Synthetic multi-component enzyme mixtures for deconstruction of lignocellulosic biomass

A high throughput enzyme assay platform, called GENPLAT, was used to guide the development of an optimized mixture of individual purified enzymes from ten “accessory” and six “core” enzymes. Enzyme mixtures were optimized for release of Glu, Xyl, or a combination of the two from corn stover pretreat...

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Bibliographic Details
Published in:Bioresource technology 2010-12, Vol.101 (23), p.9097-9105
Main Authors: Banerjee, Goutami, Car, Suzana, Scott-Craig, John S., Borrusch, Melissa S., Bongers, Mareike, Walton, Jonathan D.
Format: Article
Language:English
Subjects:
Ammonia - metabolism
Ammonia-fiber expansion
Biological and medical sciences
Biomass
Calorimetry
Cellulase
Chromatography, Gas
Electrophoresis, Polyacrylamide Gel
Enzyme Assays
Enzymes - metabolism
Fundamental and applied biological sciences. Psychology
Fungal Proteins - metabolism
Glucose - metabolism
Kinetics
Lignin - metabolism
Lignocellulose
Pichia - enzymology
Pichia pastoris
Time Factors
Trichoderma - enzymology
Trichoderma reesei
Xylose - metabolism
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Summary:A high throughput enzyme assay platform, called GENPLAT, was used to guide the development of an optimized mixture of individual purified enzymes from ten “accessory” and six “core” enzymes. Enzyme mixtures were optimized for release of Glu, Xyl, or a combination of the two from corn stover pretreated by ammonia-fiber expansion (AFEX). Assay conditions were a fixed enzyme loading of 15 mg/g glucan, 48 h digestion, and 50 °C. Five of the ten tested accessory proteins enhanced Glu or Xyl yield compared to the core set alone, and five did not. An 11-component mixture containing the core set and five accessory enzymes optimized for Glu released 52.1% of the available Glu, compared to 38.5% with the core set alone. A mixture optimized for Xyl released 39.9% of the Xyl, compared to 26.4% with the core set alone. We predict that there is still considerable opportunity for further improvement of synthetic mixtures. Furthermore, the strategy described here is applicable to the development of more efficient enzyme cocktails for any pretreatment/biomass combination and for detecting enzymes that make a heretofore unrecognized contribution to lignocellulose deconstruction.
ISSN:0960-8524
1873-2976
DOI:10.1016/j.biortech.2010.07.028