Improved Method for Hydrolyzing Proteins and Peptides Without Inducing Racemization and for Determining Their True D-Amino Acid Content
✍ Scribed by A. Daniello; L. Petrucelli; C. Gardner; G. Fisher
- Publisher
- Elsevier Science
- Year
- 1993
- Tongue
- English
- Weight
- 503 KB
- Volume
- 213
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A new method of hydrolyzing proteins and peptides without racemizing the amino acids has been developed. This method consists of performing a brief partial chemical hydrolysis for (15 \mathrm{~min}) in (6 \mathrm{M} \mathrm{HCl}) at (80-90^{\circ} \mathrm{C}), followed by an enzymatic hydrolysis with pronase for (12-16 \mathrm{~h}) at (50^{\circ} \mathrm{C}), and finally an enzymatic hydrolysis with leucine aminopeptidase and peptidyl-D-amino acid hydrolase for (24 \mathrm{~h}). Using this new method the time required for complete hydrolysis of proteins is less than 3 days. The total hydrolysis averages (97-100 %), and the amount of racemization of the amino acids is less than (0.002 %). This method may then be used as a tool to easily determine the intrinsic (D)-amino acid content of peptides or proteins from animal or vegetable tissues. (c) 1993 Academic Press, Inc.