Utilization of Integrative Technique for Partial Recovery of Proteases from Soil Microbes

  • Said Nawab Department of Biotechnology, Faculty of Biological Sciences, University of Malakand, Lower Dir, 18800, Khyber Pakhtunkhwa, Pakistan
  • Muhammad Aasim Department of Biotechnology, Faculty of Biological Sciences, University of Malakand, Lower Dir, 18800, Khyber Pakhtunkhwa, Pakistan
  • Haris Saddique Department of Biotechnology, Faculty of Biological Sciences, University of Malakand, Lower Dir, 18800, Khyber Pakhtunkhwa, Pakistan
  • Fazal Rabi Department of Biotechnology, Faculty of Biological Sciences, University of Malakand, Lower Dir, 18800, Khyber Pakhtunkhwa, Pakistan
  • Waqar Ali Department of Biotechnology, Faculty of Biological Sciences, University of Malakand, Lower Dir, 18800, Khyber Pakhtunkhwa, Pakistan
  • Nadir Zaman Khan Department of Biotechnology, Faculty of Biological Sciences, University of Malakand, Lower Dir, 18800, Khyber Pakhtunkhwa, Pakistan

Abstract

Aqueous two-phase system (ATPS) is an efficient, cost effective, fast, simple and ecofriendly method for the recovery of biomolecules. In the present study, an ATPS composed of polyethylene glycol and ammonium sulphate (NH4)2SO4 was used for the partial purification of proteases from microbial source. The effects of different parameters such as molecular weight of PEG (4000, 6000 and 10000), concentration of PEG (15, 17.5 and 20 %) and concentration of (NH4)2SO4 (7.5, 8.3, 9.1 and 9.9 %) on the partitioning behavior of proteases at room temperature were investigated. Generally, increasing the concentration of PEG and (NH4)2SO4 moved the protease to the top i.e., polymer-rich phase. Increasing the molecular weight of PEG from 4000 to 10000 the partition coefficient decreased subsequently. The highest partition coefficient i.e., 3.32 and maximum activity i.e., 16.06 soxhlet unit was found in an optimum system composed of 20 % PEG 4000 and 9.9 % (NH4)2SO4.

Keywords: proteases, aqueous two-phase system, partitioning, partition coefficient, polyethylene glycol (PEG), ammonium sulphate
Published online
2018-11-30
How to Cite
Nawab, S., Aasim, M., Saddique, H., Rabi, F., Ali, W. and Khan, N. “Utilization of Integrative Technique for Partial Recovery of Proteases from Soil Microbes”, Periodica Polytechnica Chemical Engineering. doi: https://doi.org/10.3311/PPch.12637.
Section
Articles