PSP Binding Protein:It's Purification And Neutralization To Paralytic Shellfish Poisoning(PSP) Toxicity

Paralytic shellfish poisoning(PSP)is a kind of marine biotoxin,which can bolck sodium channels in the nerve cell and cause paralysis of human nerve system.PSP toxins can lead to poisoning death of humanity after ate the tainted shellfish.But for now there is no specific antidote to against PSP.Based on previous research,taking shore crab Thalamita crenata inhabited near Zhanjiang leizhou peninsula as the research objects,analyze and compare three purifying methods of PSP binding protein(PSPBP),further expound its physical and chemical properties,stablityand neutralization effect to PSP.The conclusion will provide scientific theoretical basis for the PSP and the research of antidotes.This article mainly studied from the following several aspects.1.We get the best purification method of the PSPBP by comparing the ammonium sulfate precipitation,gel column chromatography and affinity chromatography.These results indicate that Using the affinity resin AF-Epoxy-650 M to purify PSPBP can attain the best purifying effect.Although the proteins were not a single protein,they also have good performance to neutralize the toxicity of PSP.2.The physicochemical properties and stablity of affinity proteins isolated early were stutied with HPLC.Results showed that the best effect of combination may be got when the reaction p H 7.15,the reaction temperature 4 °C and the reaction time 1 h.Divalent metal ions of Mg Cl2,Ca Cl2 had significant inhibition effect on the combination between affinity proteins and PSP.Temperature was the main factor to influence the stablity of affinity protein.The higher the temperature was,the activity of affinity protein was more difficult to remain.As the storing temperature was 4 °C,the toxin binding rate reached the highest.Different denaturants had various functions on destroying the activity of activity of affinity,1% SDS had the most effect followed by 5% DTT and 3 M guanidine hydrochloride.3.The effcet of the affinity protein to neutralize PSP were studied with AOAC mouse bioassay and HPLC.When combined PSP with different concentrations of PSPBP.The PSP binding rate increased before decreasing while the concentration of protein rose,withthe increase of the concentration of protein.The presence of high concentrations of PSP biding protein greatly extended the time of death in mouse.