| The plant protein hydrolysates include some active peptides, this active peptide could not only have some nutritional function ,but also could promote animal growth, enhance immunity ability, advance the somatic ability of antivirus and so on. The research use plant protein hydrolysates enzymed by endoproteinase to do biological valuable analysis, and use the hydrolysates to do in vivo experiment to explore the hydrolysates' function on fish development and its mechanism, to offer the theory evidence for the The plant protein hydrolysates' exploration and application.1 Selection of the Optimum Conditions of Enzymatic Hydrolysis of Soybean meal with different EnzymesThe optimum enzymatic conditions of papain , trypsin and As1.398 subtilisin which hydrolyze Soybean meal(SBM) were studied with the degree of hydrolysis(DH) through mono-factor analysis and orthogonal test( L9 (34) ).The result showed that the optimum enzymatic conditions of papain were PH 7.5, temperature 50℃, time 7 hours ,enzyme concentration 5 percent of Soybean meal(SBM) protein .In such conditions, the degree of hydrolysis of soybean meal(SBM) can go up to 3.67%.The optimum conditions of trypsin were PH 7.0, temperature 50℃.time 7 hours ,enzyme concentration 5 percent of Soybean meal(SBM) protein. In such conditions, the degree of hydrolysis of soybean meal can go up to 5.53%; The optimum conditions of Asl.398 subtilisin were PH 6.5, temperature 50℃,time 7 hours ,enzyme concentration 5 percent of Soybean meal(SBM) protein. In such conditions, the degree of hydrolysis of soybean meal can go up to 8.73%.The hydrolysis activity of As1.398 subtilisin is the highest of three enzyme.2 Ultrafiltratione and SDS-PAGE analysis of common feeds plant protein hydrolysatesSoybean meal, cottonseed meal, rapeseed meal and peanut meal were hydrolyzed with the papain, trypsin and As1.398 subtilisin. Enzymolysis products of them were analysed with the method of ultrafiltratione and SDS-PAGE electrophoresis. The result showed that: As1.398 subtilisin was the best proteinase among three enzyme which hydrolyze soybean meal. The higher hydrolysis degree(DH) of soybean meal is, the more small molecular weights of enzymatic products of the soybean meal is, when soybean meal DH is 5.68％, the molecular weights which are bigger than 30KD is 73.09％, from 10KD to 30KD is 2.75％, from 3KD to 10KD is 13.42％, the smaller than 3KD is 10.75％,when soybean meal DH is 8.73％, the bigger than 30KD is 44.87％, from 10Kd to 30Kd is 8.83％, from 3Kd to 10Kd is 14.63％, the smaller than 3KD is 31.67％,when soybean meal was not hydrolyzed, the bigger than 30KD is 94.12％, from 10KD to 30KD is 3.06％, from 3Kd to 10Kd is 1.86％, the smaller than 3KD is 0.97％. As the optional conditions of enzymatic protein hydrolysis were the same, the soybean meal hydrolyzed by As1.398 subtilisin was the highest degree of hydrolysis, the second is cottonseed meal, the third is peanut meal, the lowest is rapeseed meal, the molecular weights below 10KD of enzymatic products were 46.3％, 25.13％, 13.38％and 12.58％. The molecular weights of enzymatic products were measured by sodium dodecy sulfafce-polyactivity gel electrophoresis(SDS-PAGE) in this experiment.3 Effect of plant protein hydrolysates on the growth of the mouse embryo fibroblasts cellPapain, trypsin and As1.398subtilisin were used to degrade soybean meal, cottonseed meal, rapeseed meal and peanut meal separately, then used hydrolysates to study the effect on the culture of the mouse embryo fibroblasts cell. The result indicated that all plant protein hydrolysates could promote the growth of the mouse embryo fibroblasts cell, and the most effective hydrolysates was the cottonseed meal degraded by As1.398subtilisin, the speed of cell growth of that was improved than that of hydrolysates of the peanut meal, rapeseed meal and soybean meal.4 Effects of plant proteins hydrolysates on proliferation of intestinal microorganisms of fishEscherichia coli, Bacillus and Lactobacillus were isolated from intestines of healthy carps by plate cultivation. Effects of plant proteins and their hydrolysates on the growth of three microorganisms were studied by liquid cultivation in vitro. The result was that: cottonseed meal, soybean meal, peanut meal and the hydrolysates of cottonseed, soybean, rapeseed meal and peanut meal observably promoted the proliferation of Bacillus and Lactobacillus. and than the effect was significantly different(p＜0.05): all of four meals and their hydrolysates could promote the proliferation of Escherichia coli except hydrotysates of cottonseed meal., but had no significant effects(p＞0.05).5 Effect of hydrolysates on the growth and immune indices of Alloggnogeuetic Crucian CarpThe basal diet as the control group, four hydrolysates and fish meal were added in test groups teed for Alloggnogeuetic crucian, fish raised for 61 days, the results demonstrated that: the gained weight rates of all experiment groups were higher than the control and the feed coefficient was reduced, the highest weight gain which was 32.13％higher than control group was in Cottonseed meal hydrolysates group(P＜0.05), gained weight rate of others were 3.67％,5.83％,12.17％and 11.66％higher than control (P＞0.05).The activity of Acidity Phosphatase(ACP) and lysozyme in Cottonseed meal hydrolysates was significant higher than control(P＜0.05), the activity of Superoxide Dismutase(SOD), Acidity Phosphatase(ACP), Alkaline Phosphatase(ALP) and lysozyme was no-significance with the control.In finished experiments, based on analysis of chemistry and bio-active valve of plant protein hydrolysates, we have inclusion that cottonseed meal protein hydrolysates contain a lot of small molecular peptides and high bio-active valve, so we do next two experiments in which we used HPLC to analyses peptides of cottonseed meal protein hydrolysates and raising fish indoor, and then study how cottonseed meal protein hydrolystaes affect the group development, digestive performance and trypsin Mrna levels of Alloggnogeuetic crucian in order to evidence affecting mechanism.6 Analysis of protein and peptides components of cottonseed hydrolysates by HPLCIn this experiment, protein components in cottonseed and enzymatic hydrolysates of cottonseed were analyzed by general methods, gel filtration chromatography and high performance liquid chromatography (HPLC). It showed that: soluble proteins were increased by 1 time and free amino acids were increased by 2.9 times after been hydrolyzed by subtilisin; two main peaks were found by chromaography column of sephadexG-50, proteins in the cottonseed's peak were more than those in hydrolysate's. The molecular weight of the cottonseed and hydrolysate's two peaks were higher than 7000Da with a standard curve, but protein concentition of hydrolysates was less than that of cottonseed meal:the HPLC method was used to determine that the hydrolysates contained more proteins than cottonseed, the contents were decreased 7.71％, but small peptides(2-10 amino acids) were increased 24.73％, and tree amino acids were also increased. Animals can absorb small peptides and free amino acids firsthand ,so cottonseed hydrolysates were better for animals contrast with cottonseed。7 Effect of cottonseed meal hydrolysates on group, trypsin's activity and expressing level of trypsinogen's mRNA of Allogygenetic Crucian CarpTo explore the effect of Cottonseed meal hydrolysates and additive quantity in diets on Allogynogenetic Crucian's growth, trypsin's activity and expressing level of trypsinogen's mRNA, and to study the relationship of proteinase's activity and Allogynogenetic Crucian's productive performance and biochemistry index. Firstly, we use the Asl.398 Neutrase to hydrolyze the Cottonseed meal hydrolysates, secondly, use the hydrolysates to feed Allogynogenetic Crucian (weight30±2g) raised in the inner flowing cultivating system by 1.5 and 3.0 percentage for 65 days, finally, measure the fish's growth index, ADC of nutriment, the proteinase's activity and the expressing level of trypsinogen's mRNA. The result stated that, the body weight of 1.5％and 3.0％groups fish were higher then the control by 8.96％and 44.24％respectively raising for 35 days, and significantly different (p＜0.05), furthermore after 65 days the higher was 33.19％and 69.62％(p＜0.01), and the expressing level of trypsinogen's mRNA were also enhanced, and all the data suggest the 3.0％was the best level, and the correlation of the Allogynogenetic Crucian's growth and expressing level of trypsinogen's mRNA ,simultaneously the hydrolysates could accelerate the trypsin's activity and ADC of nutriment.
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