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Protein & Peptide Letters

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ISSN (Print): 0929-8665
ISSN (Online): 1875-5305

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Research Article

EGLLGDVF: A Novel Peptide from Green Mussel Perna viridis Foot Exerts Stability and Anti-inflammatory Effects on LPS-Stimulated RAW264.7 Cells

Author(s): Ila Joshi and Rasool Abdul Nazeer*

Volume 27, Issue 9, 2020

Page: [851 - 859] Pages: 9

DOI: 10.2174/0929866527666200224111832

Price: $65

Abstract

Background: Green mussel Perna viridis is a bivalve mollusc which is native to the Indian coast and can be found in the Indo-Pacific as well as Asia-Pacific regions. This study evaluates the P. viridis foot (PVF) as a source of an anti-inflammatory peptide.

Objective: To characterize and evaluate the possibility of pro-inflammatory cytokines, nitric oxide (NO) as well as cyclooxygenase (COX)-2 reduction in RAW264.7 cells and to analyze functional aspects of the derived peptide from PVF.

Materials and Methods: The PVF was hydrolysed with different enzymes and the antiinflammatory activity of hour hydrolysates were evaluated using HRBC Membrane Stabilization (HMS) against hypotonicity induced haemolysis and Albumin Denaturation (AD) inhibition from induced heat assays. Later, the active hour hydrolysate was separated by ultrafiltration and purified using Size-Exclusion Chromatography (SEC). Further, the purified peptide’s sequence was identified using LC-MS/MS and functional properties were determined. Also, the peptide was observed for its anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated RAW264.7 cells for pro-inflammatory cytokines, NO production and COX-2 activation.

Results: Among the four enzymes 6th hour alcalase hydrolysate exhibited potent anti-inflammatory activity and was sequentially fractioned with molecular weight cut-offs; further active fraction (30- 10 kDa) was purified. The active peak-II was identified as EGLLGDVF (849.435 Da) and exhibited decent functional aspects. The peptide successfully reduced the production of pro-inflammatory cytokines, NO and COX-2 activation; and down-regulated the iNOS and COX-2 protein expression in LPS-stimulated RAW264.7 cells.

Conclusion: Our study indicates that EGLLGDVF derived from PVF has potential antiinflammatory applications applicable in food and pharmaceutical industries.

Keywords: P. viridis foot, anti-inflammatory peptide, enzymatic hydrolysis, mass spectrometry, functional property, RAW264.7 cells.

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[1]
Anosike, C.A.; Obidoa, O.; Ezeanyika, L.U. Membrane stabilization as a mechanism of the anti-inflammatory activity of methanol extract of garden egg (Solanum aethiopicum). Daru, 2012, 20(1), 76.
[http://dx.doi.org/10.1186/2008-2231-20-76 ] [PMID: 23351977]
[2]
Ackerman, N.R.; Beebe, J.R. Release of lysosomal enzymes by alveolar mononuclear cells. Nature, 1974, 247(5441), 475-477.
[http://dx.doi.org/10.1038/247475a0 ] [PMID: 4818545]
[3]
Opie, E.L. On the relation of necrosis and inflammation to denaturation of proteins. J. Exp. Med., 1962, 115, 597-608.
[http://dx.doi.org/10.1084/jem.115.3.597 ] [PMID: 14482110]
[4]
Chakraborty, K.; Chakkalakal, S.J.; Joseph, D. Response of pro-inflammatory prostaglandin contents in anti-inflammatory supplements from green mussel Perna viridis L. in a time-dependent accelerated shelf-life study. J. Funct. Foods, 2014, 7, 527-540.
[http://dx.doi.org/10.1016/j.jff.2014.01.003]
[5]
Sreejamole, K.L.; Radhakrishnan, C.K.; Padikkala, J. Anti-inflammatory activities of aqueous/ethanol and methanol extracts of Perna viridis Linn. in mice. Inflammopharmacology, 2011, 19(6), 335-341.
[http://dx.doi.org/10.1007/s10787-010-0075-z ] [PMID: 21359864]
[6]
Lowry, O.H.; Rosebrough, N.J.; Farr, A.L.; Randall, R.J. Protein measurement with the Folin phenol reagent. J. Biol. Chem., 1951, 193(1), 265-275.
[PMID: 14907713]
[7]
Lee, S.H.; Chang, D.W.; Lee, B.J.; Jeon, Y.J. Antioxidant activity of solubilized Tetrasel missuecica and Chlorella ellipsoidea by enzymatic digests. J. Food Sci. Nutr., 2009, 14, 21-28.
[8]
Wanasundara, P.K.J.P.D.; Ross, A.R.; Amarowicz, R.; Ambrose, S.J.; Pegg, R.B.; Shand, P.J. Peptides with angiotensin I-converting enzyme (ACE) inhibitory activity from defibrinated, hydrolyzed bovine plasma. J. Agric. Food Chem., 2002, 50(24), 6981-6988.
[http://dx.doi.org/10.1021/jf025592e ] [PMID: 12428947]
[9]
Mizushima, Y. Screening test for anti-rheumatic drugs. Lancet, 1966, 2, 443-448.
[http://dx.doi.org/10.1016/S0140-6736(66)92756-5]
[10]
Kulkarni, A.; Govindappa, M.; Chandrappa, C.; Ramachandra, Y.; Koka, P.S. Phytochemical analysis of Cassia fistula and it’s in vitro antimicrobial, antioxidant and anti-inflammatory activities. Adv. Med. Plant Res., 2015, 3, 8-17.
[11]
Pearce, K.N.; Kinsella, J.E. Emulsifying properties of proteins: Evaluation of a turbidimetric technique. J. Agric. Food Chem., 1978, 26, 716-723.
[http://dx.doi.org/10.1021/jf60217a041]
[12]
Sathe, S.K.; Salunkhe, D.K. Functional properties of the Great Northern bean (Phaseolus vulgaris L.) proteins: Emulsion, foaming, viscosity and gelation properties. J. Food Sci., 1981, 46, 71-74.
[http://dx.doi.org/10.1111/j.1365-2621.1981.tb14533.x]
[13]
Mosmann, T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J. Immunol. Methods, 1983, 65(1-2), 55-63.
[http://dx.doi.org/10.1016/0022-1759(83)90303-4 ] [PMID: 6606682]
[14]
Sun, J.; Zhang, X.; Broderick, M.; Fein, H. Measurement of nitric oxide production in biological systems by using griess reaction assay. Sensors (Basel), 2003, 3, 276-284.
[http://dx.doi.org/10.3390/s30800276]
[15]
Arihara, K.; Ohata, M. Functional properties of bioactive peptides derived from meat proteins. In: Advanced Technologies for Meat Processing; Toldra, F., Ed.; Springer: New York, NY, USA, 2006; pp. 245-274.
[http://dx.doi.org/10.1201/9781420017311.ch10]
[16]
Hernandez-Ledesma, B.; Quiros, A.; Amigo, L.; Recio, I. Identification of bioactive peptides after digestion of human milk and infant formula with pepsin and pancreatin. Int. Dairy J., 2007, 17, 42-49.
[http://dx.doi.org/10.1016/j.idairyj.2005.12.012]
[17]
Lueangsakulthai, J.; Phosri, S.; Theansungnoen, T.; Jangpromma, N.; Temsiripong, T.; Mckendrick, J.E.; Khunkitti, W.; Klaynongsruang, S. Novel antioxidant and anti-inflammatory peptides from the Siamese crocodile (Crocodylus siamensis) hemoglobin hydrolysate. Biotechnol. Appl. Biochem., 2018, 65(3), 455-466.
[http://dx.doi.org/10.1002/bab.1628 ] [PMID: 29164679]
[18]
Halliwell, B.; Whiteman, M. Measuring reactive species and oxidative damage in vivo and in cell culture: How should you do it and what do the results mean? Br. J. Pharmacol., 2004, 142(2), 231-255.
[http://dx.doi.org/10.1038/sj.bjp.0705776 ] [PMID: 15155533]
[19]
dBagad, Y.M.; Umarkar, A.R.; Tatiya, A.U.; Surana, S.J. Investigation of anti-inflammatory and analgesic activity of Bridelia airyshawii (Euphorbiaceae). J. Pharm. Res., 2011, 4, 1326-132.
[20]
Hasegawa, S.; Ichiyama, T.; Sonaka, I.; Ohsaki, A.; Okada, S.; Wakiguchi, H.; Kudo, K.; Kittaka, S.; Hara, M.; Furukawa, S. Cysteine, histidine and glycine exhibit anti-inflammatory effects in human coronary arterial endothelial cells. Clin. Exp. Immunol., 2012, 167(2), 269-274.
[http://dx.doi.org/10.1111/j.1365-2249.2011.04519.x ] [PMID: 22236003]
[21]
Li, X.; Bradford, B.U.; Wheeler, M.D.; Stimpson, S.A.; Pink, H.M.; Brodie, T.A.; Schwab, J.H.; Thurman, R.G. Dietary glycine prevents peptidoglycan polysaccharide-induced reactive arthritis in the rat: Role for glycine-gated chloride channel. Infect. Immun., 2001, 69(9), 5883-5891.
[http://dx.doi.org/10.1128/IAI.69.9.5883-5891.2001 ] [PMID: 11500467]
[22]
Kristinsson, H.G.; Rasco, B.A. Fish protein hydrolysates: Production, biochemical, and functional properties. Crit. Rev. Food Sci. Nutr., 2000, 40(1), 43-81.
[http://dx.doi.org/10.1080/10408690091189266 ] [PMID: 10674201]
[23]
Kumar, N.S.S.; Nazeer, R.A.; Ganesh, R.J. Functional properties of protein hydrolysates from different body parts of horse mackerel (Magalaspis cordyla) and croaker (Otolithes ruber). Med. J. Nutrition Metab., 2012, 5, 105-110.
[http://dx.doi.org/10.1007/s12349-011-0078-3]
[24]
Taheri, A.; Anvar, S.A.A.; Ahari, H.; Fogliano, V. Comparison the functional properties of protein hydrolysates from poultry by-products and rainbow trout (Onchorhynchus mykiss) viscera. Iran. J. Fish. Sci., 2013, 12, 154-169.
[25]
dos Santos, S.A.; Martins, V.G.; Sallas-Mellado, M.; Prentice, C. Evaluation of functional properties in protein hydrolysates from blue wing sea robin (Prionotus punctatus) obtained with different microbial enzymes. Food Bioprocess Technol., 2011, 4, 1399-1406.
[http://dx.doi.org/10.1007/s11947-009-0301-0]
[26]
Lam, R.S.; Nickerson, M.T. Food proteins: A review on their emulsifying properties using a structure-function approach. Food Chem., 2013, 141(2), 975-984.
[http://dx.doi.org/10.1016/j.foodchem.2013.04.038 ] [PMID: 23790876]
[27]
Adebowale, K.; Lawal, O. Comparative study of the functional properties of bambarra groundnut (Voandzeia subterranean), jack bean (Canavalia ensiformis) and mucuna bean (Mucuna pruriens) flours. Food Res. Int., 2004, 37, 355-365.
[http://dx.doi.org/10.1016/j.foodres.2004.01.009]
[28]
Solini, A.; Muscelli, E.; Stignani, M.; Melchiorri, L.; Santini, E.; Rossi, C.; Astiarraga, B.D.; Rizzo, R.; Baricordi, O.R. Soluble human leukocyte antigen-g expression and glucose tolerance in subjects with different degrees of adiposity. J. Clin. Endocrinol. Metab., 2010, 95(7), 3342-3346.
[http://dx.doi.org/10.1210/jc.2009-2747 ] [PMID: 20427497]
[29]
Li, D.; Chen, J.; Ye, J.; Zhai, X.; Song, J.; Jiang, C.; Wang, J.; Zhang, H.; Jia, X.; Zhu, F. Anti-inflammatory effect of the six compounds isolated from Nauclea officinalis Pierrc ex Pitard, and molecular mechanism of strictosamide via suppressing the NF-κB and MAPK signaling pathway in LPS-induced RAW 264.7 macrophages. J. Ethnopharmacol., 2017, 196, 66-74.
[http://dx.doi.org/10.1016/j.jep.2016.12.007 ] [PMID: 27989509]
[30]
Ahn, C.B.; Cho, Y.S.; Je, J.Y. Purification and anti-inflammatory action of tripeptide from salmon pectoral fin byproduct protein hydrolysate. Food Chem., 2015, 168, 151-156.
[http://dx.doi.org/10.1016/j.foodchem.2014.05.112 ] [PMID: 25172694]
[31]
Hwang, D.; Kang, M.J.; Jo, M.J.; Seo, Y.B.; Park, N.G.; Kim, G.D. Anti-inflammatory activity of β-thymosin peptide derived from pacific oyster (Crassostrea gigas) on NO and PGE2 production by down-regulating NF-κB in LPS-induced RAW264.7 macrophage cells. Mar. Drugs, 2019, 129, 1-11.
[http://dx.doi.org/10.3390/md17020129]

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