Pectinases partitioning in aqueous two-phase systems: An integration of the systems poly(ethylene glycol)crude dextran and poly(ethylene glycol)ammonium sulphate

• Published in: Journal of the Serbian Chemical Society 2004-01, Vol.69 (4), p.299-307
• Main Authors: Antov, Mirjana, Pericin, Draginja, Pejin, Stana
• Format: Article
• Language: English
• Subjects: aqueous two-phase system; partitioning; pectinases
• ISSN: 0352-5139; 1820-7421
• DOI: 10.2298/JSC0404299A

The partitioning of pectinases in the poly(ethylene glucol)4000/ammonium sulpohate system was studied and also its application for enzymes extraction from the top phase of the poly(ethylene glucol)4000/crude dextran system. Almost complete one-sided partition of endo-pectinase and exo-pectinase to the bottom phase of the polymer/salt system was achieved at a tie-line length of 37.16 %. The concentration factors were 1.73 and 3.25, respectively. The highest total endo- and exo-pectinase yields (72.41 % and 69.46 % respectively) were obtained by integration of the polymer/polymer system at a tie-line of 8.61 % and a high phase volume ratio and the polymer/salt system at a tie-line of 30.23%and a low phase volume ratio. Integration of the partitioning at a high tie-line length in the polymer/polimer and a low tie-line length in the polymer/salt system resulted in a total concentration factor of 1.5 and a purification of 1.66 fold for exo-pectinase. The addition of phosphate to this integrated system improved the total concentration factor and purification fold of the activity to 1.73 and 2.14, respectively. Ispitivana je raspodela pektinaza u sistemu polietilen glikol 4000/amonijum sulfat, kao i primena u ekstrakciji enzima iz gornje faze sistema polietilen glikol 4000/sirovi dekstran. Dobijena je gotovo potpuna raspodela endo-pektinaze i egzo-pektinaze u donju fazu sistema polimer/so pri duzini vezne linije od 37,16 %, uz faktore koncentrisanja 1,73 i 3,25, redom. Najveci ukupni prinosi endo- i egzo-pektinaze dobijeni su integracijom sistema polimer/polimer, pri veznoj liniji 30,23 % i niskom zapreminskom odnosu faze i iznosili su 72,41%i 69,46 %, redom. Integracijom raspodela na velikoj duzini vezne linije u sistemu polimer/polimer i maloj duzini vezne linije u sistemu polimer/so dobijen je ukupni faktor koncentrisanja 1,5 i stepen pre ciscenosti 1,66 za egzo-pektinazu, a dodatak fosfata u ovako integrisani sistem povecao je vrednosti ovih parametara na 1,73 i 2,14, redom.