Recovery of tryptophan from 25-minute acid hydrolysates of protein.
It was found that thioglycolic acid prevents destruction of tryptophan during rapid hydrolysis of protein with a trifluoroacetic acid/HCl mixture (1:2, v/v) at 166 degrees C for 25 or 50 min. The addition of 5% (v/v) thioglycolic acid gave the maximum tryptophan recovery (88.3%) for a 25-min hydrolysate of lysozyme. Tryptophan recoveries varied slightly among three different proteins; 88% for lysozyme, 73% for alpha-chymotrypsinogen A, and 85% for apomyoglobin. However, when extrapolated to zero time, the values were close to one another: 94, 87, and 88%, respectively. The addition of thioglycolic acid was also advantageous for recovering amino acids other than tryptophan. Particularly, yields of carboxymethylcysteine and methionine were greatly improved. This modified rapid hydrolysis method gave satisfactory results without the need for separate analyses of tryptophan and cysteine, provided proteins were reduced and carboxymethylated prior to hydrolysis.
D000143:Acids; D000596:Amino Acids / Q000032:analysis; D001059:Apoproteins; D002919:Chymotrypsinogen / Q000302:isolation & purification; D006868:Hydrolysis; D009113:Muramidase / Q000302:isolation & purification; D009211:Myoglobin / Q000302:isolation & purification; D010446:Peptide Fragments / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification*; D013864:Thioglycolates; D013997:Time Factors; D014364:Tryptophan / Q000302:isolation & purification*
D000143:Acids; D000596:Amino Acids; D001059:Apoproteins; D009211:Myoglobin; D010446:Peptide Fragments; D011506:Proteins; D013864:Thioglycolates; C010857:apomyoglobin; D014364:Tryptophan; D002919:Chymotrypsinogen; D009113:Muramidase