Purification, Characterization and Application of Polygalacturonase from Aspergillus niger CSTRF

Arotupin, D. J., Akinyosoye, F. A., Onifade, A. K.
2012 Malaysian Journal of Microbiology  
Aims: The research was carried out to study the purification, characterization and application of polygalacturonase from Aspergillus niger CSTRF. Methodology and Results: The polygalacturonase (PG) from the fungus was purified by ammonium sulphate precipitation and dialysed. The resulting fraction of the enzyme was further separated by molecular exclusion and ion exchange chromatography. The enzyme was purified 28.19 fold with a yield of approximately 69 % following purification with SP C-50.
more » ... has a relative molecular weight of 79,430 daltons and markedly influenced by temperature, pH and substrate concentrations of reactions with optimum activity at 35 °C, pH 4.0 and 8 mg/mL respectively. The PG was heat stable over a broad range of temperatures. Line weaver-Burk plot for the apparent hydrolysis of pectin showed approximately Km value of 2.7 mg/mL. The activity of the enzyme was enhanced by Na + , Ca 2+ , Mg 2+ and Zn 2+ , while EDTA, PbCl2, HgCl2 and IAA were inhibitory. The ability of the purified enzyme to clarify fruit juice was also investigated. Conclusion, significance and impact of the study: This study revealed that polygalacturonase possesses properties for clarification of fruit juice and by extension bioprocessing applications. ISSN (print): 1823-8262, ISSN (online): 2231-7538 2621. Caprari and C.; Bergmann, C.; Migheli, Q.; Salvi G.;Alberbersheim, P. Darvill, A.; Cervone, F. and De Lorenzo, G. (1993). Fusarium moniliforme secrectes four endopolygalacturonases derived from a single gene product. Physiological and Molecular Plant Pathology 43 : 453 -462. Casida, L. E. Jr. (1997). Industrial Microbiology. Enzymes as Fermentation Products 4 th edition, Wiley Eastern Limited New Delhi pp.390 -401. Chuku, E. C.; Onuegbu, B. A. and Osakwe, J. A. (2005). Activities of polygalacturonase and cellulose enzymes on Viscosity of the seed of Irvingia gabonesis (Bush Mango). Niger Delta Biologia 5(1): 76-78. Conn, E. E. and Stump K. P. (1989). Outline of Biochemistry, 4 th edition. Wiley Eastern Limited, New Delhi, India pp. 629 Creighton, T. E. (1990). Protein Function: A practical Approach. Oxford University Press, Oxford 306pp. Crotti, L. B.; Jabopr, V. A.; Chellegatti, M. A.; fonseca, M. J. and Said, S. (1999). Studies of pectic enzymes produced by Talaromyces flavus in submerged and solid substrate cultures. Journal of Basic Microbiology, 39: 49-260. Daniel, R. M.; Dines, M. and Petach, H. H. (1996). The denaturation and degradation of stable enzymes at high temperatures. Biochemical Journal 317: 1 -11. Debing, J.; Peijun, L.; Stagnitti, F. Xianzhe, X. and Li, L. (2006). Pectinase production by soild fermentation from Aspergillus niger by a new prescription experiment. Ecotoxicology and Environmental Safety 64:244-250. Dixon, M. and Webbs, E. C. (1971). Enzymes Williams Clowes and Sons. Great Britain 950 337pp. Dudchenko, L. G. (1986). Effect of metal ions on the activity of pectolytic enzymes of the higher basidia fungus Coriolus hirsutuo. Geol Khim Biol Nauk 10:59 -62. Famurewa. O. Oyede. M. A. and Olutiola P. O. (1993). Pectin transeliminase complex in culture filterates of Aspergillus flavus. Folia Microbiol. 38: 459 -466. Favela-Torres, E.; Volke-Sepulveda, T. and Viniegra-Gonzalez, G. (2006). Production of hydrolytic depolymerising pectinases. Food Technology and Biotechnology 44(2):221-227. Fontana, R.C.; Salvador, S. and Da Silveira M.M. (2005). Influence of pectin and glucose on growth and polygalacturonase production by Aspergillus niger in solid-state cultivation.
doi:10.21161/mjm.04512 fatcat:3mtqlngvc5gmvggzrkvvxbseqm