Protein‐coated polymer as a matrix for enzyme immobilization: Immobilization of trypsin on bovine serum albumin‐coated allyl glycidyl ether–ethylene glycol dimethacrylate copolymer |
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| Authors: | Lakshmi Swarnalatha Jasti Sandhya Rani Dola Thenkrishnan Kumaraguru Sreedhar Bajja Nitin W. Fadnavis Uma Addepally Kishor Rajdeo Surendra Ponrathnam Sarika Deokar |
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| Affiliation: | 1. Indian Inst. of Chemical Technology, Hyderabad, India;2. Centre for Biotechnology, Jawaharlal Nehru Technological University, Hyderabad, India;3. Polymer Science & Engineering, Chemical Engineering Division, National Chemical Laboratory, Pune, India |
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| Abstract: | 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 |
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| Keywords: | trypsin bovine serum albumin immobilization allyl glycidyl ether ethylene glycol dimethacrylate |
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