Immobilization of polygalacturonase from Aspergillus niger onto activated polyethylene and its application in apple juice clarification.
The present work is focused on efficient immobilization of polygalacturonase on polyethylene matrix, followed by its application in apple juice clarification. Immobilization of polygalacturonase on activated polyethylene and its use in apple juice clarification was not reported so far. Aspergillus niger Van Tieghem (MTCC 3323) produced polygalacturonase when grown in modified Riviere's medium containing pectin as single carbon source by fed-batch culture. The enzyme was precipitated with ethanol and purified by gel filtration chromatography (Sephacryl S-100) and immobilized onto glutaraldehyde-activated polyethylene. The method is very simple and time saving for enzyme immobilization. Various characteristics of immobilized enzyme such as optimum reaction temperature and pH, temperature and pH stability, binding kinetics, efficiency of binding, reusability and metal ion effect on immobilized enzymes were evaluated in comparison to the free enzyme. Both the free and immobilized enzyme showed maximum activity at a temperature of 45 degrees C and pH 4.8. Maximum binding efficiency was 38%. The immobilized enzyme was reusable for 3 cycles with 50% loss of activity after the third cycle. Twenty-four U of immobilized enzyme at 45 degrees C and 1 h incubation time increased the transmittance of the apple juice by about 55% at 650 nm. The immobilized enzyme can be of industrial advantage in terms of sturdiness, availability, inertness, low price, reusability and temperature stability.
Department of Biotechnology, Himachal Pradesh University, Summerhill, Shimla-171 005, India.
Shivalika
S
Saxena
Surendra
S
Shukla
Akhilesh
A
Thakur
Reena
R
Gupta
D001234:Aspergillus niger / Q000201:enzymology* / Q000254:growth & development; D001628:Beverages; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D003067:Coenzymes / Q000494:pharmacology; D003470:Culture Media; D004795:Enzyme Stability; D005511:Food Handling / Q000379:methods*; D005516:Food Microbiology; D006863:Hydrogen-Ion Concentration; D027845:Malus / Q000378:metabolism; D008670:Metals / Q000494:pharmacology; D010368:Pectins / Q000378:metabolism; D020959:Polyethylene; D011096:Polygalacturonase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D011485:Protein Binding; D013696:Temperature
D003067:Coenzymes; D003470:Culture Media; D008670:Metals; D010368:Pectins; D020959:Polyethylene; D011096:Polygalacturonase