The Change Rules On Muscle Protein Of Litopenaeus Vannamei Induced T-2Toxin

Feeds were seriously polluted by T-2toxin. T-2toxin can inhibit the synthesis of DNAand RNA, and that inhibit the synthesis of protein by interrupt translation to start. With theprocess of T-2toxin accumulation in the shrimp, muscle quality of shrimp changed.The object of research is Litopenaeus vannamei. The feeds has been artificialcontamination of T-2toxin, which was different set of exposure dose(0、1.2、2.4、4.8、12.2mg/kg·feed). Shrimp muscle protein was extraction and randomly divided into exposuregroup and control group, and that measured the content by Micro-Kjeldahl and Coomassiebrilliant blue method. Changes of soluble protein were submitted to analysis throughSDS-PAGE electrophoresis. Some targets, such as ATPase activity, sulfhydryl content andATP sensitivity, have been analysis by the conventional physical and chemical method. Theresults showed that the effects of T-2toxin were significantly（p<0.05）on muscle proteinclassification component. High dose T-2toxin induced actomyosin degradation. Acorrelation was found between the gray scale of42kD band in soluble protein and T-2toxin exposure dose, which is expected as a potential indicator to evaluate muscle qualityof white shrimp with T-2toxin induced. T-2toxin has significantly effect on differentprotein properties. The activity of Mg2+-ATPase and Ca2+-Mg2+-ATPase gradually reducedas the increasing T-2toxin exposure dose. Therefore, the Mg2+target enzyme can be usedas a marker to evaluate the changes of Litopenaeus vannamei muscle protein qualityinduced by T-2toxin.We established two-dimensional electrophoresis separation techniques for shrimpmuscle protein, and optimized the separation conditions. The results show that there arefive optimal parameters for successfully obtaining2-DE spectrum with high separatingdegree and resolving capability. First, the way of extraction is nitrogen grinding andultrasonic broken; Second, lysate is chiefly made of7M urea and2M thiourea, which canextract the shrimp muscle holoprotein; Third, using7cm, pH4-7IPG strips; Forth, loadingquantity of sample is180μg, sample loading method is passive; Last, concentrations ofseparating gel is12.5%in the course of2to electrophoresis, traditional method iscoomassie R-250dyeing method. This article emphasized to further explore the changerule about shrimp muscle protein induced by T-2toxin, and laid the foundation for filtratemarker protein.Shrimp muscle protein with different T-2toxin dose groups was separated using two-dimensional electrophoresis separation technology. The result is that we get30differentially expressed protein spots, and there are12protein spots among themconcerned with T-2toxin concentration. They were analyzed to peptide mass fingerprintingand MS/MS by MALDI-TOF/TOF. Based on MASCOT, there were11protein spots and1unknown protein spot. Homologous protein of11protein spots are SUMO-1, argininekinase, triosephosphate isomerase, arrestin112, Arginine kinase B chain,70kD heat shockprotein (partial) and vitellogenin. The general trend of protein spots expression quantity israised first and cut after in five T-2toxin dose groups. No.2288point is Arginine Kinase,its expression quantity went up under low T-2toxin dose, and then went disappear. Itmeans that Arginine Kinase has high sensitive to high T-2toxin dose. Therefore, ArginineKinase is labelled protein in this paper.Above all, T-2toxin has significant effects on prawn muscle protein. The change trendof muscle protein presents declining while T-2toxin exposure dose increased. It is the mainreason that shrimp muscle quality declined because of T-2toxin. This study provides aspecific parameter and theoretical basis that T-2toxin induces vannamei muscle qualitychange. It also provides a scientific basis for green product quality of Litopenaeusvannamei.