@article { ISI:000326660900038, title = {Carrier free co-immobilization of alpha amylase, glucoamylase and pullulanase as combined cross-linked enzyme aggregates (combi-CLEAs): a tri-enzyme biocatalyst with one pot starch hydrolytic activity}, journal = {Bioresource Technology}, volume = {147}, year = {2013}, month = {NOV}, pages = {269-275}, publisher = {ELSEVIER SCI LTD}, address = {THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND}, abstract = {

A tri-enzyme biocatalyst {\textquoteleft}{\textquoteleft}combi-CLEAs{\textquoteright}{\textquoteright} with starch hydrolytic activity was prepared from commercially available alpha amylase, glucoamylase and pullulanase preparations by aggregating enzymes with ammonium sulphate followed by cross-linking formed aggregates for 4.5 h with 40 mM glutaraldehyde. The effects of precipitant type and cross-linking were studied and the biocatalyst was characterized. Scanning electron microscopy analysis showed that tri-enzyme biocatalyst was of spherical structure. For one pot starch hydrolytic activity, shift in optimum pH from 6 to 7 and temperature from 65 to 75 degrees C were observed after co-immobilization of enzymes. After one pot starch hydrolysis reaction in batch mode, 100\%, 60\% and 40\% conversions were obtained with combi-CLEAs, separate CLEAs mixture and free enzyme mixture, respectively. Co-immobilization also enhanced the thermal stability of enzymes. Finally, the catalytic activity of enzymes in combi-CLEAs during one pot starch hydrolysis was well maintained up to five cycles without performance changes. (C) 2013 Elsevier Ltd. All rights reserved.

}, keywords = {Alpha amylase, Combined cross-linked enzyme aggregates (combi-CLEAs), Glucoamylase, One pot starch hydrolysis, Pullulanase}, issn = {0960-8524}, doi = {10.1016/j.biortech.2013.08.035}, author = {Talekar, Sachin and Pandharbale, Amol and Ladole, Mayur and Nadar, Shamraja and Mulla, Mosin and Japhalekar, Kshitija and Pattankude, Kishori and Arage, Devika} }