Papaya latex enzymes capable of detoxification of gliadin.
Assay of fractions obtained from ion exchange chromatography of papaya latex on CM Sephadex-C50, size exclusion chromatography on Sephacryl S-300 and size exclusion HPLC have provided an insight into the relative contributions of the gluten-detoxifying enzymes present. This outcome has been achieved by the use of the above chromatographic techniques, coupled with assays of lysosomal activity, protease activity using benzylarginine ethyl ester (BAEE) as substrate, prolyl endopeptidase (PEP) using glycylprolylnitroanilide and a prolidase assay using acetylprolylglycine. These procedures have shown that the activity in papaya latex is due largely to caricain and to a lesser extent, chymopapain and glutamine cyclotransferase. The presence of caricain and these other enzymes was confirmed by mass spectrometry of trypsin digests of the most active fraction obtained by CM Sephadex-C50 chromatography and size exclusion HPLC. Fractions rich in caricain would be suitable for enzyme therapy in gluten intolerance and appear to have synergistic action with porcine intestinal extracts.
Cornell
H J
HJ
School of Applied Sciences, RMIT University, Melbourne, VIC, 3001, Australia. hugh.cornell@rmit.edu.au
Doherty
W
W
Stelmasiak
T
T
D019881:Aminoacyltransferases / Q000737:chemistry* / Q000302:isolation & purification; D029441:Carica / Q000737:chemistry / Q000201:enzymology*; D002384:Catalysis; D002446:Celiac Disease / Q000378:metabolism; D002917:Chymopapain / Q000737:chemistry* / Q000302:isolation & purification; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification; D005903:Gliadin / Q000737:chemistry* / Q000633:toxicity; D006801:Humans; D007700:Kinetics; D007840:Latex / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification
D007840:Latex; D010940:Plant Proteins; D005903:Gliadin; D019881:Aminoacyltransferases; C059280:glutaminyl-peptide cyclotransferase; D003546:Cysteine Endopeptidases; C039974:caricain; D002917:Chymopapain