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Proteins, Expression, Isolation and Analysis> | … Protein from the SDS-PAGE gel was transferred to nitrocellulose membrane using the eBlot Protein Transfer System (GenScript, USA) The membrane was blocked by 5% (w/v) skimmed milk in TBST over night at room temperature, then washed three times with 10 mL TBST … | Get A Quote |
L-Arabinose isomerase isolated from Geobacillus stearothermophilus (GSAI) was modified to improve its substrate specificity for D-galactose for the production of D-tagatose, a potential reduced-energy sweetener. Among the selected residues, mutation at residue 18 produced a mutant strain, H18T, which exhibited increased activity for D-galactose compared with the wild-type (WT) enzyme. Analysis of the substrate specificity of H18T showed a 45.4% improvement for D-galactose. Replacing histidine with threonine at residue 18 resulted in approximately 2.7-fold and 1.8-fold higher substrate binding and catalytic efficiency, respectively, for D-galactose. Further enhancement of the specific activity and catalytic effi... More