共价色谱法。从干燥的木瓜乳胶中制备全活性木瓜蛋白酶。
Covalent chromatography. Preparation of fully active papain from dried papaya latex.
作者信息
Brocklehurst K, Carlsson J, Kierstan M P, Crook E M
出版信息
Biochem J. 1973 Jul;133(3):573-84. doi: 10.1042/bj1330573.
Abstract
- A Sepharose-(glutathione-2-pyridyl disulphide) conjugate has been prepared. 2. Its use in a new type of chromatography, covalent chromatography by thiol-disulphide interchange, is described. 3. With this technique, papain containing 1 intact catalytic site [thiol with high reactivity towards 2,2'-dipyridyl disulphide (2-Py-S-S-2-Py) at pH4] per mol of protein is readily prepared both from dried papaya latex and from commercial 2xcrystallized partially active papain. 4. The catalysis of the hydrolysis of alpha-N-benzoyl-l-arginine ethyl ester at pH6.0, 25.0 degrees C, I=0.3 by fully active papain thus prepared is characterized by K(m)=18.2+/-<0.1mm and k(cat.)=16.4+/-0.5s(-1).
摘要
- 已制备出一种琼脂糖-（谷胱甘肽-2-吡啶二硫化物）共轭物。2. 描述了其在一种新型色谱法——通过硫醇-二硫化物交换进行的共价色谱法中的应用。3. 利用该技术，每摩尔蛋白质含有1个完整催化位点[在pH4时对2,2'-二吡啶二硫化物（2-Py-S-S-2-Py）具有高反应活性的硫醇]的木瓜蛋白酶可轻松地从干燥的木瓜乳胶和市售的2x结晶部分活性木瓜蛋白酶中制备出来。4. 如此制备的完全活性木瓜蛋白酶在pH6.0、25.0℃、离子强度I = 0.3时对α-N-苯甲酰-L-精氨酸乙酯水解的催化作用的特征为米氏常数K(m)=18.2±0.1mM，催化常数k(cat.)=16.4±0.5s(-1)。
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Covalent chromatography. Preparation of fully active papain from dried papaya latex.共价色谱法。从干燥的木瓜乳胶中制备全活性木瓜蛋白酶。
Biochem J. 1973 Jul;133(3):573-84. doi: 10.1042/bj1330573.
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A necessary modification to the preparation of papain from any high-quality latex of Carica papaya and evidence for the structural integrity of the enzyme produced by traditional methods.对从任何优质番木瓜乳胶制备木瓜蛋白酶的必要改进以及传统方法生产的酶结构完整性的证据。
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KINETICS OF PAPAIN-CATALYZED HYDROLYSIS OF ALPHA-N-BENZOYL-L-ARGININE ETHYL ESTER AND ALPHA-N-BENZOYL-L-ARGININAMIDE.木瓜蛋白酶催化α-N-苯甲酰-L-精氨酸乙酯和α-N-苯甲酰-L-精氨酰胺水解的动力学
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Crystalline papain. I. Preparation, specificity, and activation.结晶木瓜蛋白酶。I. 制备、特异性及激活
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A novel reactivity of papain and a convenient active site titration in the presence of other thiols.木瓜蛋白酶的一种新反应活性以及在其他硫醇存在下简便的活性位点滴定法。
FEBS Lett. 1970 Jul 29;9(2):113-116. doi: 10.1016/0014-5793(70)80327-1.
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Chemical coupling of proteins to agarose.蛋白质与琼脂糖的化学偶联。
Nature. 1967 Sep 30;215(5109):1491-2. doi: 10.1038/2151491a0.
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Chemical coupling of peptides and proteins to polysaccharides by means of cyanogen halides.通过卤化氰将肽和蛋白质与多糖进行化学偶联。
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Determination of sulfhydryl groups with 2,2'- or 4,4'-dithiodipyridine.用2,2'-或4,4'-二硫代二吡啶测定巯基
Arch Biochem Biophys. 1967 Mar;119(1):41-9. doi: 10.1016/0003-9861(67)90426-2.
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The determination of the concentration of hydrolytic enzyme solutions: alpha-chymotrypsin, trypsin, papain, elastase, subtilisin, and acetylcholinesterase.水解酶溶液浓度的测定：α-胰凝乳蛋白酶、胰蛋白酶、木瓜蛋白酶、弹性蛋白酶、枯草杆菌蛋白酶和乙酰胆碱酯酶。
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