The magnetic chitosan nanoparticles were prepared by reversed-phase suspension method using Span-80 as an emulsifier, glutaraldehyde as cross-linking reagent. And the nanoparticles were characterized by TEM, FT-IR and hysteresis loop. The results show that the nanoparticles are spherical and almost superparamagnetic. The laccase was immobilized on nanoparticles by adsorption and subsequently by cross-linking with glutaraldehyde. The immobilization conditions and charac-terizations of the immobilized laccase were investigated. The optimal immobilization conditions were as follows: 10 mL of phosphate buffer (0.1 M, pH 7.0) containing 50 mg of magnetic chitosan nanoparticles, 1.0 mg·mL-1 of laccase and 1% (v/v) glutaraldehyde, immobilization temperature of 4 ℃ and immobilization time of 4 h. The immobilized laccase exhibited an appreciable catalytic capability (480 units·g-1 support) and had good storage stability and operation stability. The Km of immobilized and free laccase for ABTS were 140.6 and 31.1 μM in phosphate buffer (0.1 M, pH 3.0) at 37 ℃, respectively. The immobilized laccase is a good candidate for the research and development of biosensors based on laccase catalysis.
参考文献
| [1] | Setti Leonardo;Spinozzi Giovanni;Pifferi Pier Giogio;Giuliani Silvia .Laccase catalyzed-oxidative coupling of 3-methyl 2-benzothiazolinone hydrazone and methoxyphenols[J].Enzyme and Microbial Technology,1999(3/5):285-289. |
| [2] | Moeder M;Martin C;Koeller G .Degradation of hydroxylated compounds using laccase and horseradish peroxidase immobilized on microporous polypropylene hollow fiber membranes[J].Journal of Membrane Science,2004(1/2):183-190. |
| [3] | Osiadacz J.;Al-Adhami AJH.;Bajraszewska D.;Fischer P.;Peczynska-Czoch .On the use of Trametes versicolor laccase for the conversion of4-methyl-3-hydroxyanthranilic acid to actinocin chromophore[J].Journal of Biotechnology,1999(1/2):141-149. |
| [4] | D'Annibale A;Ricci M;Quaratino D;Federici F;Fenice M .Panus tigrinus efficiently removes phenols, color and organic load from olive-mill wastewater.[J].Research in Microbiology,2004(7):596-603. |
| [5] | Mousty C;Vieille L;Cosnier S .Laccase immobilization in redox active layered double hydroxides: A reagentless amperometric biosensor[J].Biosensors & Bioelectronics: The International Journal for the Professional Involved with Research, Technology and Applications of Biosensers and Related Devices,2007(8):1733-1738. |
| [6] | Patricio Peralta-Zamora;Claudia M.Pereira;Elaine R.L.Tiburtius;Sandra G.Moraes;Maria A.Rosa;Rosana C.Minussi;Nelson Duran .Decolorization of reactive dyes by immobilized laccase[J].Applied Catalysis, B. Environmental: An International Journal Devoted to Catalytic Science and Its Applications,2003(2):131-144. |
| [7] | Dominguez A;Gomez J;Lorenzo M;Sanroman A .Enhanced production of laccase activity by Trametes versicolor immobilized into alginate beads by the addition of different inducers.[J].World Journal of Microbiology & Biotechnology,2007(3):367-373. |
| [8] | Suna Timur;Nurdan Pazarlioglu;Roberto Pilloton;Azmi Telefoncu .Thick film sensors based on laccases from different sources immobilized in polyaniline matrix[J].Sensors and Actuators, B. Chemical,2004(1):132-136. |
| [9] | De-Sheng Jiang;Sheng-Ya Long;Jun Huang;Hai-Yan Xiao;Ju-Ying Zhou .Immobilization of Pycnoporus sanguineus laccase on magnetic chitosan microspheres[J].Biochemical Engineering Journal,2005(1):15-23. |
| [10] | K.Ravi Charan Reddy;Arvind M.Kayastha .Improved stability of urease upon coupling to alkylamine and arylamine glass and its analytical use[J].Journal of molecular catalysis, B. Enzymatic,2006(2):104-112. |
| [11] | Huang J;Xiao HY;Li B;Wang JT;Jiang DS .Immobilization of Pycnoporus sanguineus laccase on copper tetra-aminophthalocyanine-Fe3O4 nanoparticle composite[J].Biotechnology and Applied Biochemistry,2006(2):93-100. |
| [12] | Senay Akkus Cetinus;H.Nursevin Oztop .Immobilization of catalase into chemically crosslinked chitosan beads[J].Enzyme and Microbial Technology,2003(7):889-894. |
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