1. Investigating the Relationship between Characteristics of Heterogeneous Cellulose and Cellulase Activities
Y-H Percival Zhang, Biological Systems Engineering, Virginia Tech
Blacksburg, VA Tel:
Heterogeneous cellulose accessibility is an important substrate characteristic, but all methods for determining cellulose accessibility to the large-size cellulase molecule have some limitations. Characterization of cellulose accessibility to cellulase (CAC) is vital for better understanding of the enzymatic cellulose hydrolysis mechanism. We have developed a nano-scale tool (recombinant protein) containing a green fluorescent protein and cellulose-binding module. The adsorption of this protein will be used to more accurately represent cellulose accessibility to cellulase than other technologies. Quantitative determination of cellulose accessibility to cellulase (m2/g of cellulose) was established based on the Langmuir adsorption of the fusion protein containing a cellulose-binding module (CBM) and a green fluorescent protein (GFP) (right figures for the protein and adsorption)
Quantitative determinations of the total substrate accessibility to cellulase (TSAC) and cellulose accessibility to cellulase (CAC) based on adsorption of a non-hydrolytic recombinant protein TGC were for the first time established without and with blocking of bovine serum album, respectively. The COSLIF-pretreated corn stover had a CAC of m2/g, nearly twice of that of the dilute acid (DA)-pretreated biomass (5.89 m2/g). All results suggest that CSLF can break lignocellulose structure more efficiently than can DA, resulting in more CAC accompanied by faster hydrolysis rates and higher sugar digestibility.
Langmuir , 23 (25), , Biotechnol. Bioeng. In prep.
Thrombin
Thrombin