新型方法制备固定化脲酶基质材料及固定化脲酶的性能研究Hydrothermal Synthesis of Immobilized Urease Matrix Materials and Exploration the Performance of Immobilized Urease
郭明,燕冰宇,杨萍
摘要(Abstract)：
以微晶纤维素(Microcrystalline cellulose,MCC)为原料,采用新型水热氧化方法,制备纤维素基质固定化脲酶载体材料-双醛纤维素(Dialdehyde cellulose,DAC)。采用红外光谱(IR)、固体核磁共振(CP/MAS 13C NMR)、X-射线衍射(XRD)和扫描电镜(SEM)表征产物结构。测试制备的纤维素基质固载脲酶的酶学性能,并与交联壳聚糖、双醛淀粉固载脲酶的酶学性能进行比较分析,构建酶学性能理论方程。结果表明:采用新型水热氧化反应可成功制备高醛基含量的氧化纤维素,与交联壳聚糖和双醛淀粉固定化脲酶相比,所得氧化纤维素载体材料固定化脲酶的米氏常数小(0.0108mol·L-1),对底物亲和能力强,重复使用率高,在pH=5.0~9.0能保持酶活性,建立的理论方程较好地表征了固载酶酶学性能。
关键词(KeyWords)： 双醛纤维素;水热合成;固定化脲酶;结构表征;酶学性能
基金项目(Foundation): 浙江省自然科学基金项目(Y5090179);; 浙江省科技厅重大科技专项项目(2010C33070
作者(Author): 郭明,燕冰宇,杨萍
参考文献(References)：
- [1]Yuan L I,Emiko R,Selvi T,et al.Detection of clarithromycin resistance in Helicobacter pylori following noncryogenic storage of rapid urease tests for 30 days[J].Digestive Diseases,2012,13(1):54-59.
- [2]Yang Z P,Si S H,Zhang C J.Study on the activity and stability of urease immobilized onto nanoporous alumina membranes[J].Micropor Mesopor Mat,2008,111(1-3):359-366.
- [3]Maja H,Mateja P,Zeljko K.Enzymatic reactions in high-pressure membrane reactors[J].Ind Eng Chem Res,2005,44(25):9619-9625.
- [4]Vassar R.BACE1,the alzheimer's beta-secretase enzyme,in health and disease[J].Molecular Neurodegeneration,2004,23(1-2):105-114.
- [5]Krajewska B.Ureases II properties and their customizing by enzyme immobilizations:A review[J].Mol Catal B-Enzym,2009,59(1-3):22-40.
- [6]Cao L Q.Immobilised enzymes:science or art[J].Curr Opin Chem Biol,2005,9(2):217-226.
- [7]Noriyuki I,Lee D S,Kwon Y J,et al.Immobilization of protein on cellulose hydrogel[J].Cellulose,2011,18(5):1251-1256.
- [8]Opwis K,Knittel D,Schollmeyer E.Immobilizationof catalase on textile carrier materials[J].AATCC Review,2004,22(12):25-28.
- [9]Dey G,Nagpal V,Banerjee R.Immobilization ofα-amylase from bacillus circulans GRS 313 on cocount fiber[J].Applied Biochemistry and Biotechnology,2002,24(12):303-314.
- [10]Sirvio J,Liimatainen H,Niinimaki J,et al.Dialdehyde cellulose microfibers generated from wood pulp by milling-induced periodate oxidation[J].Carbohydrate Polymers,2011,86(1):260-265.
- [11]Wang Y H,Hsieh Y L.Aldehyde functionalized cellulose support for hydrogels[J].Journal of Applied Polymer Science,2010,118(5):2489-2495.
- [12]Varma A J,Kulkarni M P.Oxidati on of cellulose under controlled conditions[J].Polymer Degradation and Stability,2002,77(1):25-27.
- [13]Rahn K,Heinze L.Cellulosic polymers by subsequent modification of 2,3-dialdehyde cellulose[J].Cellulose Chemistry and Technology,1998,32(3):173-183.