MacLachlan J, Wotherspoon A T, Ansell R O, Brooks C J
Department of Physical Sciences, Glasgow Caledonian University, City Campus, 70 Cowcaddens Road, Glasgow, UK.
J Steroid Biochem Mol Biol. 2000 Apr;72(5):169-95. doi: 10.1016/s0960-0760(00)00044-3.
Since Flegg (H.M. Flegg, An investigation of the determination of serum cholesterol by an enzymatic method, Ann. Clin. Biochem. 10 (1973) 79-84) and Richmond (W. Richmond, The development of an enzymatic technique for the assay of cholesterol in biological fluids, Scand. J. clin. Lab. Invest. 29 (1972) 25; W. Richmond, Preparation and properties of a bacterial cholesterol oxidase from Nocardia sp. and its application to enzyme assay of total cholesterol in serum, Clinical Chemistry 19 (1973) 1350-1356) first illustrated the suitability of cholesterol oxidase (COD) for the analysis of serum cholesterol, COD has risen to become the most widely used enzyme in clinical laboratories with the exception of glucose oxidase (GOD). The use is widespread because assays incorporating the enzyme are extremely simple, specific, and highly sensitive and thus offer distinct advantages over the Liebermann-Burchard analytical methodologies which employ corrosive reagents and can be prone to unreliable results due to interfering substances such as bilirubin. Individuals can now readily determine their own serum cholesterol levels with a simple disposable test kit. This review discusses COD in some detail and includes the topics: (1) The variety of bacterial sources available; (2) The various extraction/purification protocols utilised in order to obtain protein of sufficient clarification (purity) for use in food/clinical analysis; (3) Significant differences in the properties of the individual enzymes; (4) Substrate specificities of the various enzymes; (5) Examples of biological assays which have employed cholesterol oxidase as an integral part of the analysis, and the various assay protocols; (6) New steroidal products of COD. This review is not a comprehensive description of published work, but is intended to provide an account of recent and current research, and should promote further interest in the application of enzymes to analytical selectivity.
自从弗莱格（H.M. 弗莱格，《酶法测定血清胆固醇的研究》，《临床生物化学年鉴》10（1973年）79 - 84页）和里士满（W. 里士满，《生物体液中胆固醇测定酶技术的发展》，《斯堪的纳维亚临床实验室研究杂志》29（1972年）25页；W. 里士满，《诺卡氏菌属细菌胆固醇氧化酶的制备及其性质及其在血清总胆固醇酶法测定中的应用》，《临床化学》19（1973年）1350 - 1356页）首次证明胆固醇氧化酶（COD）适用于血清胆固醇分析以来，除葡萄糖氧化酶（GOD）外，COD已成为临床实验室中使用最广泛的酶。其应用广泛是因为包含该酶的检测方法极其简单、特异且高度灵敏，因此相较于使用腐蚀性试剂且因胆红素等干扰物质容易产生不可靠结果的利伯曼 - 布查德分析方法具有明显优势。现在个人可以使用简单的一次性检测试剂盒轻松自行测定血清胆固醇水平。本综述详细讨论了COD，包括以下主题：（1）可用的多种细菌来源；（2）为获得用于食品/临床分析的足够澄清度（纯度）的蛋白质而采用的各种提取/纯化方案；（3）各个酶性质的显著差异；（4）各种酶的底物特异性；（5）将胆固醇氧化酶作为分析不可或缺部分的生物检测实例以及各种检测方案；（6）COD的新甾体产物。本综述并非已发表工作的全面描述，而是旨在介绍近期和当前的研究情况，并应激发对酶在分析选择性应用方面的进一步兴趣。
