Abstract
Type II diabetes mellitus has been a global health problem with adverse side effects if left untreated. Recent studies have shown the inhibition of α-Amylase in degrading dietary starch is able to reduce postprandial hyperglycaemia in diabetic patients. Developments of specific inhibitors for α-Amylase thus provide an attractive alternative for the management of diabetes. Inhibiton of α-Amylase may be achieved by promoting inhibitor complexation by induced fit binding for enzyme inactivation. Peptides that are capable of mimicking or block the active sites would be idle inhibitors. Here, we employed peptide phage display to obtain a bioactive substitute capable of inhibiting α-Amylase activity. After three rounds of biopanning against α-Amylase using a 20-mer random peptide library, enrichment of target specific peptide pools were analyzed. Enriched pool of phage peptides were further used to screen for monoclones that are specific to α-Amylase. Two peptides, FAPMTSVMQSWWDVWRGGS (G5) and CLAEFTLWDRFWFGGRSGGR (G6) were identified from DNA sequencing. These two peptides at phage level exhibited approximately 18 % inhibition of α-Amylase activity. Purified G5 peptide was showed approximately 50 % inhibition for both Bacillus sp. and porcine pancreatic α-Amylase, but purified G6 peptide only presented approximately 40 % inhibition of Bacillus sp. α-Amylase. The use of phage display to identify peptides, which mimic the ‘natural’ motifs that contribute to the enzyme activity of α-Amylase may be a promising approach for management of diabetes.
This is a preview of subscription content, log in via an institution to check access.
References
Bulet P, Stöcklin R, Menin L (2004) Anti-microbial peptides: from invertebrates to vertebrates. Immunol Rev 198:169–184
Casey JL, Coley AM, Anders RF, Murphy VJ, Humberstone KS, Thomas AW, Foley M (2004) Antibodies to malaria peptide mimics inhibit Plasmodium falciparum invasion of erythrocytes. Infect Immun 72:1126–1134
Chen H-M, Muramoto K, Yamauchi F, Nokihara K (1996) Antioxidant activity of designed peptides based on the antioxidative peptide isolated from digests of a soybean protein. J Agric Food Chem 44:2619–2623
Davalos A, Miguel M, Bartolome B, Lopez-Fandino R (2004) Antioxidant activity of peptides derived from egg white proteins by enzymatic hydrolysis. J Food Prot 67:1939–1944
Deshayes K, Schaffer ML, Skelton NJ, Nakamura GR, Kadkhodayan S, Sidhu SS (2002) Rapid identification of small binding motifs with high-throughput phage display: discovery of peptidic antagonists of IGF-1 function. Chem Biol 9:495–505
Fowler MJ (2007) Diabetes treatment, Part 2: oral agents for glycemic management. Clin Diabetes 25:131–134
Hoogenboom HR, de Bruïne AP, Hufton SE, Hoet RM, Arends J-W, Roovers RC (1998) Antibody phage display technology and its applications. Immunotechnology 4:1–20
Hoskin DW, Ramamoorthy A (2008) Studies on anticancer activities of antimicrobial peptides. BBA-Biomembranes 1778:357–375
Kay BK (1995) Biologically displayed random peptides as reagents in mapping protein-protein interactions. Perspect Drug Discov Des 2:251–268
Lakshmanasenthil SVT, Geetharamani D, Maruthupandi T (2013) Screening of seaweeds collected from southeast coastal area of India for alpha amylase inhibitory activity, antioxidant activity and biocompatibility. Int J Pharm Pharm Sci 5:240–244
Lakshmanasenthil S, Vinothkumar T, Geetharamani D, Marudhupandi T, Suja G, Sindhu N (2014) Fucoidan—a novel α-amylase inhibitor from Turbinaria ornata with relevance to NIDDM therapy. Biocatal Agric Biotechnol 3:66–70
Mantzourani E, Laimou D, Matsoukas MT, Tselios T (2008) Peptides as therapeutic agents or drug leads for autoimmune, hormone dependent and cardiovascular diseases. Antiinflamm Antiallergy Agents Med Chem 7:294–306
Mendis E, Rajapakse N, Kim S-K (2005) Antioxidant properties of a radical-scavenging peptide purified from enzymatically prepared fish skin gelatin hydrolysate. J Agric Food Chem 53:581–587
Mitra A (2008) Some salient points in dietary and life-style survey of rural Bengal particularly tribal populace in relation to rural diabetes prevalence. Ethno Med 2:51–56
Pande J, Szewczyk MM, Grover AK (2010) Phage display: concept, innovations, applications and future. Biotechnol Adv 28:849–858
Peña-Ramos EA, Xiong YL, Arteaga GE (2004) Fractionation and characterisation for antioxidant activity of hydrolysed whey protein. J Sci Food Agric 84:1908–1918
Qian Z-J, Jung W-K, Kim S-K (2008) Free radical scavenging activity of a novel antioxidative peptide purified from hydrolysate of bullfrog skin, Rana catesbeiana Shaw. Bioresour Technol 99:1690–1698
Rajapakse N, Mendis E, Jung W-K, Je J-Y, Kim S-K (2005) Purification of a radical scavenging peptide from fermented mussel sauce and its antioxidant properties. Food Res Int 38:175–182
Russel M, Lowman HB, Clackson T (2004) Introduction to phage biology and phage display. In: Clackson T, Lowman HB (eds) Practical approach to phage display. Oxford University Press, New York, pp 1–26
Willats WG (2002) Phage display: practicalities and prospects. Plant Mol Biol 50:837–854
Worthington V (1993) Alpha amylase. In: Worthington V (ed) Worthington enzyme manual. Worthington Biochemical Corporation, Lakewood, pp 36–40
Acknowledgments
The authors would like to acknowledge support by the Research University Cluster Grant (Grant No. 1001/PSKBP/8630015).
Conflict of Interest
All authors declare no conflict of interest.
Statement of Informed Consent
Authors declare that there is no informed consent in the article.
Statement of Human and Animal Rights
This article does not contain any studies with human or animal subjects performed by any of the authors.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Chin, C.F., Tan, SJ., Gan, C.Y. et al. Identification of Peptide Based Inhibitors for α-Amylase by Phage Display. Int J Pept Res Ther 21, 237–242 (2015). https://doi.org/10.1007/s10989-015-9456-x
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10989-015-9456-x