Glutantbase

GlutantBase facilitates rational design of glucose-tolerant β-glucosidases by compiling GH1 β-glucosidase sequences and integrating structural, mutational, and docking analyses to guide engineering for improved glucose tolerance and thermostability in lignocellulose saccharification.

Key Features:

• Extensive sequence database: Contains 3,842 GH1 β-glucosidase sequences sourced from UniProt for comparative sequence analysis.
• Homology modeling and structural prediction: Predicts three-dimensional structures and highlights structural features such as catalytic pockets, conserved amino acids, and secondary structure elements.
• Catalytic and functional insights: Reports catalytic residues, coevolution networks, and channel residues that guide substrates to the active site.
• Mutation analysis and prediction: Analyzes beneficial mutations reported in the literature and predicts analogous mutations based on six characterized mutants (A404V, E96K, H184F, H228T, L441F, V174C) associated with enhanced catalytic activity, glucose tolerance, or thermostability.
• Molecular docking studies: Uses molecular docking to assess the impact of suggested mutations on enzyme–ligand interactions, including identification of H228T-based mutations that reduce glucose affinity while increasing cellobiose affinity.

Scientific Applications:

• Rational enzyme engineering for lignocellulose saccharification: Supports design strategies to mitigate product inhibition by glucose during cellobiose hydrolysis.
• Prioritization of mutations for experimental validation: Guides selection of candidate mutations to improve glucose tolerance and thermostability based on comparative and docking analyses.
• Structure–function analysis: Enables investigation of catalytic residues, coevolution networks, and channel architecture to interpret determinants of enzyme specificity and stability.

Methodology:

Sequence collection from UniProt (3,842 GH1 β-glucosidase sequences), homology modeling for 3D structure prediction, identification of catalytic residues, coevolution network and channel residue analyses, literature-based mutation analysis of six mutants (A404V, E96K, H184F, H228T, L441F, V174C), and molecular docking to assess mutation effects on enzyme–ligand interactions.