Protein-coated polymer as a matrix for enzyme immobilization: immobilization of trypsin on bovine serum albumin-coated allyl glycidyl ether-ethylene glycol dimethacrylate copolymer.
Allyl glycidyl ether (AGE)-ethylene glycol dimethacrylate (EGDM) copolymer with 25% crosslink density (AGE-25) shows excellent bovine serum albumin (BSA) adsorption (up to 16% (w/w)) at pH 8.0 and the adsorbed BSA is strongly bound. This protein-coated polymer provides a novel matrix with naturally existing functional groups such as thiol, amino, and carboxylic acid that are available for covalent immobilization of functional enzymes. Employing appropriate strategies, trypsin as a model protein was covalently bound to BSA-coated matrix both independently, and in a stepwise manner on the same matrix, with less than 5% loss of enzyme activity during immobilization. Glutaraldehyde crosslinking after immobilization provide stable enzyme preparation with activity of 510 units/g recycled up to six times without loss of enzyme activity. AFM studies reveal that the polymer surface has protein peaks and valleys rather than a uniform monolayer distribution of the protein and the immobilized enzyme preparation can best be described as polymer supported cross-linked enzyme aggregates (CLEAs).
Indian Inst. of Chemical Technology, Uppal Road, Hyderabad, India.
Lakshmi Swarnalatha
LS
Jasti
Sandhya Rani
SR
Dola
Thenkrishnan
T
Kumaraguru
Sreedhar
S
Bajja
Nitin W
NW
Fadnavis
Uma
U
Addepally
Kishor
K
Rajdeo
Surendra
S
Ponrathnam
Sarika
S
Deokar
D000818:Animals; D002417:Cattle; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D004852:Epoxy Compounds / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008689:Methacrylates / Q000737:chemistry*; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000378:metabolism; D013696:Temperature; D014357:Trypsin / Q000737:chemistry* / Q000378:metabolism
D004800:Enzymes, Immobilized; D004852:Epoxy Compounds; D008689:Methacrylates; D012710:Serum Albumin, Bovine; C004919:ethylene dimethacrylate; D014357:Trypsin; C054361:allyl glycidyl ether
allyl glycidyl ether; bovine serum albumin; ethylene glycol dimethacrylate; immobilization; trypsin