Fractionation of fungal pectic enzymes by immobilized metal ion affinity chromatography

NAVARRO DEL CAÑIZO, AGUSTÍN; ROQUE HOURS,; MIRANDA, MARÍA VICTORIA; OSVALDO CASCONE,

JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE

Año: 1994 vol. 64 p. 527 - 531

0022-5142

A juice-clarifying fraction that will not produc€ methanol was^obtained loá " -."."."i"f p""tii "-y-" by separating the pectin lyase (EC4 2'2 10) i.á. p""tln"rt"t""" igc¡.r.l.ti). This was achieved by immobilisld m€tal ion utf*ity "ttto-utog.uptry 0tr't¡C) on SepharosFiminodiacetiacc idtu(II) P€ctin lvase áid not bind to the chromatogaphic matrix at pH 8 0' while pectinesterase was ret¿ined and only eluted when the pH of thc buffer was brought down to 3 0. Polvsalacturonasaec rivity( EC3 2.l l5) wasf ound in both fr¿ctionst hus sug- Lri-" iñ"iiprÁc can disciminate between two forms of that c¡zymc Both ii"liio'nt ft"". tbe abilitv to clarify apple juice' thc first without producir¡g methanol. Thc behaviou¡o f eacho f the abovec nzymesin IMAC suggeststh at p€cün iya." tact" acccssibleh istidin€ residues,w hile p€€tinesterasec ould have one or more of them.