Kurooka S, Kitamura T
J Biochem. 1978 Dec;84(6):1459-66. doi: 10.1093/oxfordjournals.jbchem.a132269.
Very low levels of lipase can easily be measured by a new serum lipase assay method (the BALB-DTNB method), using BAL-tributyrate (BALB) as a substrate, 5,5'-dithiobis(2-nitrobenzoic acid) as a chromogenic SH reagent, phenylmethylsulfonylfluoride as an inhibitor of esterases and sodium dodecyl sulfate as a surfactant. The BALB-DTNB method has a higher sensitivity than the conventional serum lipase assay methods, and proved useful for analyzing the properties of serum lipases in combination with gel-filtration on a Sephacryl S 200 column and isoelectrofocusing in an Ampholine column. Serum samples containing high levels of lipases from patients with pancreatic diseases or patients in whom the pancreatic exocrine gland had been stimulated by injecting caerulein and secretin were analyzed by these methods. The lipolytic profiles obtained indicated the presence of a lipase with an estimated molecular weight of 46,000 and isoelectric points of 7.4, 6.8, or/and 6.4. A lipase with properties similar to those of the serum lipase was found to be present in human pancreatic juice.
采用一种新的血清脂肪酶检测方法(BAL - DTNB法),以丁酰硫代胆碱(BALB)为底物、5,5'-二硫代双(2 - 硝基苯甲酸)为显色巯基试剂、苯甲基磺酰氟为酯酶抑制剂、十二烷基硫酸钠为表面活性剂,能够轻松检测出极低水平的脂肪酶。BALB - DTNB法比传统的血清脂肪酶检测方法具有更高的灵敏度,并且已证明该方法结合在Sephacryl S 200柱上的凝胶过滤和在两性电解质柱上的等电聚焦,对于分析血清脂肪酶的性质很有用。通过这些方法对患有胰腺疾病患者或通过注射蛙皮素和促胰液素刺激胰腺外分泌腺的患者的含高水平脂肪酶的血清样本进行了分析。获得的脂解图谱表明存在一种估计分子量为46,000且等电点为7.4、6.8或/和6.4的脂肪酶。发现人胰液中存在一种性质与血清脂肪酶相似的脂肪酶。