Premium
Protein‐coated polymer as a matrix for enzyme immobilization: Immobilization of trypsin on bovine serum albumin‐coated allyl glycidyl ether–ethylene glycol dimethacrylate copolymer
Author(s) -
Jasti Lakshmi Swarnalatha,
Dola Sandhya Rani,
Kumaraguru Thenkrishnan,
Bajja Sreedhar,
Fadnavis Nitin W.,
Addepally Uma,
Rajdeo Kishor,
Ponrathnam Surendra,
Deokar Sarika
Publication year - 2014
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.1871
Subject(s) - bovine serum albumin , immobilized enzyme , glutaraldehyde , chemistry , ethylene glycol , polymer , covalent bond , ethylene glycol dimethacrylate , copolymer , polymer chemistry , adsorption , trypsin , chromatography , organic chemistry , enzyme , methacrylic acid
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). © 2014 American Institute of Chemical Engineers Biotechnol. Prog ., 30:317–323, 2014