Purification and Characterization of Contemporaneously Produced Alkaline Protease and α-amylase Enzymes from Locally Isolated Bacillus methylotrophicus SCJ4

Authors

  • Hamada El-Gendi Bioprocess Development Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City for Scientific Research and Technology Applications, Alexandria, Egypt.
  • Mohamed S. Azab Botany and Microbiology Dept., Faculty of Science, Al-Azhar University, Cairo, Egypt.
  • Esmail M. El-Fakharany Protein Research Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City for Scientific Research and Technology Applications, Alexandria, Egypt.
  • Nadia A. Soliman Bioprocess Development Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City for Scientific Research and Technology Applications, Alexandria, Egypt.
  • Yasser R. Abdel-Fattah Bioprocess Development Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City for Scientific Research and Technology Applications, Alexandria, Egypt.

Keywords:

Purification, Characterization, Alkaline Protease, α-amylase, Bacillus methylotrophicus

Abstract

The contemporaneously produced alkaline protease and α-amylase by locally isolated Bacillus methylotrophicus SCJ4 had been purified in two steps and some characteristics of the purified enzymes were studied. Crude enzymes were first salted out using ammonium sulfate, and then concentrated proteins were applied to anion column chromatography Q-sepharose (HiPrep Q FF 16/10 column). The purified alkaline protease has an estimated molecular mass of 24 kDa with maximum enzyme activity (1185U/ml/min) at pH 9.0 and 55ºC. This enzyme belongs to serine proteases family with remarkable stability up to 62% and 61.5% toward Sodium Dodecyl Sulfate (SDS) and Hg++ respectively. On the other hand, the purified amylase is a calcium-independent α-amylase, showing a good stability against SDS up to 73%. The estimated molecular mass was 67.5 kDa with maximum enzyme activity (280U/ml/min) at pH 7.0 and 65ºC.

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Published

2016-09-30

How to Cite

El-Gendi, H., Azab, M. S., El-Fakharany, E. M., Soliman, N. A., & Abdel-Fattah, Y. R. (2016). Purification and Characterization of Contemporaneously Produced Alkaline Protease and α-amylase Enzymes from Locally Isolated Bacillus methylotrophicus SCJ4. PSM Biological Research, 1(2), 88–95. Retrieved from https://psmjournals.org/index.php/biolres/article/view/39

Issue

Vol. 1 No. 2 (2016)

Section

Articles