Abstract
A new α-amylase was extracted from a recently found strain of Bacillus sp. and purified by ion-exchange chromatography. Sodium dodecyl sulfate polyacrylamide gel electrophoresis showed a single band for the purified enzyme with an apparent molecular weight of 59 kDa. The optimum temperature and pH range of the enzyme were 40–60°C and 4.5–7.5, respectively, and its activation energy was 1.974 kcal/mol. The K m value for the enzyme activity on solubie starch was 4 mg/mL, and the T m values obtained from the circular dichroism (CD) results of thermal unfolding were 78.7 and 80.2°C in the absence and presence of the calcium, respectively. The enzyme was almost completely inhibited by the addition of Fe3+, Mn2+, and Zn2+ and was activated by EDTA, Cr3+, and Al3+. Moreover, it was partially inhibited by Ca2+, Ba2+, Ni2+, and Co2+. Proteolytic digestion of the enzyme using trypsin combined with results from T m using CD and irreversible thermoinactivation suggests that this enzyme can be considered a moderate thermophile with both mild flexibility and rigidity.
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Sajedi, R.H., Naderi-Manesh, H., Khajeh, K. et al. Purification, characterization, and structural investigation of a new moderately thermophilic and partially calcium-independent extracellular α-amylase from Bacillus sp. TM1. Appl Biochem Biotechnol 119, 41–50 (2004). https://doi.org/10.1385/ABAB:119:1:41
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DOI: https://doi.org/10.1385/ABAB:119:1:41