Recovery of pectinases from Aspergillus niger using aqueous two-phase systems / Recuperação de pectinases de Aspergillus niger usando sistemas aquosos bifásicos

Márcia M. Santin Trentini, Anne Luize Lupatini Menegotto, Juliana Steffens, Jamile Zeni, Geciane Toniazzo Backes, Rogério M. Dallago, Marco Di Luccio, Eunice Valduga

Abstract


This work aimed to study the purification of pectinases by aqueous two-phase systems (ATPS). The crude enzymatic extract was produced by Aspergillus niger ATCC 9642 and contained exo-polygalacturonase (exo-PG), pectinmethylesterase (PME) and pectin lyase (PMGL). The ATPS systems tested consisted in the combinations of polyethylene glycol (PEG) and potassium phosphate and sodium citrate, alcohol (ethanol, n-propanol and isopropanol) and salt (ammonium sulfate, potassium phosphate and sodium citrate). The experiments showed higher recoveries using ATPS system - PEG/phosphate for the exo-PG were using 16% PEG 4.0 kDa/4.8% NaCl and 16% PEG 1.5 kDa/without NaCl, obtaining purification factors (PF) of 1.37 and 1.21 times and recovery (R) of 49 and 59%, respectively. However, for the enzymes PME and PMGL were of 4.8 and 4.7 fold and 478 and 241%, respectively. When used ATPS system - PEG/sodium citrate the best PF were of 2.4, 7.85 and 5.7 and R of 100, 331 and 239% for exo-PG, PME and PMGL, respectively. The ATPS system is an alternative and efficient method for the recovery and/or purification of pectinases.

 


Keywords


alcohol, enzymes, polyethylene glycol, purification, salt.

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References


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DOI: https://doi.org/10.34117/bjdv6n7-427

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