Intrinsic halotolerance of the psychrophilic alpha-amylase from Pseudoalteromonas haloplanktis.
The halotolerance of a cold adapted alpha-amylase from the psychrophilic bacterium Pseudoalteromonas haloplanktis (AHA) was investigated. AHA exhibited hydrolytic activity over a broad range of NaCl concentrations (0.01-4.5 M). AHA showed 28% increased activity in 0.5-2.0 M NaCl compared to that in 0.01 M NaCl. In contrast, the corresponding mesophilic (Bacillus amyloliquefaciens) and thermostable (B. licheniformis) alpha-amylases showed a 39 and 46% decrease in activity respectively. Even at 4.5 M NaCl, 80% of the initial activity was detected for AHA, whereas the mesophilic and thermostable enzymes were inactive. Besides an unaltered fluorescence emission and secondary structure, a 10 degrees C positive shift in the temperature optimum, a stabilization factor of >5 for thermal inactivation and a Delta T(m) of 8.3 degrees C for the secondary structure melting were estimated in 2.7 M NaCl. The higher activation energy, half-life time and T(m) indicated reduced conformational dynamics and increased rigidity in the presence of higher NaCl concentrations. A comparison with the sequences of other halophilic alpha-amylases revealed that AHA also contains higher proportion of small hydrophobic residues and acidic residues resulting in a higher negative surface potential. Thus, with some compromise in cold activity, psychrophilic adaptation has also manifested halotolerance to AHA that is comparable to the halophilic enzymes.
Srimathi
Soundararajan
S
Department of Microbiology, Tuberculosis Research Centre, Mayor Ramanathan Road, Chetput, Chennai, 600 031, India. srimathi_matoria@yahoo.com
Jayaraman
Gurunathan
G
Feller
Georges
G
Danielsson
Bengt
B
Narayanan
Paranji R
PR
D000222:Adaptation, Physiological; D002384:Catalysis; D002942:Circular Dichroism; D006219:Halogens / Q000737:chemistry*; D006868:Hydrolysis; D007700:Kinetics; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D031161:Pseudoalteromonas / Q000201:enzymology / Q000502:physiology*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D000516:alpha-Amylases / Q000378:metabolism*
D006219:Halogens; D000516:alpha-Amylases