A cellulose-binding module of the Trichoderma reesei @b-mannanase Man5A increases the mannan-hydrolysis of complex substrates

Per Hägglund, Torny Eriksson, Anna Collén, Wim Nerinckx, Marc Claeyssens, Henrik Stålbrand

Research output: Contribution to journalArticlepeer-review

77 Citations (SciVal)

Abstract

Endo-@b-1,4-d-mannanases (@b-mannanase; EC 3.2.1.78) are endohydrolases that participate in the degradation of hemicellulose, which is closely associated with cellulose in plant cell walls. The @b-mannanase from Trichoderma reesei (Man5A) is composed of an N-terminal catalytic module and a C-terminal carbohydrate-binding module (CBM). In order to study the properties of the CBM, a construct encoding a mutant of Man5A lacking the part encoding the CBM (Man5A@DCBM), was expressed in T. reesei under the regulation of the Aspergillus nidulans gpdA promoter. The wild-type enzyme was expressed in the same way and both proteins were purified to electrophoretic homogeneity using ion-exchange chromatography. Both enzymes hydrolysed mannopentaose, soluble locust bean gum galactomannan and insoluble ivory nut mannan with similar rates. With a mannan/cellulose complex, however, the deletion mutant lacking the CBM showed a significant decrease in hydrolysis. Binding experiments using activity detection of Man5A and Man5A@DCBM suggests that the CBM binds to cellulose but not to mannan. Moreover, the binding of Man5A to cellulose was compared with that of an endoglucanase (Cel7B) from T. reesei.
Original languageEnglish
Pages (from-to)37-48
JournalJournal of Biotechnology
Volume101
Issue number1
DOIs
Publication statusPublished - 2003

Subject classification (UKÄ)

  • Biological Sciences

Keywords

  • Carbohydrate-binding module
  • Hemicellulase
  • Cellulose
  • Hemicellulose
  • Endoglucanase

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