PURIFICATION AND DETERMINATION OF PHYSICOCHEMICAL PROPERTIES OF α-AMYLASE SECRETED BY PENICILLIUM CITRINUM S1
DOI:
https://doi.org/10.22159/ajpcr.2025v18i12.56990Keywords:
Penicillium citrinum S1, α-amylase, purification, characterization, SDS-PAGE, specific activityAbstract
Objective: The α-amylase is a key industrial enzyme with widespread applications having about 30% share in the world enzyme market. The purpose of the study was to purify the α-amylase produced from Penicillium citrinum S1 and to evaluate the physicochemical and kinetic properties of the enzyme for its potentiality as an industrially valuable enzyme.
Methods: Production of extracellular α-amylase was done by using P. citrinum S1 in liquid medium and subjected to purification of the enzyme through ammonium sulfate precipitation, dialysis, followed by gel filtration chromatography.
Results: The enzyme was purified to 10.05 fold with a specific activity of 23.32 μg/mL. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis of the enzyme revealed that the molecular weight of the enzyme is ≈43 kDa. From enzyme kinetic study, the Km value of α-amylase was found as 1.1 μg/mL, and the Vmax value was 37.31 U/mL. The optimum temperature showing the highest enzyme reaction is 50°C, and the most ideal pH range is 6.5–7.5. The purified enzyme remained stable at moderately high temperature (50–60°C) and up to 120 min and remained active in alkaline medium (0.1%) of commercial detergents such as Rin and Surf Excel.
Conclusion: These distinctive properties of this α-amylase purified from P. citrinum S1, such as low Km value, wide range of pH tolerance, stability of the enzyme at 50–60°C, compatibility with commercial detergents such as rin and surf, may be promising for application in starch processing, brewing, textile desizing, and detergent formulations.
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