Preparation of Bacterial X-Prolyl Dipeptidyl Aminopeptidase and Its Stabilization by Organic Cosolvents
✍ Scribed by J.F. Chich; J.C. Gripon; B. Ribadeaudumas
- Publisher
- Elsevier Science
- Year
- 1995
- Tongue
- English
- Weight
- 418 KB
- Volume
- 224
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
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 (C_{4}) reverse-phase column using acetonitrile as an eluent. Despite its high molecular mass ( (175 \mathrm{kDa}) ), the enzyme was recovered with a good activity yield ((\mathbf{7 5 %})). 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. 1995 Academic Press, Inc.