Monitoring Cellulase Protein Adsorption and Recovery Using SDS-PAGE

Gang Hu, John A. Heitmann*, Orlando J. Rojas, Joel Pawlak, Dimitris S. Argyropoulos

*Corresponding author for this work

    Research output: Contribution to journalArticleScientificpeer-review

    18 Citations (Scopus)

    Abstract

    Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was employed to study the sorption behaviors of cellulases on microcrystalline celluloses and hardwood pulp. The adsorption and recovery of cellulases from Aspergillus niger and Trichoderma reesei were investigated at 25 degrees C. Cellulase recovery was conducted by rinsing adsorbed enzymes with sodium acetate buffer, Milli-Q water, and sodium hydroxide solution. The initial, equilibrium, and recovered enzymes were analyzed using SDS-PAGE gels. Gels were scanned and analyzed using ImagePro software. The molecular weights of cellulase proteins were determined using a protein marker having seven known proteins. The cellulase system from Trichoderma reesei had a higher adsorption on all substrates studied than the cellulase system from Aspergillius niger, and higher pH favored desorption from the substrates studied. Experimental results also demonstrated that adsorption and desorption amounts determined by SDS-PAGE were proportional to protein concentrations in their crude mixtures.

    Original languageEnglish
    Pages (from-to)8333-8338
    Number of pages6
    JournalIndustrial and Engineering Chemistry Research
    Volume49
    Issue number18
    DOIs
    Publication statusPublished - 15 Sep 2010
    MoE publication typeA1 Journal article-refereed

    Keywords

    • ENZYMATIC-HYDROLYSIS
    • TRICHODERMA-REESEI
    • MECHANISM
    • COMPONENTS
    • ETHANOL
    • RESIDUE
    • VIRIDE
    • ACID
    • WOOD

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