An immobilized fungal chlorogenate hydrolase for the production of quinic and caffeic acids from yerba mate extract

BUTIUK, A.; ADACHI, O.; MIGNONE, C.; HOURS, R.

Mar del Plata

Encuentro; VIII Encuentro Latinoamericano y del Caribe de Biotecnología; 2013

REDBIO Argentina 2013

Chlorogenate hydrolase (EC 3.1.1.42, CHase) catalyzes chlorogenic acid (CGA) hydrolysis releasing quinic and caffeic acids. It was highly produced in mycelia of Aspergillus niger AKU 3302 grown in Czapek medium containing 0.125% w/v of concentrated yerba mate extract as inducer. Mycelial growth and spore germination were negative after a heat treatment at 55°C for 60 min without any appreciable loss of enzyme activity. The heated mycelium thus prepared was packed in a column and used as immobilized biocatalyst. The column was washed several times with Na-phosphate buffer containing EDTA and then 0.1 M CGA solution was loaded through the CHase-immobilized column under various conditions (different cycles, flow rates and reaction times). No gas bubble formation was observed. The immobilized CHase followed a first order enzyme kinetics and CGA concentration decayed linearly with reaction time. Then, a solution containing 0.6% (w/v) of yerba mate extract was circulated at room temperature through the CHase-immobilized column. The initial CGA in yerba mate solution was almost completely hydrolyzed after 3 h of circulation, because of no further spectral change was observed even though the incubation was prolonged. Under these conditions, CHase-immobilized may be applied to practical production of quinic and caffeic acids.