Chich J F, Gripon J C, Ribadeau-Dumas B
Unité Protéines, I.N.R.A., Station de Recherches Laitières, Jouy-en-Josas, France.
Anal Biochem. 1995 Jan 1;224(1):245-9. doi: 10.1006/abio.1995.1036.
To obtain large amounts of the X-prolyl dipeptidyl aminopeptidase from Lactococcus lactis subsp lactis (PepX, E.C. 3.4.14.5), PepX was purified from a commercial L. lactis cell extract. The enzyme was purified in only three steps and the last one was performed by HPLC on a C4 reverse-phase column using acetonitrile as an eluent. Despite its high molecular mass (175 kDa), the enzyme was recovered with a good activity yield (75%). Advantages and drawbacks of this technique compared to the classical ones are discussed. The stability of the enzyme in aqueous solutions and in the presence of 10 water-miscible solvents was also investigated. PepX was found to be stabilized by dimethyl sulfoxide, triglyme, and glycerol.
为了从乳酸乳球菌乳酸亚种（PepX，E.C. 3.4.14.5）中获得大量的X-脯氨酰二肽基氨基肽酶，从市售的乳酸乳球菌细胞提取物中纯化了PepX。该酶仅通过三步纯化，最后一步通过HPLC在C4反相柱上使用乙腈作为洗脱剂进行。尽管其分子量较高（175 kDa），但该酶的活性回收率良好（75%）。讨论了该技术与传统技术相比的优缺点。还研究了该酶在水溶液和10种与水混溶的溶剂存在下的稳定性。发现二甲基亚砜、三甘醇二甲醚和甘油可使PepX稳定。
