梅花鹿鹿角盘蛋白的分离纯化及其对大肠杆菌抑菌机制的初步研究

AbstractAntler plate is the bone part left after the antler of sika deer or red deer was sawed off.Itwill fall off naturally when the antler grows out in the next spring.Because of its shape,it iscalled antler plate or deer tray.As early as in the Han Dynasty,one of the four major works ofChinese medicine,there is a record of antler disc in the Shennong Materia Medica Classic."TheSutra":its main evil sores,carbuncle,chase evil gas,leaving blood in the Yin.It can becondensed into antler glue through water,which has the effect of nourishing liver and kidney,adding essence and stopping bleeding.It is not as strong as antler,but it is better than antler.Theremaining bone residue is antler cream,which has the effect of warming kidney and helpingYang,astringency and hemostasis.Antler disc is widely used in the treatment of mastitis,mammary gland hyperplasia,gynecological inflammation,enteritis,gastritis,sore swelling,lumbar cold pain,etc.It also has the effect of strengthening tendon and bone,promoting bloodcirculation,removing blood stasis and so on.Therefore,the development and utilization of antlerdisc has a good prospect.Proteins,one of the main functional components of the antler dish,have also attracted extensive attention because they are the material basis of life and animportant component of body cells and tissues.Therefore,this project takes the protein as theentry point to search for the bioactive protein in the antler dish and conduct a preliminary studyon its mechanism of action,aiming to provide data support for the development and utilizationof the valuable animal medicine.This experiment adopts acid extraction and alcohol sink,Sephadex G-25 gel in addition tosalt and a series of methods for sika deer antler plate of total protein in addition to salt to taste,to Sephacryl antler plate of total protein in addition to salt product S-100 gel filtrationchromatography separation of the antimicrobial activity of protein components,including SDSpolyacrylamide gel electrophoresis (sds-page),mass spectrometry,high performance liquid ofantler plate of total protein and antibacterial protein identification and analysis;Next,Escherichia coli was used as the research object.The bacteriostatic effect of total protein andantibacterial protein was studied by double dilution method of microbroth,and the minimumbacteriostatic concentration was determined.On this basis,UV spectrophotometer,lightmicroscope,enzyme plate,fluorescence microscope,flow cytometry and scanning electronmicroscopy were used to determine the effects of bacterial agglutination,cell membranepermeability,cell wall integrity and induced oxidative stress of E.coli.The results showed that the total protein content of the antler plate after desalination was90.3%.The protein component S2 with a molecular weight of It <20.1 kDa was obtained by