Implications of Cellobiohydrolase Glycosylation for use in Biomass Conversion

Tina Jeoh, William Michener, Michael Himmel, Steven Decker, William Adney

Research output: Contribution to journalArticlepeer-review


The cellulase producing ascomycete, Trichoderma reesei (Hypocrea jecorina), is known to secrete a range of enzymes important for ethanol production from lignocellulosic biomass. It is also widely used for the commercial scale production of industrial enzymes because of its ability to produce high titers of heterologous proteins. During the secretion process, a number of post-translational events can occur, however, that impact protein function and stability. Another ascomycete, Aspergillus niger var. awamori, is also known to produce large quantities of heterologous proteins for industry. In this study, T. reesei Cel7A, a cellobiohydrolase, was expressed in A. niger var. awamori and subjected to detailed biophysical characterization. The purified recombinant enzyme contains six times the amount of N-linked glycan than the enzyme purified from a commercial T. reesei enzyme preparation. The activities of the two enzyme forms were compared using bacterial (microcrystalline) and phosphoric acid swollen (amorphous) cellulose as substrates. This comparison suggested that the increased level of N-glycosylation of the recombinant Cel7A (rCel7A) resulted in reduced activity and increased non-productive binding on cellulose. When treated with the N-glycosidase PNGaseF, the molecular weight of the recombinant enzyme approached that of the commercial enzyme and the activity on cellulose was improved.
Original languageAmerican English
Pages (from-to)10-22
Number of pages13
JournalBiotechnology for Biofuels
Issue number10
StatePublished - 2008

NREL Publication Number

  • NREL/JA-270-42587


  • bacterial cellulose
  • catalytic domain
  • cellulase
  • cellulose conversion
  • glycosyl hydrolase


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