Purification and Partial Characterization of α-Amylase Produced by a Thermo-Halophilic Bacterium Isolate PLS 75

Teuku M. Iqbalsyah, Farah Fajarna, Febriani Febriani

Abstract


Bio-based industries require stable enzymes in a broad range of environmental conditions. Extremophiles have attracted more interests as the source of such enzymes, one of which is α-amylase. This study aimed to purify and characterize α-amylase produced by a thermo-halophilic bacterium PLS 75 isolated from under water fumaroles. Ammonium sulfate precipitation results showed that the highest specific α-amylase activity (21.7 U/mg) obtained at 40-60% saturation level, with a purity of 7.7-fold of the crude extract with 16.2% yield. Further purification using DEAE Sepharose column chromatography increased the enzyme purity 11.1-fold of the crude extract with 7.1%. Specific activity after column chromatography purification was 31.3 U/mg. The pure enzyme had a low molecular weight of 14 kDa. The enzyme showed the highest activity at 80 °C and pH 5. The activity increased to 126% when in methanol, while decreased when in ethyl acetate and chloroform. The characteristics of α-amylase with low molecular weight, which was active in acidic condition, stable in polar and non-polar solvents, may be used for for specific industrial needs.


Keywords


α-amylase; DEAE Sepharose; Halostabile; PLS; Thermostabile



DOI: https://doi.org/10.15294/biosaintifika.v10i3.15861

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Biosaintifika: Journal of Biology & Biology Education by Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Negeri Semarang is licensed under a Creative Commons Attribution 4.0 International Licensep-ISSN (Print) 2085-191X | e-ISSN (Online) 2338-7610