A Comparison of Assays on Hydrolytic Activity of Lipase with and without Surfactant
A Comparison of Assays on Hydrolytic Activity of Lipase with and without Surfactant
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有表面活性剂和无表面活性剂的脂肪酶水解活性测定的比较
DOI：
10.5650/jos1956.36.402
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
T. Yamane
中科院分区：
文献类型：
--
作者：
T. Yamane
脂肪酶（三酰甘油酰基水解酶，EC 3.1。1.3)是一种普遍存在的酶，其催化脂肪酸从其甘油酯水解。虽然脂肪酶是油脂消化的重要酶之一，但相对于淀粉酶、蛋白酶等水解酶，对脂肪酶的研究较少。然而，鉴于对油脂工业的生物技术的兴趣日益增加，脂肪酶催化的水解（脂肪分解）或脂质的合成已经变得比学术兴趣更重要。脂肪酶催化的其它反应如酸解、醇解、酯交换和外消旋化合物的旋光拆分等也引起了学术界和工业界的广泛关注，因为这些生物转化有望从脂肪、油脂、脂肪酸及其相关化合物中生产出各种高附加值的产品。几种微生物脂肪酶现在以合理的价格可商购获得，并且为了上述目的正在开发具有独特性质的新脂肪酶。脂肪酶催化的任何生物过程，都需要一种可靠的测定其活性的方法来评价生物过程的可行性。精确定量测定其活性是研究和开发涉及酶的生物过程的基础。合适的活性分析方法是监测纯化和鉴定特异性的先决条件。早在20世纪40年代，许多研究者就发展了各种测定脂肪酶活力的方法。1983年报道的大多数分析以及检测脂肪酶的方法由Jensen综述。一年后，又出现了几篇关于脂肪酶活性测定的论文。脂肪酶水解活性的测定方法按底物的种类大致可分为水溶性底物法和水不溶性底物法。作为水溶性底物，使用三醋精（三乙酰甘油）、三丁酸甘油酯（三丁酸甘油）或S-酰基化合物（三丁酸-1，2-二硫代甘油，BALB）。最后一种化合物用于测定血清中脂肪酶活性，使用BALB的试剂盒正在商业化。另一方面，橄榄油、三油酸甘油酯（三油酰甘油）或显色底物如4-甲基香豆素的脂肪酸酯和三[12-（2，4，6 *）生物反应器用于脂肪和油，第1X部分，对于第1部分-VU，参见J. Jpn Oil Chem. Sac.
Lipase (triacylglycerol acylhydrolase, EC 3.1. 1.3) is an ubiquitous enzyme which catalyzes the hydrolysis of fatty acids from their glycerol esters. Although the enzyme is one of the important enzymes for lipid digestion, studies on lipase had been less than those on the other hydrolytic enzymes such as amylases and prow teases. However, in view of the increasing interest in biotechnology for the fats and oils industry, lipase-catalyzed hydrolysis (fat splitty ing) or synthesis of lipids has become of more than academic interest. Also, some other reacv tions catalyzed by lipase such as acidolysis, alcoholysis, interesterification and optical rev solution of racemic compounds have recently attracted much attention of both academic and industrial researchers because these bioconverv sion are expected to produce various valueadded products from fats, oils, fatty acids and their related compounds. Several microbial lipases are now available commercially with reasonable prices, and new lipases having unique properties are being developed for the purposes mentioned above. Whatever bioprocv ess catalyzed by lipase is concerned, a reliable method of measurement of its activity is needed to evaluate the feasibility of the bioprocess. Precise quantitative determination of its activity is essential as the basis of research and develop ment of the bioprocess involving the enzyme. Suitable methods for analysis of activity are prerequisites for monitoring purification and identification of specificity. Since as early as 1940 s, many investigators have developed various methods for the deter= mination of lipase activity. Most of these anv alyses as well as procedures for detection of lipases reported by the year 1983 was reviewed by Jensenl'. Still after the year, several papers appeared on the determination of lipase acv tivity2'"°. The assay methods of hydrolytic activity of lipases are broadly classified into two with re= spect to the kind of substrate used, the ones using water-soluble substrates and the ones using water-insoluble substrates. As watersoluble substrates, triacetin (triacetylglycerol), tributyrin (tributyrylglycerol) or S-acyl com= pound (tributyryl-1, 2-dithioglycerol, BALB) is used. The last compound is used for the deterv urination of lipase activity in serum and a kit using BALB is being comercialized. On the other hand, olive oil, triolein (trioleoylglycerol) or chromogenic substrates such as fatty acid esters of 4-methylcoumarin and tris [12-(2, 4, 6* Bioreactors for Fats and Oils , Part 1X, For Parts 1-VU, see J. Jpn Oil Chem. Sac.