https://doi.org/10.17113/ftb.57.02.19.5964       


Nitric Oxide Synthesis Inhibition and Anti-Inflammatory Effect of Polypeptide Isolated from Chicken Feather Meal in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages   


Aurachorn Inkanuwat¹, Romteera Sukaboon², Onrapak Reamtong³, Pravit Asawanonda⁴, Ake Pattaratanakun⁵, Tanatorn Saisavoey⁶, Papassara Sangtanoo6 and Aphichart Karnchanatat⁶*  


¹Technopreneurship and Innovation Management Program, Graduate School, Chulalongkorn University, 254 Phayathai Road, Pathumwan, Bangkok 10330, Thailand 
²Program in Biotechnology, Faculty of Science, Chulalongkorn University, 254 Phayathai Road, Pathumwan, Bangkok 10330, Thailand 
³Department of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine, Mahidol University, 420/6 Ratchawithi Road, Ratchathewi, Bangkok 10400, Thailand 
⁴Department of Medicine, Faculty of Medicine, Chulalongkorn University, 254 Phayathai Road, Pathumwan, Bangkok 10330, Thailand 
⁵Department of Marketing, Faculty of Commerce and Accountancy, Chulalongkorn University, 254 Phayathai Road, Pathumwan, Bangkok 10330, Thailand 
⁶Research Unit in Bioconversion/Bioseparation for Value-Added Chemical Production, Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, 254 Phayathai Road, Pathumwan, Bangkok 10330, Thailand   



Article history: 
Received: 2 August 2018  
Accepted: 26 March 2019 
  



Key words: 
nitric oxide, anti-inflammatory activity, chicken feather meal, protein hydrolysate, polypeptide, macrophage RAW 264.7  



Summary: 
Nitric oxide (NO) plays a key role in the pathogenesis of inflammation and has been implicated in endotoxin-induced tissue injury. Chicken feather meal is a rich source of amino acids that may serve as a peptide hydrolysate to inhibit NO activity. Anti-inflammatory hydrolysates of chicken feather meal were prepared and fractionated into five samples based on molecular mass. The smallest fraction (<0.65 kDa) exhibited the highest NO inhibitory activity without cytotoxicity towards macrophage RAW 264.7 cells. Further subfractions were sufficient to obtain amino acid sequences by Q-TOF LC-MS/MS ESI analysis. Of these, the SNPSVAGVR (885.97 Da) peptide and its corresponding pure synthetic peptide have inhibitory activity against NO production by RAW 264.7 cells (IC₅₀=(55.2±0.2) mM) without cytotoxicity. Reverse transcriptase polymerase chain reaction (RT-PCR) and quantitative real-time RT-PCR results revealed that the peptide of the obtained fraction reduced transcript expression levels of the pro-inflammatory cytokines iNOS, TNF-α, COX-2 and IL-6 in lipopolysaccharide-stimulated RAW 264.7 cells. These results suggest that the peptides derived from the chicken feather meal protein could potentially be used as a promising ingredient in functional foods or nutraceuticals against inflammatory diseases.    




*Corresponding author:    +6622188078  
                                             +6622188069
                                              Aphichart.K@chula.ac.th