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    A Novel Raw Starch Hydrolyzing Thermostable ?-Amylase Produced by Newly Isolated Bacillus mojavensis SO-10: Purification, Characterization and Usage in Starch Industries
    (Inst Tecnologia Parana, 2018) Ozdemir, Sadin; Fincan, Sema Aguloglu; Karakaya, Adem; Enez, Baris
    The aim of this study is the production, purification, and characterisation of thermostable raw starch hydrolyzing aamylase produced by Bacillus mojavensis SO-10. The maximum production conditions of alpha-amylase were found at 36th hour, 35 degrees C and pH 7.0. We utilized three steps to purify the thermostable alpha-amylase and as a result, 34-fold and 18% yield were obtained. The molecular weight of purified alpha-amylase was determined as 73 kD. The K-m and V-max, rates were detected as 0.010 mM and 3.38 ,umol min(-1), respectively. This purified alpha-amylase exhibited the highest activity at pH 5.0-6.0 and 70 degrees C and showed stability over a wide variety of pH and temperature at 4.0-8.0, and 4050 degrees C, respectively. The thermostable purified alpha-amylase exhibited stability in the presence of denaturing agents and heavy metal ions. The purified enzyme hydrolyzed the raw starches of corn and wheat grains in the ratio of 36.7% and 39.2% respectively. The end-yields of soluble starch hydrolysis were analyzed by thin-layer chromatography (TLC). In addition, the usage of purified a-amylase in clarification of apple juice and domestic washing detergent industries were evaluated.
  • [ X ]
    Öğe
    Purification and characterization of thermostable a-amylase produced from Bacillus licheniformis So-B3 and its potential in hydrolyzing raw starch
    (Pergamon-Elsevier Science Ltd, 2021) Fincan, Sema Aguloglu; Ozdemir, Sadin; Karakaya, Adem; Enez, Baris; Mustafov, Sibel Demiroglu; Ulutas, Mehmet Sefa; Sen, Fatih
    Aims: This work was achieved to obtain the optimum culture conditions of the thermostable alpha-amylase produced by thermophilic Bacillus licheniformis SO-B3. Furthermore, the alpha-amylase was purified and then characterized, and also its kinetic parameters were determined. Materials and methods: A new thermotolerant bacteria called Bacillus licheniformis SO-B3 employed in this work was isolated from a sample of thermal spring mud in Sirnak (Meyremderesi). Several parameters such as the impact of temperature, time, and pH on enzyme production were examined. Thin-Layer Chromatography (TLC) was employed to analyze the end-products of soluble starch hydrolysis, and the utilization of purified alpha-amylase in the clarification of unripe apple juices was studied. Key findings: The highest enzyme production conditions were determined as 35 degrees C, 36th hour, and pH 7.0. Thermostable alpha-amylase was purified by 70% ammonium sulfate precipitation, DEAE-cellulose ion-exchange chromatography, and dialysis, with a 51-purification fold and 30% yield recovery. The K-m and V-max values for this enzyme were 0.004 mM and 3.07 mu mol min(-1) at 70 degrees C, respectively. The alpha-amylase's molecular weight was found as 74 kDa. In addition, alpha-amylase showed a good degradation rate for raw starch. Significance: It was hypothesized that Bacillus licheniformis SO-B3 could be used as an alpha-amylase source. These findings displayed that purified enzyme could be utilized in fruit juice industries for clarification of apple juice and raw starch hydrolyzing.