Hide
Show all
Email this article 
Bioactivities and analysis of peptides from Sumbawa horse milk generated by Bacillus thuringiensis protease
Abstract
Sumbawa horse milk is claimed to cure some diseases such as asthma, hypertension, diabetes and gastrointestinal disorder but its potential bioactive peptide has not been explored. The aims of this study are to evaluate bioactivities peptides from Sumbawa horse milk protein hydrolysate and to analyzethe physio-chemical properties of selected peptides. The milk protein was hydrolyzed by Bacillus thuringiensis protease, the peptide produced were sequential fractionated and then analyzed for antibacterial and antioxidant activities. The peptide fraction <3 kDa was then sequenced using LCMS-MS and the physio-chemical properties of the peptides were analyzed. The result showed that peptides fraction <3 kDa from the 30 min hydrolysis was the most active as antibacterial and more active to Gram negative bacteria. For antioxidant, scavenging activity of the fraction per µg protein/mL were 83% to ABTS and 31% to DPPH radicals. The values were similar with vitamin C 12.5 µg/mL for ABTS and 14.5 µg/mL for DPPH. Peptide HPYFYAPELLYYANK with molecular weight prediction 1887.92 Da and isoelectric point 7.47 has high therapeutic index prediction (64.75). The result showed that peptides from Sumbawa horse milk hydrolyzed by Bacillus thuringiensis protease was active as antibacterial and antioxidant. Peptide HPYFYAPELLYYANK from fraction <3 kDa was potential as antibacterial.
Â
Keywords
Full Text:
PDFReferences
Argolo-Filho RC, Loguercio LL. 2014. Bacillus thuringiensis is an environmental pathogen and host-specificity has developed as an adaptation to human-generated ecological niches. Insect. 5:62-91.
Abraham T, Prenner EJ, Lewis RNAH, Mant CT, Keller S, Hodges RS, Melhaney RN. 2014. Structure-activity relationships of the antimicrobial peptide gramicidin S and its analogs: Aqueous solubility, self-association, conformation, antimicrobial activity and interaction with model lipid membranes. Biochimica et Biophysica Acta. 1838:1420-1429.
Agastya AS, Sharma N, Mohan A, Mahal P. 2013. Isolation and molecular characterization of alkaline protease producing Bacillus thuringiensis. Cell Biochem Biophys. 66:45-51.
Ammons DR. 2009. An investigation of Bacillus thuringiensis in rectal-collected fecal samples of cows. Curr Microbiol. 59:532-536.
Amza T, Balla A, Tounkara F, Man L, Zhou HM. 2013. Effect of hydrolysis time on nutritional, functional and antioxidant properties of protein hydrolysates prepared from gingerbread plum (Neocarya macrophylla) seeds. Int Food Res J. 20:2081-2090.
Bahnsen JS, Franzyk H, Sandberg-Schaal A, Nielsen HM. 2013. Antimicrobial and cell-penetrating properties of penetratin analogs: Effect of sequence and secondary structure. Biochemica et Biophysica Acta. 1928:223-232.
Bai Y, Liu S, Li J, Laksminarayan R, Sarawathi P, Tang C, Ho D, Verma C, Beuerman RW, Pervushin K. 2012. Progressive structuring of a branced antimicrobial peptide on the path to inner membrane target. J Biol Chem. 287(32): 26606-26617.
Banerjee D, Gohil T, Trivedi S. 2015. Antibacterial activity of oliomeric β-defensin isolated from heart tissue of Capra hircus. Int J Pharm Sci. 7:131-135.
Barboza-Corona, JE, Vazquez-Acosta H, Bideshi DK, Salcedo-Hernandez R. 2007. Bacteriocin-like inihibitor substances produced by Mexican strains of Bacillus thuringiensis. Arch Microbiol. 187:117-126.
Bergmeyer HU, Bergmeyer J, Grassl M. 1983. Methods of enzymatic analysis. Vol 2. Weinheim: Verlag Chemie, Weinheim, Germany.
Chen Y, Guarnieri MT, Vasil AI, Vasil ML, Mant CT, Hodges RS. 2007. Role of peptide hydrophobicity in the mechanism of action of α-helical antimicrobial peptide, Antimicrob Agent Chemother. 51:1398-1406..
Correa AP, Daroit DJ, Coelho J, Meira SM, Lopes FC, Segalin J, Risso PH, Brandelli A. 2011. Antioxidant, antihypertensive and antimicrobial properties of ovine caseinate hydrolyzed with microbial protease. J Sci Food Agric. 91:2247-2254.
Damgaard TD, Otte JAH, Meinert L, Jensen K, Lametsch R. 2014. Antioxidant capacity of hydrolyzed porcine tissues. Food Sci Nutr. 2:282-288.
Detha A, Sudarwanto M, Latif H, Datta FU, Rahayu P. 2013. Fractionation and identification antimicrobial activity of Sumbawa mare milk protein against causative agent of subclinical mastitis. Global Vet. 11:674-668.
Domnez N, Kisadere I, Balaban C, Kadiralieva N. 2014. Effect of traditional homemade koumiss on some hematological and biochemical characteristics in sedentary men exposed to exercise. Biotech Histochem. 89:1-6.
Fernandez-Vidal M, Jayasinghe S, Ladokhin AS, White SH. 2007. Folding amphipathic helices into membranes: Amphiphilicity trumphs hydrophobicity. J Mol Biol. 370:459-470.
Frederiksen K, Rosenquist H, Jergensen K, Wilcks A. 2006. Occurrence of natural Bacillus thuringiensis contaminants and residues Bacillus thuringiensis-based insecticides on fresh ffruits and vegetables. Appl Environ Microbiol. 72:3435-3440.
Gao Q, Smith JC, Tsopmo A. 2014. Optimized protamex digested oat brand proteins: Antioxidant properties and identification of new peptides. Austin J Nutri Food Sci. 2:1-6.
Grau-Campistany A, Strandberg E, Wadhwani P, Reinchert J, Burk J, Rabana F, Ulrich AS. 2015. Hidrophobic mismatch demonstrated for membranolytic peptides, and their use as molecular rulers to measure bilayer thickness in native cells. Sci Rep. 5:9388.
Gutierrez-Chavez AJ, Martinez-Ortega EA, Valencia-Posadas M, Leon-Galvan MF, de la Fuente-Salcido NM, Bideshi DK, Barboza-Corona JE. 2015. Potential use of Bacillus thuringiensis bacteriocin to control antibiotic-resistant bacteria associated with mastitis in dairy goat. Folia Microbiol [Epub ahead of print].
Hayes M, Ross RP, Fitzgerald GF, Hill C, Stanton C. 2006. Ceasein-derived antimicrobial peptides generated by Lactobacillus acidophilus DPC6026. Appl Environ Microbiol. 72:2260-2264.
Henriksen JR, Etzerodt T, Gjetting T, Andresen TL. 2014. Side chain hydrophobicity modulates therapeutic activity and membrane selectivity of antimicrobial peptide mastoparan-X. PLoS ONE. 9:e91007.
Huang Y, He L, Li G, Zhai N, Jiang H, Chen Y. 2014. Role of helicity of α-helical antimicrobial peptides to improve specificity. Protein Cell. 5:631-642.
Huang YH, Huang JH, Chen Y. 2010. Alpha-helical cationic antimicrobial peptides: relationships of structure and function. Protein Cell. 1:143-152.
Jiang Z, Vasil AI, Vasil ML, Hodges RS. 2014. “Specificity determinants†improve therapeutic indices of two antimicrobial peptides piscidin 1 and dermaseptin S4 againts the Gram-negative pathogens Acinetobacter baumannii and Pseudomonas aeruginosa. Pharmaceuticals. 7:366-391.
Juretic D, Vukicevic D, Ilic N, Antcheva N, Tossi A. 2009. Computational design of high selective antimicrobial peptides. J Chem Inf Model. 49:2873-2882.
Keepers TR, Gomez M, Celeri C, Nichols WW, Krause KM. 2014. Bactericidal activity, absence of serum effect and time-kill kinetics of Ceftazidime-Avibactam against β-lactamase-producing Enterobacteriaceae and Pseudomonas aeruginosa. Antimicrob Agents Chemother. 58:5297-5305.
Kent RM, Guinane CM, O’Connor PM, Fitzgerald GF, Hill C, Stanton C, Ross RP. 2012. Production of the antimicrobial peptides Caseicin A and B by Bacillus isolates growing on sodium caseinate. Lett Appl Microbiol. 55:141-148.
Kim S, Kim SS, Lee BJ. 2005. Correlation between the activities of alpha-helical antimicrobial peptides and hydrophobicities represented as RP HPLC retention time. Peptides, 2050-2056.
Kweon C, Choi S, Kwon H, Kim E, Kang H, Moon J, Jang G, Lee H, Kang S, Kim J. 2012. Isolation, characterization, and evaluation of Bacillus thuringiensis isolated from cow milk. Korean J Vet Res. 52:169-176.
Laverty G, Gorman SP, Gilmore BF. 2011. The potential of antimicrobial peptides as biocides. Int J Mol Sci. 12:6566-6596.
Lazaridis T, He Y, Prieto L. 2013. Membrane interaction and pore formation by the antimicrobial peptide protegrin. Biophys J. 104:633-642.
Lorenzon EN, Cespedes GF, Vicente EF, Nogueira LG, Bauab TM, Castro MS, Cilli EM. 2012. Effect of dimerization on the structure and biological activity of antibacterial peptide Ctx-a. Antimicrob Agents Chemother 56, 3004-3010.
Mai XT, Huang J, Tan J, Huang Y, Chen Y. 2015. Effects and mechanisms of secondary structure on the antimicrobial activity and specificity of antimicrobial peptides. J Pept Sci. 7:561-568.
Maupetit J, Derreumaux P, Tuffery P. 2009. PEP-FOLD: an online resource for de novo peptide structure prediction. Nucleic Acids Res. 37: W498-503.
Maupetit J, Derreumaux P, Tuffery P. 2010. A fast and accurate method for large-scale de novo peptide structure prediction. J Comput Chem. 31:726-738.
Monteiro S, Carreira A, Freitas R, Pinheiro AM, Ferreira RB. 2015. A nontoxic polypeptide oligomer with fungicide potency under agricultural conditions which is equal or grater than that of their chemical counterpart. PLoS ONE. 10:e122095.
Nguyen LT, de Boer L, Zaat SAJ, Vogel HJ. 2011. Investigating the cationic side chains of the antimicrobial peptide tritrpticin: Hydrogen bonding properties govern its membrane-disruptive activities. Biochim Biophys Acta. 1808:2297-2303.
Nielsen SL, Frimodt-Moller N, Kragelund BB, Hansen PR. 2007. Structure-activity study of the antibacterial peptide fallaxin. Protein Sci. 16:1969-1976.
Potocnik K, Gantner V, Kuterova K, Cividini A. 2011. Mare’s milk: Composition and protein fraction in comparison with different milk species. Mljekarstvo. 61:107-113.
Prior RL, Wu X, Schaich K. 2005. Standardized methods for determination of antioxidant capacity and phenolics in foods and dietary supplements. J Agric Food Chem. 53:4290-4302.
Rahnamaeian M, Cytrynska M, Zdybicka-Barabas A, Dobslaff K, Wiesner J, Twyman RM, Zuchner T, Sadd BM, Regoes RR, Schimid-Hempel P, Vilcinskas A. 2015. Insect antimicrobial peptides show potentiating functional interactions againts Gram-negative bacteria. Proc R Soc B. 282:1-10.
Rath A, Glibowicka M, Nadeu VG, Chen G, Deber CM. 2009. Detergent binding explains anomalous SDS-PAGE migration of membrane proteins. PNAS. 106:1760-1765.
Rosenfeld Y, Lev N, Shai Y. 2010. Effect of the hydrophobicity to net ositive charge ratio on antibacterial and anti-endotoxin activities of structurally similar antimicrobial peptide. Biochemistry. 49:853-861.
Saravan R, Bhattacharjya S. 2011. Oligomeric structure of chatelicidin antimicrobial peptide in dodecylphosphocholine micelle determined by NMR spectroscopy. Biochimica et Biophysica Acta. 1808:369-381.
Shen Y, Maupetit J, Derreumaux P, Tuffery P. 2014. Improved PEP-FOLD approach for peptide and miniprotein structure prediction. Int Chem Theor Comput. 10:4745-4758.
Thaipong K, Boonprakob U, Crosby K, Cisneros-Zevallos L, Byrne DH. 2006. Comparison of ABTS, DPPH, FRAP, and ORAC assays for estimation antioxidant activity from guava fruit extracts. J Food Comp Anal. 19:669-67.
Thevenet P, Shen Y, Maupetit J, Guyon F, Derreumaux P, Tuffery P. 2012. PEP-FOLD: an updated de novo structure prediction server for both linear and disulfide bonded cyclic peptides. Nucleic Acids Res. 40:W288-293.
Uniacke-Lowe T, Huppertz T, Fox P. 2010. Equine milk protein: Chemistry, structure and nutritional significance. Int Dairy J. 20:609-629.
Vermeer LS, Abbate V, Ruh E, Bui TT, Wilkinson LJ, Kanno T, Jumagulova E, Kozlowska J, Patel J, McIntyre CA, Yam WC, Siu G, Atkinson AA, Lam JKW, Bansal SS, Drake AF, Mitchell GH, Mason AJ. 2012. Conformational flexibility determines selectivity and antibacterial, antiplasmodial, and anticancer potency of cationic α-helical peptides. J Biol Chem. 287:34120-34133.
Wilcks A, Smidt L, Bahl MI, Hansen BM, Andrup L, Hendriksen NB, Licht TR. 2008. Germination and conjugation of Bacillus thuringiensis subsp. israelensis in the intestine of gnotobiotic rats. J Appl Microbiol. 104:1252-1259.
Zhang Z, Hao H, Tang Z, Zou Z, Zhang K, Xie Z, Babe L, Goedegebuur F, Gu X. 2015. Identification and caharacterization of a new alkaline thermolysine-like protease, BtsLP1, from Bacillus thuringiensis serovar Sichuansis strain MC28. J Microbiol Biotechnol. 25:1281-1290.
Zhou G, Liu H, Yuan Y, Yuan Z. 2008. The occurrence of Bacillus cereus, B. thuringiensis and B. mycoides in Chinese pasteurized full fat milk. Int J Food Microbiol. 121:1995-2000.
Refbacks
- There are currently no refbacks.
This work is licensed under a Creative Commons Attribution 4.0 International License.
