Isolation and Activity Determination of Enzyme Phosphatase Secreted by Aspergillus niger

  • Tafadzwa Zharare Department of Biochemistry, University of Zimbabwe
  • Rumbidzai Mangoyi Department of Biochemistry, University of Zimbabwe http://orcid.org/0000-0002-0663-3058

Abstract

The use of enzymes on industrial scale saves a lot of energy and avoids pollution, thus holding a promise for green and economically sustainable alternative strategies in industrial transformations.  Generally, the fungi Aspergillus niger secretes enzymes which can be used in different industries. Thus, coming up with these enzymes in large amounts will definitely result in reduced costs encountered in importing them for industrial use.  This study focussed on isolation and activity determination of an enzyme phosphatase secreted by Aspergillus niger.  This enzyme can be of great importance in molecular biology industries, particularly for recombinant DNA technology.  For this study, pure cultures of Aspergillus niger were used.  Aspergillus niger was resuscitated on potato dextrose agar and then subcultured in Adam’s medium, a medium specific for the production of phosphatase.  Cells were centrifuged and the filtrate was collected whilst the residue was discarded. The filtrate was expected to contain the crude enzyme phosphatase since Aspergillus niger secretes the extracellular enzyme into the medium. Disodium phenyl phosphate was used as a substrate for the determination of the phosphatase activity. The enzyme activity was determined spectrophotometrically by reading absorbance of phenol formed in the presence of enzyme and the substrate. The concentration of phenol liberated was then used to calculate the enzyme activity expressed in King Armstrong Units (KAU).  Further work on enzyme activity determination was done by varying enzyme and substrate concentrations.  Results showed that the isolated alkaline phosphatase had activity of 4.0 KAU and 4.5 KAU at 25 ºC and 37 ºC respectively. Acidic phosphatase had activity of 5 KAU and 7 KAU at 25 ºC and 37 ºC respectively. Rate of activity increased upon increasing enzyme concentration and substrate.  Thus, Aspergillus niger produces the enzyme phosphatase, however, there is need to induce the production of these enzymes for industrial use.

Keywords: Activity, acidic phosphatase, alkaline phosphatase, Aspergillus niger, enzymes, industrial use

Downloads

Download data is not yet available.

References


  1. K.M. Engqvist and K.S. Rabe, “Applications of Protein Engineering and Directed Evolution in Plant Research” Plant Physiol, 179, 3,  907-917, March, 2019.

  2. Sindhu, P. Binod, B.U. Sabeela, A. Amith, K.M. Anil, M. Aravind, R. Sharrel and P. Ashok,  “Applications of Microbial Enzymes in Food Industry” Food Technol. Biotechnol, 56, 1, 16-30,  January 2018.

  3. L. Brautigan and S. Shenolikar, “Protein Serine/Threonine Phosphatases: Keys to Unlocking Regulators and Substrates” Annu Rev Biochem, 87, 921-964, June 2018.

  4. Prajapatia, K.D.S.N. Patil and S.V. Chaudhari, “Alkaline Phosphatase: Significance In Dairy Industry” Inter J Res Sc Manage, 4, 12, 18-22, December, 2017.

  5. Ezawa and K. Saito, “How do Arbuscular mycorrhizal fungi handle phosphate? New insight into fine‐tuning of phosphate metabolism” New Phytol, 220, 4, 1116-1121, April, 2018.

  6. Iyer, M.S. RajputS. Rajkumar, “ Effect of succinate on phosphate solubilization in nitrogen fixing bacteria harbouring chick pea and their effect on plant growth” Microb Res, 202, 43-50, September 2017.

  7. Koncki, D. Ogończyk and S. Głąb, (2005).” Potentiometric assay for acid and alkaline phosphatase” Analytica chimica acta, 538, 1, 257-261. April, 2005.

  8. Robinson, “Enzymes: principles and biotechnological applications” Essays Biochem, 59, 1-41, November, 2015.

  9. C. Behera, H. Yadav, S.K. Singh, R.R. Mishra, B.K. Sethi, S.K. Dutta, H.N. Thatoi, “Phosphate solubilization and acid phosphatase activity of Serratia sp. isolated from mangrove soil of Mahanadi river delta, Odisha, India” Jour Genetic Engin Biotech, 1-10, January, 2017.

  10. C. Behera, H. Yadav, S.K. Singh, R.R. Mishra, B.K. Sethi, S.K. Dutta, H.N. Thatoi, “Alkaline phosphatase activity of a phosphate solubilizing Alcaligenes faecalis, isolated from Mangrove soil” Biotech Res Inno, 1, 1, 101-111, December 2017.

  11. L. Purich, “Enzyme kinetics: catalysis & control: a reference of theory and best-practice methods” Elsevier, 1-4, 2010.

Published
2019-11-16
How to Cite
[1]
T. Zharare and R. Mangoyi, “Isolation and Activity Determination of Enzyme Phosphatase Secreted by Aspergillus niger”, Int. Ann. Sci., vol. 9, no. 1, pp. 41-45, Nov. 2019.
Section
Short Communication

Funding data