title: A smart bioconjugate of chymotrypsin.
abstract: alpha-Chymotrypsin was immobilized on Eudragit S-100 via covalent coupling with 93% retention of proteolytic activity. The conjugate behaved as a smart biocatalyst and functioned as a pH-dependent reversibly soluble-insoluble biocatalyst. The pH optimum of chymotrypsin broadened on immobilization, and the immobilized preparation showed better stability at and above pH 6.5 as compared to the free enzyme. The immobilized enzyme showed a slight shift in the temperature optimum and enhanced thermal stability retaining 70% of its original activity after 1 h of exposure to 40 degrees C as compared to the 25% residual activity for the free enzyme under identical conditions. K(m) and V(max) values did not change on immobilization. Also, the immobilized preparation was quite stable to reuse, it retained almost 85% of its original activity even after a fifth precipitation cycle. UV spectroscopy and circular dichroism were used to probe structural changes in the enzyme upon immobilization.
authors.affiliation: Chemistry Department, Indian Institute of Technology, Delhi, Hauz Khas, New Delhi 110016, India.
authors.forename: Shweta
authors.initials: S
authors.lastname: Sharma
authors.forename: Prabhjot
authors.initials: P
authors.lastname: Kaur
authors.forename: Aklank
authors.initials: A
authors.lastname: Jain
authors.forename: Moganty R
authors.initials: MR
authors.lastname: Rajeswari
authors.forename: Munishwar N
authors.initials: MN
authors.lastname: Gupta
mesh_terms: D002918:Chymotrypsin / Q000737:chemistry*; D002942:Circular Dichroism; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
chemical_list: D002918:Chymotrypsin; C117608:alpha-chymotrypsin