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Improvement of the extraction of penicillin acylase from Escherichia coli cells by a combined use of chemical methods

✍ Scribed by Isabel S. Novella; Claire Fargues; Georges GréVillot

Publisher
John Wiley and Sons
Year
1994
Tongue
English
Weight
396 KB
Volume
44
Category
Article
ISSN
0006-3592

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✦ Synopsis

Several techniques for protein extraction were tested for recovering penicillin acylase from a recombinant strain of Escherichia coli. These techniques include chemical [guanidine hydrochloride, Triton X-100, ethylenediaminetetraacetic acid (EDTA), ethanol/toluene], physical (sonication, freeze-and-thawing), and enzymatic (lysozyme) treatments. Best results were obtained with the combined use of guanidine and EDTA. This extraction procedure was optimized, and it was found that 95% of the enzyme was extracted after a 10 mM EDTA plus 10 mM guanidine treatment at room temperature for 10 h. The purification factor was 25 when compared to disruption by sonication. This extraction method could avoid purification Steps for particular applications.

📜 SIMILAR VOLUMES

Chemical treatment of Escherichia coli:
Chemical treatment of Escherichia coli: 3. Selective extraction of a recombinant protein from cytoplasmic inclusion bodies in intact cells
✍ Robert J. Falconer; Brian K. O'Neill; Anton P. J. Middelberg 📂 Article 📅 1999 🏛 John Wiley and Sons 🌐 English ⚖ 308 KB 👁 2 views

In previous parts of this study we developed procedures for the high-efficiency chemical extraction of soluble and insoluble protein from intact Escherichia coli cells. Although high yields were obtained, extraction of recombinant protein directly from cytoplasmic inclusion bodies led to low product