151. Aromatic amino acids in the cellulose binding domain of Penicillium crustosum endoglucanase EGL1 differentially contribute to the cellulose affinity of the enzyme
- Author
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Zheng-gang Han, Jiang-Ke Yang, Fang-Yuan Chen, Li Xu, and Wei Xiong
- Subjects
0301 basic medicine ,lcsh:Medicine ,Pathology and Laboratory Medicine ,Biochemistry ,Substrate Specificity ,chemistry.chemical_compound ,Binding Analysis ,Aromatic Amino Acids ,Filter Paper ,Aromatic amino acids ,Medicine and Health Sciences ,Cellulases ,Amino Acids ,lcsh:Science ,chemistry.chemical_classification ,Multidisciplinary ,biology ,Organic Compounds ,Amino acid ,Enzymes ,Laboratory Equipment ,Chemistry ,Physical Sciences ,Engineering and Technology ,Cell Binding Assay ,Research Article ,Cell Binding ,Cell Physiology ,Green Fluorescent Proteins ,Equipment ,Cellulase ,Calorimetry ,Research and Analysis Methods ,03 medical and health sciences ,Amino Acids, Aromatic ,Signs and Symptoms ,Diagnostic Medicine ,Enzymatic hydrolysis ,Cellulose ,Penicillium crustosum ,Chemical Characterization ,030102 biochemistry & molecular biology ,Facies (Medical) ,lcsh:R ,Organic Chemistry ,Chemical Compounds ,Biology and Life Sciences ,Proteins ,Isothermal titration calorimetry ,Cell Biology ,biology.organism_classification ,Cellulose binding ,digestive system diseases ,030104 developmental biology ,chemistry ,biology.protein ,Enzymology ,lcsh:Q - Abstract
The cellulose binding domain (CBD) of cellulase binding to cellulosic materials is the initiation of a synergistic action on the enzymatic hydrolysis of the most abundant renewable biomass resources in nature. The binding of the CBD domain to cellulosic substrates generally relies on the interaction between the aromatic amino acids structurally located on the flat face of the CBD domain and the glucose rings of cellulose. In this study, we found the CBD domain of a newly cloned Penicillium crustosum endoglucanase EGL1, which was phylogenetically related to Aspergillus, Fusarium and Rhizopus, and divergent from the well-characterized Trichoderma reeseis cellulase CBD domain, contain two conserved aromatic amino acid-rich regions, Y451-Y452 and Y477-Y478-Y479, among which three amino acids Y451, Y477, and Y478 structurally sited on a flat face of this domain. Cellulose binding assays with green fluorescence protein as the marker, adsorption isotherm assays and an isothermal titration calorimetry assays revealed that although these three amino acids participated in this process, the Y451-Y452 appears to contribute more to the cellulose binding than Y477-Y478-Y479. Further glycine scanning mutagenesis and structural modelling revealed that the binding between CBD domain and cellulosic materials might be multi-amino-acids that participated in this process. The flexible poly-glucose molecule could contact Y451, Y477, and Y478 which form the contacting flat face of CBD domain as the typical model, some other amino acids in or outside the flat face might also participate in the interaction. Thus, it is possible that the conserved Y451-Y452 of CBD might have a higher chance of contacting the cellulosic substrates, contributing more to the affinity of CBD than the other amino acids.
- Published
- 2016