title: Electrospun polyvinyl alcohol/bovine serum albumin biocomposite membranes for horseradish peroxidase immobilization.
abstract: Electrospinning, a simple and versatile method to fabricate nanofibrous supports, has attracted attention in the field of enzyme immobilization. Biocomposite nanofibers were fabricated from mixed PVA/BSA solution and the effects of glutaraldehyde treatment, initial BSA concentration and PVA concentration on protein loading were investigated. Glutaraldehyde cross-linking significantly decreased protein release from nanofibers and BSA loading reached as high as 27.3% (w/w). In comparison with the HRP immobilized into the nascent nanofibrous membrane, a significant increase was observed in the activity retention of the enzyme immobilized into the PVA/BSA biocomposite nanofibers. The immobilized HRP was able to tolerate much higher concentrations of hydrogen peroxide than the free enzyme and thus the immobilized enzyme did not demonstrate substrate inhibition. The immobilized HRP retained∼50% of the free enzyme activity at 6.4mM hydrogen peroxide and no significant variation was observed in the KM value of the enzyme for hydrogen peroxide after immobilization. In addition, reusability tests showed that the residual activity of the immobilized HRP were 73% after 11 reuse cycles. Together, these results demonstrate efficient immobilization of HRP into electrospun PVA/BSA biocomposite nanofibers and provide a promising immobilization strategy for biotechnological applications.
authors.affiliation: Department of Biotechnology, College of Science, University of Tehran, Tehran, Iran.
authors.forename: Ramin
authors.initials: R
authors.lastname: Fazel
authors.affiliation: Department of Biotechnology, College of Science, University of Tehran, Tehran, Iran.
authors.forename: Seyed-Fakhreddin
authors.initials: SF
authors.lastname: Torabi
authors.affiliation: Department of Biotechnology, College of Science, University of Tehran, Tehran, Iran.
authors.forename: Pooya
authors.initials: P
authors.lastname: Naseri-Nosar
authors.affiliation: Department of Biotechnology, College of Science, University of Tehran, Tehran, Iran.
authors.forename: Salehe
authors.initials: S
authors.lastname: Ghasempur
authors.affiliation: Nanobiotechnology Engineering Laboratory, Faculty of Engineering and New Technologies, Shahid Beheshti University, GC, Tehran, Iran; Protein Engineering Laboratory, Protein Research Center (PRC), Shahid Beheshti University, GC, Tehran, Iran. Electronic address: O_Ranaei@sbu.ac.ir.
authors.forename: Seyed-Omid
authors.initials: SO
authors.lastname: Ranaei-Siadat
authors.affiliation: Department of Biotechnology, College of Science, University of Tehran, Tehran, Iran; Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran. Electronic address: khajeh@modares.ac.ir.
authors.forename: Khosro
authors.initials: K
authors.lastname: Khajeh
mesh_terms: D000818:Animals; D001709:Biotechnology; D002417:Cattle; D003432:Cross-Linking Reagents; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005976:Glutaral; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D008567:Membranes, Artificial; D008855:Microscopy, Electron, Scanning; D008958:Models, Molecular; D053761:Nanocomposites / Q000737:chemistry / Q000648:ultrastructure; D057139:Nanofibers / Q000737:chemistry / Q000648:ultrastructure; D036103:Nanotechnology; D011142:Polyvinyl Alcohol; D012710:Serum Albumin, Bovine / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D008567:Membranes, Artificial; D012710:Serum Albumin, Bovine; D011142:Polyvinyl Alcohol; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D005976:Glutaral
keywords: Biocomposite nanofibrous membranes; Bovine serum albumin; Cross-linking; Electrospinning; Horseradish peroxidase; Immobilization