| The test used different induce method on Tenebrio molito.Adopt 6 induce methods:the ultraviolet rays revulsion,the ultrasonic wave revulsion,the needing revulsion,the bacterium fluid soak revulsion,starve revulsion,and bacterium fluid of inject revulsion,act.Per method adopted different dosage or induce time and cultured different time after induction,then tested by bacteriostasis in order to decided that if the antibacterial active compound producted or not.To observe the antibacterial activity strong or weak when their were created.The result was that using the bacterium fluid of injects revulsion and injects 1μl/head then cultured 48h after induction was the best method of induction.Next was the bacterium fluid soak revulsion and starve revulsion.In addition method of others even if had unvarying bacteriostasis effect but quiet.In the test, different method of induction were adopted various cultivate time after induction.For example:24h,48h and 72h.As a result,bacteriostasis activity of blood-lymph also exist tremendous disparation if cultured different time after induction.Cultivate 48h was optimization.If less than 48h,polypide cannot emerge effective defence mechanism therefore bacteriostasis activity was not ideal.If more than 48h,bacterium fluid would be persistence for a long time in the polypide.Furthermore multiply speed of bacterium was quickly.So that the speed of emerging defence system was slower than bacterium growing in the polypide,to result in the polypide was unable to pick oneself up,motoricity drop out,extraction quantity of blood-lymph diminished, even caused Tenebrio molitor death.It was good for observation bacteriostasis activity neither.This text had compared detail on many kinds of methods of induction,then screened the best method of induction by first time.The test extracted blood-lymph by the best method of induction,in order to survey and evaluate biologic activity of antibacterial peptide.In the test,it determined heat stability,stability of freeze thawing,prolease stability,effect of antibacterial peptide activity by organic solvent and Ph value.Finally eventuate was that antibacterial peptide from Tenbrio molitor had better heat stability.Disposal by 10min also had better effective of bacteriostasis between 90—100℃.After freeze thawing again and again,the diameter of bacteriostasis circle had invariably grown down,but the difference by freeze thawing less than 4times was not significant. When freeze thawing 6—10times,bacteriostasis activity reached significance level which compared to check, but it can observe the result of bacteriostasis still to remain.Bacteriostasis rate had broken down a little after the action of typsinase and pepsase,but prolease K had little influence on bacteriostasis activity at all. Although bacteriostasis activity had broken down a little when Ph value at 5.0—9.0,but it can not cause antibacterial activity deprivation.If Ph value less than 5.0 or more than 9.0,its bacteriostasis activity would be suffered tremendous influence,probably appear no activity.Methanol and n-butanol were without significant influence on bacteriostasis activity of antibacterial peptide.But bacteriostasis activity of antibacterial peptide particular had been effect by alcohol,chloroform and isopropand.The finally conclusion was that antibacterial peptide from Tenebrio molitor had better heat stability,freeze thawing stability.After affect by prolease bacteriostasis activity was breakdown to some extent.In addition,bacteriaostasis activity had been effect by Ph value or organic solvent more or less.The antibacterial peptides were isolated and purified from hemolymph of Tenebrio molitor larvae through Sephadex chromatography.Their antibacterial activity was detected on agarose plates.After Sephadex G50 filtration,an antibacterial fraction was obtained,which showed potent antibacterial activity on E.coli.To obtain 2 visible summit of activity,the result demonstrated that from 5 pipe to 19 pipe all had bacteripstasis activity.That explained the assemblage mixing by multiple composition,as a result,the separated was emphatically not thorough.Then let the part which had compt put together to condense.After Sephadex G25 filtration,an antibacterial fraction was obtained,which showed potent antibacterial activity on E.coli.The antibacterial fraction was as composed of many antibacterial peptides.About how to obtain the monomer,it should be study and research further.This study was the foundation of the further identifying and purification to antibacterial peptides from Tenebrio molitor.And in detail compared and selected the induction that how to make Tenebrio molitor to produced antibacterial peptide first,then determined a more excellent and reasonable induction.By researching and anglicizing the biological characteristics to provided theoretical foundations to preservation and using the antibacterial peptide of Tenebrio molitor.
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