title: Graft copolymerization of acrylamide on chitosan-co-chitin and its application for immobilization of Aspergillus sp. RL2Ct cutinase.
abstract: The synthesis of chitosan (Chs) and chitin (Chi) copolymer and grafting of acrylamide (AAm) onto the synthesized copolymer have been carried out by chemical methods. The grafted copolymer was characterized by FTIR, SEM and XRD. The extracellular cutinase of Aspergillus sp. RL2Ct (E.C. 3.1.1.3) was purified to 4.46 fold with 16.1% yield using acetone precipitation and DEAE sepharose ion exchange chromatography. It was immobilized by adsorption on the grafted copolymer. The immobilized enzyme was found to be more stable then the free enzyme and has a good binding efficiency (78.8%) with the grafted copolymer. The kinetic parameters KMand Vmaxfor free and immobilized cutinase were found to be 0.55mM and 1410μmolmin-1mg-1protein, 2.99mM and 996μmolmin-1mg-1protein, respectively. The immobilized cutinase was recycled 64 times without considerable loss of activity. The matrix (Chs-co-Chi-g-poly(AAm)) prepared and cutinase immobilized on the matrix have potential applications in enzyme immobilization and organic synthesis respectively.
authors.affiliation: Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla 171 005, India.
authors.forename: Vijaya
authors.initials: V
authors.lastname: Kumari
authors.affiliation: Department of Chemistry, Himachal Pradesh University, Summer Hill, Shimla 171 005, India.
authors.forename: Sandeep
authors.initials: S
authors.lastname: Kumar
authors.affiliation: Department of Chemistry, Himachal Pradesh University, Summer Hill, Shimla 171 005, India.
authors.forename: Inderjeet
authors.initials: I
authors.lastname: Kaur
authors.affiliation: Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla 171 005, India. Electronic address: bhallatc@rediffmail.com.
authors.forename: Tek Chand
authors.initials: TC
authors.lastname: Bhalla
mesh_terms: D020106:Acrylamide / Q000138:chemical synthesis / Q000737:chemistry*; D000327:Adsorption; D001230:Aspergillus / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D002686:Chitin / Q000138:chemical synthesis / Q000737:chemistry*; D048271:Chitosan / Q000138:chemical synthesis / Q000737:chemistry*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D058105:Polymerization
chemical_list: D004800:Enzymes, Immobilized; D002686:Chitin; D020106:Acrylamide; D048271:Chitosan; D002265:Carboxylic Ester Hydrolases; C019952:cutinase
keywords: Acrylamide; Aspergillus sp. RL2Ct; Chitin; Chitosan; Copolymer grafting; Cutinase; Immobilization