Preparation of nanofibrous polymer grafted magnetic poly(GMA-MMA)-g-MAA beads for immobilization of trypsin via adsorption

被引:84
作者
Bayramoglu, Guelay [1 ,2 ]
Yilmaz, Meltem [1 ]
Senel, Ayseguel Uelkue [2 ]
Arica, M. Yakup [1 ]
机构
[1] Kirikkale Univ, Fac Sci, Biochem Proc & Biomat Res Lab, TR-71450 Kirikkale, Turkey
[2] Kirikkale Univ, Fac Sci, Dept Chem, TR-71450 Kirikkale, Turkey
关键词
magnetic beads; adsorption; enzyme technology; protease; protein; peptide maps;
D O I
10.1016/j.bej.2007.12.013
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Poly (glycidylmethacrylate-methylmethacrylate), poly(GMA-MMA) beads were prepared via suspension polymerization in the presence of ferric ions. The epoxy groups of the poly(GMA-MMA) beads were converted into amino groups during magnetization reaction, and then were grafted with methacrylic acid (MAA) via graft copolymerization. The magnetic beads were characterized by surface area measurement, swelling test, scanning electron microscope (SEM), electron spin resonance (ESR) and Mossbauer spectroscopy. The enzyme "trypsin" was immobilized on the magnetic beads via adsorption. The maximum adsorption was obtained at pH 7.0. At 2.0 mg/mL initial trypsin concentration, the maximum immobilization capacity was 123.2 mg trypsin/g beads and retained about 84.2% of its initial activity. The immobilized trypsin could not be desorbed by enzyme reaction solution in the pH range of 5.0-9.0, and could be desorbed by 1.0 M formic acid solution containing 1 M NaCl. (C) 2007 Elsevier B.V. All rights reserved.
引用
收藏
页码:262 / 274
页数:13
相关论文
共 54 条
[11]   Covalent immobilization of chloroperoxidase onto magnetic beads: Catalytic properties and stability [J].
Bayramoglu, Guelay ;
Kiralp, Senem ;
Yilmaz, Meltem ;
Toppare, Levent ;
Arica, M. Yakup .
BIOCHEMICAL ENGINEERING JOURNAL, 2008, 38 (02) :180-188
[12]   Immobilization of β-galactosidase onto magnetic poly(GMA-MMA) beads for hydrolysis of lactose in bed reactor [J].
Bayramoglu, Gulay ;
Tunali, Yagmur ;
Arica, M. Yakup .
CATALYSIS COMMUNICATIONS, 2007, 8 (07) :1094-1101
[13]   Nanophase iron oxides by ball-mill grinding and their Mossbauer characterization [J].
Bid, S ;
Banerjee, A ;
Kumar, S ;
Pradhan, SK ;
De, UY ;
Banerjee, D .
JOURNAL OF ALLOYS AND COMPOUNDS, 2001, 326 (1-2) :292-297
[14]   ENZYME REACTION-ENGINEERING - DESIGN OF PEPTIDE-SYNTHESIS BY STABILIZED TRYPSIN [J].
BLANCO, RM ;
ALVARO, G ;
GUISAN, JM .
ENZYME AND MICROBIAL TECHNOLOGY, 1991, 13 (07) :573-583
[15]  
BOA H, 2006, MATER LETT, V60, P2167
[16]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[17]   Immobilisation of trypsin on acrylic copolymers [J].
Bryjak, J ;
Kolarz, BN .
PROCESS BIOCHEMISTRY, 1998, 33 (04) :409-417
[18]   Trypsin-based monolithic bioreactor coupled on-line with LC/MS/MS system for protein digestion and variant identification in standard solutions and serum samples [J].
Calleri, E ;
Temporini, C ;
Perani, E ;
De Palma, A ;
Lubda, D ;
Mellerio, G ;
Sala, A ;
Galliano, M ;
Caccialanza, G ;
Massolini, G .
JOURNAL OF PROTEOME RESEARCH, 2005, 4 (02) :481-490
[19]   Immobilization of catalase onto chitosan and cibacron blue F3GA attached chitosan beads [J].
Cetinus, Senay Akkus ;
Oztop, H. Nursevin ;
Saraydin, Dursun .
ENZYME AND MICROBIAL TECHNOLOGY, 2007, 41 (04) :447-454
[20]   Peptide bond formation by the industrial protease, neutrase, in organic media [J].
Clapes, P ;
Pera, E ;
Torres, JL .
BIOTECHNOLOGY LETTERS, 1997, 19 (10) :1023-1026