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Identification of Prolyl Endopeptidase (PEP) Inhibitory Peptides from Lactobacillus helveticus Digestion of Four Recombinant Bovine Caseins
Author(s) -
Vishram Avleen,
Clack Beatrice
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.894.12
Subject(s) - casein , recombinant dna , lactobacillus helveticus , biochemistry , endopeptidase , cloning (programming) , biology , peptide , gluten , chemistry , enzyme , lactobacillus , microbiology and biotechnology , gene , programming language , fermentation , computer science
Eurygaster integriceps Puton (Sunn pest) is the most damaging insect pest of wheat in areas of North Africa, Eastern Europe and Western and Central Asia. Sunn pest injects an enzyme characterized as a prolyl endylprotease (spPEP) into wheat grains that digests the gluten protein (Darkoh et al., 2010). Previous studies have shown that casein peptides generated by Lactobacillus species have inhibitory effects on mammalian and bacterial prolyl endoproteases (PEP) which makes casein a good natural candidate against spPEP. This study used four purified recombinant bovine casein genes, α1, α2, β and κ to identify which Lactobacillus generated peptides will inhibit the spPEP. Firstly, these four recombinant casein genes were expressed and purified. Then the peptides were generated by incubating each purified casein with Lactobacillus helveticus . These peptides were separated using High Performance Liquid Chromatography (HPLC). The inhibitory peptides were identified by incubating them with recombinant PEP enzyme in the presence the GPpNA substrate. Mass and sequence of the inhibitory peptides were determined using MALDI Mass Spectrophotometry.. Future studies will include the cloning and expression of the identified peptides of different lengths to determine specificity unique to spPEP. Funding: Office of Research and Sponsored Projects, SFASU and Ed and Gwen Cole, Nacogdoches, TX.