ISSN 2456-0235

International Journal of Modern Science and Technology

INDEXED IN 

​​​​​​International Journal of Modern Science and Technology, Vol. 2, No. 3, 2017, Pages 90-97. 

​

​Ameliorating the phytate-degrading phytase production using an isolated Bacillus sp. from agricultural fields

M. Seenuvasan¹,*, J. Iyyappan², M. Anil Kumar³, K. Vaithianathan³
​¹Department of Petrochemical Engineering, SVS College of Engineering, Coimbatore. India.
²Department of Biotechnology, Vel Tech High Tech Dr. Rangarajan Dr. Sakunthala Engineering College, Chennai. India.
³Department of Biotechnology, Madha Engineering College, Chennai. India.
*Corresponding author’s e-mail: msvasan.chem@gmail.com

​Abstract
Accumulation of Phosphate in environment can be reduced by activity of Phytase enzyme molecules. Phytases are the enzymes that instigate removal of phosphate form Phytic acid. An indigenous Bacillus sp. was screened from the vicinities of agricultural fields for the production of phytase enzyme when grown in phytase specific media. The enzyme was secreted extracellularly and the isolate exhibited high activities. The production media was optimized using statistical techniques and the quantified operational parameters were used to perform experiments in bench-top batch fermenter. The phytase was recovered and purified using ammonium sulphate precipitation, dialysis, and chromatographic techniques. The purified enzyme was characterized for its operational stability under varying pH, temperature, inducers. The purified phytase enzyme was optimally active at pH 5, 30°C and enzyme activity was stimulated by CuCl2. The kinetic parameters were evaluated and purified enzyme had specific activity and molecular weight of 3.49 U/mL and 45 kDa, respectively. 

​​Keywords: Phytase; Bacillus species; Optimization; Purification; Characterization; Molecular weight.

​References

  1. Maurya AK, Parashar D, Satyanarayana T. Bioprocess for the production of recombinant HAP phytase of the thermophilic mold Sporotrichum thermophile and its structural and biochemical characteristics. International Journal of Biological Macromolecules. 2017;94:36-44.
  2. Salome KR, Beazley MJ, Webb SM, Sobecky PA, Taillefert N. Biomineralization of U(VI) phosphate promoted by microbially-mediated phytate hydrolysis in contaminated soils. Geochimica et Cosmochimica Acta. 2017; 197:27-42.
  3. Barrientos L, Scott JJ, Murthy PP. Specificity of hydrolysis of phytic acid by alkaline phytase from lily pollen. Plant Physiol. 1994;106:1489-1495.
  4. Cromwell GL. Antimicrobial and promicrobial agents. In: Lewis AJ, Southern LL (Editors). Swine nutrition. 2nd edition. Boca Raton FL: CRC Press;2001;401-421.
  5. McCormick K, Walk CL, Wyatt CL, Adeola O. Phosphorus utilization response of pigs and broiler chickens to diets supplemented with antimicrobials and phytase. Animal Nutrition, 2017; 3(1):77-84.
  6. Rezaei M, Borbor S, Zaghari M, and Teimouri A. Effect of phytase supplementation on nutrients availability and performance of broiler chicks. International Journal of Poultry Science. 2007;6(1):55-58.
  7. Remus J. Poultry and environment reap the benefits of new generation phytase. Feedtech: International Feed Production and Applied Nutrition. 2005;9:22-25.
  8. Megeed A and Tahir A. Reduction of phosphorus pollution from broilers waste through supplementation of wheat based broilers feed with phytase. Journal of Chemistry. 2015:67:1-3.
  9. Gujar PD, Bhavsar KP, Khire JM. Effect of phytase from Aspergillus niger on plant growth and mineral assimilation in wheat (Triticum aestivum Linn.) and its potential for use as a soil amendment. J Sci Food Agric. 2013;93 (9):2242-2247.
  10. Mittal A, Singh G, Goyal V, Yadav A. and Kumar N. Optimization of medium components for phytase production on orange peel flour by Klebsiella sp. DB3 using response surface methodology. Innovative Romanian Food Biotechnology. 2011;9:35-44.
  11. Vohra A, Satyanarayana T. Phytase production by the yeast, Pichia anomala. Biotechnol Lett. 2001;23(7):551-554.
  12. Chanderman A, Puri AK, Permaul K, Singh S. Production, characteristics and applications of phytase from a rhizosphere isolated Enterobacter sp. ACSS,  Bioprocess Biosyst Eng. 2016;39(10):1577-1587.
  13. Sneath PHA. Endospore-forming Grampositive rods and cocci. In: Bergey’s Manual of Systematic Bacteriology Williams & Wilkins., Baltimore, MD. 1986.
  14. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Bio Chem. 1951;193:264-275.
  15. Heinonen JK, Lahti RJ. Analytical Biochemistry. 1981;113:313-317.
  16. Vohra A, Satyanarayana T. Statistical optimization of the medium components by response surface methodology to enhance phytase production by Pichia anomala. Process Biochemistry. 2002;37:999-1004.
  17. Porto TS, Monteiro TIR, Moreira KA, Lima-Filho JL, Silva MPC, Porto ALF, Carneiro-da-Cunha MG. Liquid–liquid extraction of an extracellular alkaline protease from fermentation broth using aqueous two-phase and reversed micelles systems. World J Microbiol Biotechnol. 2005;21:655-659.
  18. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680-686.
  19. Blum H, Beier H, Gross HJ. Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis. 1987;8:93-99.
  20. Choi YM, Suh HJ, Kim JM. Purification and Properties of Extracellular Phytase from Bacillus sp. KHU-10. Journal of Protein Chemistry. 2001;20(4):287-292​.

ISSN 2456-0235