Production of alpha-amylase from Aspergillus oryzae for several industrial applications in a single step

PORFIRI, MA. CECIIA; MILATICH, ESTEBAN J; FARRUGGIA, BEATRIZ; ROMANINI, DIANA

JOURNAL OF CHROMATOGRAPHY B

ELSEVIER

Lugar: Amsterdam; Año: 2016 vol. 1022 p. 87 - 92

0378-4347

A one-step method as a strategy of alpha-amylase concentration and purification was developed in this work. This methodology requires the use of a very low concentration of biodegradable polyelectrolyte (Eudragit® E-PO) and represents a low cost, fast, easy to scale up and non-polluting technology. Besides,this methodology allows recycling the polymer after precipitation.The formation of reversible soluble/insoluble complexes between alpha-amylase and the polymer Eudragit® E-PO was studied, and their precipitation in selected conditions was applied with bioseparation purposes. Turbidimetric assays allowed to determine the pH range where the complexes are insoluble (4.50-7.00); pH 5.50 yielded the highest turbidity of the system. The presence of NaCl (0.05 M) in the medium totally dissociates the protein-polymer complexes.When the adequate concentration of polymer was added under these conditions to a liquid culture of Aspergillus oryzae, purification factors of alpha-amylase up to 7.43 and recoveries of 88% were obtained in a simple step without previous clarification. These results demonstrate that this methodology is suitable for the concentration and production of alpha-amylase from this source and could be applied at the beginning of downstream processing