Study On The Antibacterial Properties Of The Body Cavity Fluid Of Phasolosma Esculenta

In an background which antibiotics are abused and cause serious harm to people,it is particularly important to find new natural antibacterial active substances.Recent years,natural antibacterial active substances extracted from invertebrate body cavity fluid have also been became popular.As a unique aquatic product resource in China,the Phasolosma esculenta has sufficient body cavity fluid content.However,the lack of development and utilization has caused a great waste of body cavity fluid resources.In this paper,we studied the technique of acquiring body cavity fluids using the Phasolosma esculenta as raw materials,and analyzed the antibacterial properties of body cavity fluids using the bacterial growth curve method and the Oxford cup method.At the same time,dextran gel column chromatography and liquid phase were used.Chromatography-mass spectrometry was used to separate and purify antibacterial proteins in body cavity fluid and to identify the amino acid sequence.The main experimental results are as follows:(1)Research on the extraction technology of Phasolosma esculenta body fluid.In the low-voltage electric shock method,the effect of body cavity fluid obtained under 6V voltage shock is better.Under this condition,the polysaccharide and protein concentrations in the unit volume(30 mL)of body cavity fluid are: 0.163 ± 0.005 mg/mL and 0.295 ± 0.006 mg / mL,respectively;The concentration of polysaccharide and protein in the body cavity fluid under direct dissection method(30 mL)were: 0.186±0.004 mg/mL and 2.205±0.005 mg/mL,respectively.(2)Research on the antibacterial properties of delicious cystic cyst body fluid.The delicious cystic cyst body fluid has strong inhibitory ability against Escherichia coli,Shewanella spp.,And Vibrio parahaemolyticus,while it is resistant to Staphylococcus aureus,Micrococcus lyticus,Bacillus subtilis,Pseudomonas aeruginosa The effect is not obvious;the polysaccharide and protein in the body cavity fluid are separated and the antibacterial experiment results on the above three sensitive bacteria are found respectively.(3)Study on the stability of the antibacterial protein in the body cavity fluid of the Phasolosma esculenta.The antibacterial protein is not resistant to high temperature,sensitive to ultraviolet rays,acid and alkali,sensitive to proteinase k and chymotrypsin,and metal ions have a significant effect on its antibacterial effect(P<0.05);The respectively minimum inhibitory concentrations of E.coli Shewanella spp.And Vibrio parahaemolyticus were 36 ?g/mL,72 ?g/mL,and 8 ?g/mL.(4)Isolation,purification and sequence identification of the antibacterial protein from t he body cavity fluid of Phascolosoma esculenta.The antibacterial protein of the body cavit y fluid of Phasolosma esculenta is a B6CPA3 protein.The protein contains 16 peptides,t he sequence coverage reaches 99.2%,the molecular weight is about 13.629 kDa,and the n umber of amino acids in the theoretical sequence of the protein is 120.The amino acid se quence of the antibacterial protein is: VFYDDLDNQHK(valine-phenylalanine-tyrosine-aspart icacid-asparticacid-leucine-aspartic acid-asparagine-glutamine-histamine Acid-lysine).