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Aromatic amino acids in the cellulose binding domain of Penicillium crustosum endoglucanase EGL1 differentially contribute to the cellulose affinity of the enzyme
- Source :
- PLoS ONE, PLoS ONE, Vol 12, Iss 5, p e0176444 (2017)
- Publication Year :
- 2016
-
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.
- 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
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 12
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- PloS one
- Accession number :
- edsair.doi.dedup.....8058c681a9ae362d216ccca6f209b504