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中文摘要：
用简并引物PCR和5'-RACE从苛求芽孢杆菌ATCC26904中克隆出其胞内尿酸酶编码序列(FJ393559)并通过原核重组表达确认。此重组表达尿酸酶的米氏常数、对黄嘌呤抑制作用的抗性、最适pH和最适反应温度等同天然酶一致，用于动力学方法测定血清尿酸对血清黄嘌呤有很好抗性且线性范围宽。此酶等电点等电点接近5.0，有322个氨基酸，有活性的重组表达尿酸酶为同四聚体；此酶N端序列对其功能有显著影响，N端缺失可导致不能形成活性四聚体；增强N端和羧基端的离子键相互作用可提高其在pH 7.4时的热稳定性；同源建模发现此酶羧基端有伸展的肽段。此重组表达尿酸酶经20倍过量聚乙二醇5000-N-羟基琥珀酰亚胺酯修饰后，在37度pH 7.4时热失活半衰期延长约1倍；此酶在pH 9.0时热失活半衰期可长达5天。PEG修饰前后都可在离体的血浆中有效清除尿酸，PEG修饰后在健康大鼠体内血浆循环半衰期可达36小时，有药用潜力。
结论摘要：
英文主题词Uricase; cloning and recombinant expression; thermo-stability; PEGylation
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期刊论文
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Characterization of an intracellular uricase from Bacillus fastidious ATCC 26904 and its application
A new linearly-combined bi-exponential model for kinetic analysis of the isometric relaxation proces
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Soluble expression in E. coli of active human cyclic nucelotide phosphodiesterase isoform 4B 2 in fu
The measurement of cyclic nucleotide phosphodiesterase 4 activities via the quantification of inorga
An integration strategy to estimate the initial rates of enzyme reactions with expanded linear range
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