
    • 1
  • B. B. Ustinov
    • 1
  • T. N. Salanovich
    • 1
  • A. I. Antonov
    • 1
  • A. V. Gusakov
    • 1
    Email author
  • O. N. Okunev
    • 2
  • A. P. Sinitsyn
    • 1
  1. 1.Faculty of ChemistryLomonosov Moscow State UniversityMoscowRussia
  2. 2.Institute of Biochemistry and Physiology of MicroorganismsRussian Academy of Sciences, PushchinoMoscow RegionRussia

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