Recovery of tryptophan in peptides and proteins by high-temperature and short-term acid hydrolysis in the presence of phenol.
The addition of 3% (w/v) phenol to 6 M HCl largely prevented the destruction of tryptophan during rapid hydrolysis of peptides and proteins at 166 degrees C for 25 min or at 145 degrees C for 4 h. This hydrolysis procedure was advantageous for amino acid microanalysis using conventional high-performance liquid chromatography with a precolumn derivatization technique. The recovery of tryptophan from proteins was at least 80%. The addition of phenol also improved the recovery of methionine and carboxymethylcysteine. The amount of tryptophan in proteins electroblotted onto a polyvinylidene difluoride membrane was determined by this method.
Muramoto
K
K
School of Fisheries Sciences, Kitasato University, Iwate, Japan.
Kamiya
H
H
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003064:Cnidarian Venoms / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D037102:Lectins / Q000032:analysis; D008567:Membranes, Artificial; D010455:Peptides / Q000378:metabolism*; D019800:Phenol; D010636:Phenols / Q000494:pharmacology; D011145:Polyvinyls; D011506:Proteins / Q000378:metabolism*; D012394:Rosaniline Dyes; D012694:Serine / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate; D013912:Threonine / Q000378:metabolism; D013997:Time Factors; D014364:Tryptophan / Q000378:metabolism*; D004124:p-Dimethylaminoazobenzene / Q000378:metabolism / Q000031:analogs & derivatives; D005465:Fluorocarbon Polymers
D003064:Cnidarian Venoms; D037102:Lectins; D008567:Membranes, Artificial; D010455:Peptides; D019800:Phenol; D010636:Phenols; D011145:Polyvinyls; D011506:Proteins; D012394:Rosaniline Dyes; D012694:Serine; D012967:Sodium Dodecyl Sulfate; D013912:Threonine; D014364:Tryptophan; D004124:p-Dimethylaminoazobenzene; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers; C010572:4-N,N-dimethylaminoazobenzene-4'-sulfonyl chloride; C036310:toxin I (Anemonia sulcata); C024757:Coomassie brilliant blue R