Laboratorio de Investigacion de Proteinas Vegetales (LIPROVE), Depto. Cs. Biologicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina.
Prep Biochem Biotechnol. 2011;41(2):107-21. doi: 10.1080/10826068.2011.544230.
Papain from latex of Carica papaya was purified up to matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry homogeneity by salt precipitation from two different crude extract sources: a refined preparation obtained in our laboratory and a commercial one. Sodium tetrathionate was tested in the purification process to preserve the enzymatic activity of the peptidase. Purification was checked by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and cation exchange chromatography, using commercial pure papain as standard for a rapid comparison. The best purification yields (3.4%) were obtained in presence of 30 mM sodium tetrathionate for the crude extract prepared in our laboratory. The described purification method proved to be robust and reliable to obtain pure papain on a preparative scale.
木瓜蛋白酶从卡拉胶乳中被纯化到基质辅助激光解吸/电离(MALDI)飞行时间(TOF)质谱均一性,通过从两种不同的粗提物来源进行盐沉淀来实现:一种是在我们实验室中获得的精制制剂,另一种是商业制剂。四硫代硫酸钠在纯化过程中被测试以保持肽酶的酶活性。通过十二烷基硫酸钠(SDS)聚丙烯酰胺凝胶电泳(PAGE)和阳离子交换层析来检查纯化情况,使用商业纯木瓜蛋白酶作为标准进行快速比较。在我们实验室制备的粗提物中,当存在 30mM 四硫代硫酸钠时,获得了最佳的纯化产率(3.4%)。所描述的纯化方法被证明是稳健和可靠的,可在制备规模上获得纯木瓜蛋白酶。