title: Influence of Buffer Composition and Calcium Chloride on GdnHCl Denaturation of Bacillus licheniformis α-Amylase.
abstract: The influence of buffer composition on the conformational stability of native and calciumdepleted Bacillus licheniformis α-amylase (BLA) was investigated against guanidine hydrochloride (GdnHCl) denaturation using circular dichroism, fluorescence and UV-difference spectroscopy. Differential effect of buffer composition on GdnHCl denaturation of BLA was evident from the magnitude of these spectral signals, which followed the order: sodium phosphate > Tris-HCl > HEPES > MOPS. These effects became more pronounced with calcium-depleted BLA. Sephacryl S-200 gel chromatographic results showed significant BLA aggregation in the presence of 6 M GdnHCl.
authors.forename: Salanee
authors.initials: S
authors.lastname: Kandandapani
authors.forename: Cheau Y
authors.initials: CY
authors.lastname: Tan
authors.forename: Adawiyah S
authors.initials: AS
authors.lastname: Shuib
authors.affiliation: Biomolecular Research Group, Biochemistry Programme, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia. saadtayyab2004@yahoo.com.
authors.forename: Saad
authors.initials: S
authors.lastname: Tayyab
mesh_terms: D000069976:Bacillus licheniformis / Q000201:enzymology*; D002021:Buffers; D002122:Calcium Chloride / Q000737:chemistry*; D002942:Circular Dichroism; D019791:Guanidine / Q000737:chemistry*; D006531:HEPES / Q000737:chemistry*; D009025:Morpholines / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry*; D010720:Phosphines / Q000737:chemistry*; D011489:Protein Denaturation; D000516:alpha-Amylases / Q000737:chemistry*
chemical_list: D002021:Buffers; D009025:Morpholines; D010710:Phosphates; D010720:Phosphines; C008550:morpholinopropane sulfonic acid; D000516:alpha-Amylases; C501919:alpha-amylase, Bacillus licheniformis; D019791:Guanidine; D002122:Calcium Chloride; D006531:HEPES; C018279:sodium phosphate; C505069:tris(hydroxymethyl)phosphine