Separation, Purification Monopterus Albus Antimicrobial Peptide, The Preliminary Study Of The Crude Antimicrobial Peptide Protecting Monopterus Albus From A.hydrophila Challenge

Antimicrobial peptides(AMPs)are a recently discovered type of antimicrobial activity peptide and play an important role in the innate host defense mechanisms of most living organisms.But related researches on aquatic animal have few reported before.Now research of Monopterus albus AMPs hasn't reported besides our laboratory.Three experiments were studied the antibacterial activity of the AMPs crude sample from Monopterus albus's skin,skin mucus,then were purification AMPs of the skin mucus,the effect of crude antibacterial peptide from Monopterus albus skin mucus Protecting mice from lethal dosage 50%(LD₅₀)A.hydrophila challenge were observed.Experiment 1.antibacterial peptides Raw Extraction of differ tissue from Monopterus albusThis experiment was used methods of Tris-HCl extraction and acetic acid extraction on antibacterial peptide crude sample from induced Monopterus albus's skin mucus,blood,skin,liver,spleen,intestine,kidney.The methods was succeed in separating antibacterial peptide crude sample of skin mucus,skin,intestine.This showed that the antibacterial activity of the raw sample from skin mucus,skin and intestine can be detected inhibition zone by A.hydrophila and salmonelladerby,but the raw sample from blood,liver,spleen,kidney can not.Experiment 2.The crude extract protecting Monopterus albus from A.hydrophila challengeThe effect of crude peptide from Monopterus albus skin mucus(160mg/mL,80mg/mL,40 mg/mL)protecting Monopterus albus from LD₅₀A.hydrophila(1×10¹⁰CFU/kg)challenge were observed.(160mg/mL)could survival all Monopterus albus from LD₅₀of A.hydrophila challenge,crude peptide from Monopterus albus skin mucus also protected Monopterus albus from tissue damage by LD₅₀of A.hydrophila challenge.The experiment had prompted:crude peptide from Monopterus albus skin mucuscould protecting Monopterus albus from LD₅₀A.hydrophila(1×10¹⁰CFU/kg)challenge.Experiment 3.Separating and purification of antibacterial peptide from Monopterus albus skin mucusDifferent protein components were separated by using Sephadex G50 gel filtration sequencely,last purification antibacterial peptides was completed by Macro-Prep High S in this experiment.Three ultraviolet absorption peak of antibacterial peptide were collected in Sephadex G50 filtration chromatography,and rough screening of antibacterial activity was investigated.The antibacterial activity was detected from peak 3.The antibacterial samples that collected by Sephadex G50 were separated by Macro-Prep High S.Two ultraviolet absorption peak of antibacterial peptide were collected in Macro-Prep High S.rough screening of antibacterial activity was investigated.The antibacterial activity was detected from peak 2.After SDS-PAGE,the Monopterus albus antibacterial peptide showed a single protein band,and the molecular weight was about 8400Da.