title: Development of a workflow for screening and identification of α-amylase inhibitory peptides from food source using an integrated Bioinformatics-phage display approach: Case study - Cumin seed.
abstract: The main objective of this study was to develop an efficient workflow to discover α-amylase inhibitory peptides from cumin seed. A total of 56 unknown peptides was initially found in the cumin seed protein hydrolysate. They were subjected to 2 different in silico screenings and 6 peptides were shortlisted. The peptides were then subjected to in vitro selection using phage display technique and 3 clones (CSP3, CSP4 and CSP6) showed high affinity in binding α-amylase. These clones were subjected to the inhibitory test and only CSP4 and CSP6 exhibited high inhibitory activity. Therefore, these peptides were chemically synthesized for validation purposes. CSP4 exhibited inhibition of bacterial and human salivary α-amylases with IC50 values of 0.11 and 0.04μmol, respectively, whereas CSP6 was about 0.10 and 0.15μmol, respectively. Results showed that the strength of each protocol has been successfully combined as deemed fit to enhance the α-amylase inhibitor peptide discovery.
authors: Analytical Biochemistry Research Centre, Universiti Sains Malaysia, 11800 USM, Penang, Malaysia.
authors: Hwee-Leng
authors: HL
authors: Siow
authors: Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 USM, Penang, Malaysia. Electronic address: theamsoon@usm.my.
authors: Theam Soon
authors: TS
authors: Lim
authors: Analytical Biochemistry Research Centre, Universiti Sains Malaysia, 11800 USM, Penang, Malaysia. Electronic address: cygan@usm.my.
authors: Chee-Yuen
authors: CY
authors: Gan
mesh_terms: D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000235:genetics / Q000378:metabolism; D019295:Computational Biology / Q000379:methods*; D028527:Cuminum / Q000235:genetics / Q000378:metabolism; D006801:Humans; D010446:Peptide Fragments / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D012639:Seeds / Q000235:genetics / Q000378:metabolism; D057188:Workflow; D000516:alpha-Amylases / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D010446:Peptide Fragments; D000516:alpha-Amylases
keywords: Bioactive peptide discovery; Bioinformatics; Phage display; α-Amylase inhibition