Improved method for hydrolyzing proteins and peptides without inducing racemization and for determining their true D-amino acid content

Anal Biochem. 1993 Sep;213(2):290-5. doi: 10.1006/abio.1993.1423.

Abstract

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 min in 6 M HCl at 80-90 degrees C, followed by an enzymatic hydrolysis with pronase for 12-16 h at 50 degrees C, and finally an enzymatic hydrolysis with leucine aminopeptidase and peptidyl-D-amino acid hydrolase for 24 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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / analysis*
  • Animals
  • Fabaceae
  • Hydrolysis
  • Liver / chemistry
  • Molecular Sequence Data
  • Octopodiformes
  • Peptides / analysis*
  • Plant Proteins / analysis
  • Plants, Medicinal
  • Proteins / analysis*
  • Stereoisomerism

Substances

  • Amino Acids
  • Peptides
  • Plant Proteins
  • Proteins