Abstract
Although a poor indicator of how a cellulase preparation will perform on biomass, the filter paper unit (FPU) still finds wide use in the literature as an apparent measure of performance efficacy. In actuality, the assessment of commercial enzyme preparation performance in terms of biomass conversion or solubilization of insoluble polysaccharides is largely dependent on the substrate composition, which cannot be easily standardized. Commercial cellulase preparations are evaluated based upon their performance or specific activity. The ability to compare commercial enzyme preparation efficacy across a wide variety of different preparations requires defining the amount of enzyme protein required in milligrams per gram of cellulose to achieve a targeted level of cellulose hydrolysis in a specified timeframe. Since biomass substrates are highly variable, reproducible and accurate protein determination is as important as performance testing to be able to rank order the effectiveness of diverse preparations. This chapter describes a protocol that overcomes many of the difficulties encountered with determining the protein concentration in commercial cellulase preparations.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Aden A, Foust T (2009) Technoeconomic analysis of the dilute sulfuric acid and enzymatic hydrolysis process for the conversion of corn stover to ethanol. Cellulose 16:535–545
Merino ST, Cherry J (2007) Progress and challenges in enzyme development for biomass utilization. Adv Biochem Eng Biotechnol 108:95–120
Teter SA, Xu F, Nedwin GE, Cherry JR (2006) Enzymes for biorefineries. In: Kamm B, Gruber PR, Kamm M (eds) Biorefineries - industrial processes and products, vol 1. Wiley-VCH, Weinheim, pp 357–383
Dean B, Dodge T, Valle F, Chotani G (2006) Development of biorefineries - technical and economic considerations. In: Kamm B, Gruber PR (eds) Biorefineries - industrial processes and products, vol 1. Wiley-VCH, Weinheim, pp 67–83
Lynd LR, Laser MS, Brandsby D, Dale BE, Davison B, Hamilton R, Himmel M, Keller M, McMillan JD, Sheehan J, Wyman CE (2008) How biotech can transform biofuels. Nat Biotechnol 26:169–172
Himmel ME (2009) Corn stover conversion to biofuels: DOE’s preparation for readiness in 2012. Cellulose 16:531–534
Wilson DB (2009) Cellulases and biofuels. Curr Opin Biotechnol 20:295–299
Tolan JS, Foody B (1999) Cellulase from submerged fermentation. Adv Biochem Eng/Biotechnol 65:41–67
Tolan JS (2006) Iogen’s demonstration process for producing ethanol from cellulosic biomass. In: Kamm B, Gruber PR (eds) Biorefineries - industrial processes and products, vol 1. Wiley-VCH, Weinhelm, pp 193–208, 2 vols
Cen P (1999) Production of cellulase by solid-state fermentation. Adv Biochem Eng/Biotechnol 65:69–92
Elander RT, Dale BE, Holtzapple M, Ladisch MR, Lee YY, Mitchinson C, Saddler JN, Wyman CE (2009) Summary of findings from the Biomass Refining Consortium for Applied Fundamentals and Innovation (CAFI): corn stover pretreatment. Cellulose 16:649–659
Thermo Fisher Scientific I (2010) Thermo Scientific Pierce Protein Assay Technical Handbook, Version 2
Ghose GL (1987) Measurement of cellulase activities. Pure Appl Chem 59:257–268
Nieves RA, Ehrman CI, Adney WS, Elander RT, Himmel ME (1998) Survey and analysis of commercial cellulase preparations suitable for biomass conversion to ethanol. World J Microbiol Biotechnol 14:301–304
Timasheff SN (1998) Control of protein stability and reactions by weakly interacting cosolvents: the simplicity of the complicated. Adv Protein Chem 51:355–432
Vagenende V, Yap MGS, Trout BL (2009) Mechanisms of protein stabilization and prevention of protein aggregation by glycerol. Biochemistry 48:11084–11096
Adney William S, Mohagheghi A, Thomas Steven R, Himmel Michael E (1996) Comparison of protein contents of cellulase preparations in a worldwide Round-Robin assay. In: Enzymatic degradation of insoluble carbohydrates, vol 618, pp 256–271. American Chemical Society
Adney WS, Taylor LE, Johnson D, Park S, Knoshaug EP, Nimlos MR, Decker SR, Vinzant TB, Selig MJ, Himmel ME (2008) CELL 215-Deconstruction of biomass: Understanding enzyme/substrate interactions. Abstracts of Papers of the American Chemical Society. p 235
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Adney, W.S., Dowe, N., Jennings, E.W., Mohagheghi, A., Yarbrough, J., McMillan, J.D. (2012). Assessing the Protein Concentration in Commercial Enzyme Preparations. In: Himmel, M. (eds) Biomass Conversion. Methods in Molecular Biology, vol 908. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-956-3_16
Download citation
DOI: https://doi.org/10.1007/978-1-61779-956-3_16
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-955-6
Online ISBN: 978-1-61779-956-3
eBook Packages: Springer Protocols