| In order to study the expression of enzymes related to carbohydrate metabolismof fish. A preliminary study on hepatopancreas glucose metabolism-related enzymeexpression jade perch (Scortum barcoo) was conducted using two-dimensionalelectrophoresis and mass spectrometry from the proteomic level. Using corn starch ascarbohydrate source, nine experimental diets were formulated, which were dividedinto group A (fixed protein source), group B (fixed the sum of the carbohydrate sourceand protein source), and group C (fixed carbohydrate source). Each group was dividedinto three different groups based on the content of carbohydrate or protein. Each dietwas randomly assigned to triplicate groups.Breeding experiment results: group of A2has the weight of gain, relative weightof gain rate, protein efficiency ratio and specific growth rate, and group of A3is thelowest; the group of B3has the highest protein efficiency ratio, the group of C3is thelowest, Group of A2has the lowest Feed conversion rates and group of C3has thehighest. Each individual indicator has significant differences between the highest andlowest values (P <0.05), we can see the group of A2feed has the best utilization effect,and it has the best growth status, its dietary protein content in the feed is fixed at34.47%, digestible sugar content is30.26%. group of C3feed is the worst. On theweight of gain,A2group has no significant differences with group of C3, but hassignificant differences with B1group, and has significant difference with the othergroups,A2has no significant difference with B2in relative increase of weight gainrate and specific growth rate, but it has significant difference with B1and A1, and hasa very significant difference with the other groups.Electrophoresis results of experiments:The best schizolysis results were observedin lysate formula I (8M Urea,2M Thiourea,0.5%(w: v) of CHAPS,40mMTris--Base,1%DTT,2%(w: v) Pharmalyte pH3-10,1mM PMSF) with the ratio oflysate and sample liquid1:3, as well as.bubble swelling liquid was swelling buffer I(8M Urea,2M Thiourea,0.5%of CHAPS,0.5%ampholyte,0.02%bromophenol blue,1%of DTT). The best isoelectric focusing effect was found in isoelectric focusingprogram I (S1:0-500(V),500V· h. S2:500V,500V· h, S3:500-3500V,10000V· h,S4:3500,50000V· h. S5:3500-5000,8000). The results of mass spectrometry experiments: six kinds of enzymes related toglucose metabolism are detected by mass spectrometry: fatty acid binding protein,Adenine phosphoribosyltransferase, acetylglutamate kinase, ATP-citrateb synthase,catalase, Glyceraldehyde-3-phosphate dehydrogenaseA.They are different in theexpression of significantly.Fatty acid binding protein in group A, a1-a3, declined with the feed carbohydrate(starch) increased gradually,and it significantly increased in group B with the feedcarbohydrate (starch) increased,and it has irregular changes in Group C along with theincrease in protein (fish meal). C1, when the feed sugar (starch) and protein (fish meal)content is relatively low, binding protein expression level is the highest, followed byb3carbohydrate (starch) content of the highest and the lowest content of protein (fishmeal); when the carbohydrate (the highest starch) content of the lowest content ofprotein (fish meal), the expression of the fatty acid binding protein was the lowest.Adenine phosphoribosyl transferase enzyme: in group A which has the sameprotein content, phosphoric acid adenine ribosyltransferase enzyme expression levelsdeclined with the carbohydrate (starch) content increased (a1-a3), in group B,high-protein sugar group b1adenine ribosyltransferase enzyme expression is lowerthan the high-sugar low-protein group b3, aiso lower than the middle group b2, b1<b3<b2,in group C, low protein group adenine phosphate ribosyltransferaseexpression is lower, the middle group and the high-protein group is almost flat. b1,named the lowest carbohydrate group with the highest content of the protein, theadenine ribose transferase expression was the lowest; and low-carbohydrate group a1was the highest.Acetylglutamate kinase: in group A, acetylglutamate kinase showed a cleardownward trend with the rise of the feed carbohydrate (starch) content; in group Bacetyl glutamate kinase showed slightly a downward trend along with thecarbohydrate (starch) content increased, in group C, acetylglutamate kinase showed aclear upward trend with the rise of the feed carbohydrate (starch) content.ATP-citrate synthase: group A showed a clear upward trend while group Bshowed a slightly upward trend with carbohydrate (starch) content increased, but theoverall expression of group B was significantly less than in group A, in group C with the increase of protein content in the feed, the low protein group a1has the highestexpression, significantly higher than the middle group and the high-protein group,middle group c2ATP citrate synthase expression was slightly higher than with thehigh protein group c3.Catalase: in group A, catalase showed a downward trend with the rise of the feedcarbohydrate (starch) content (a1-a3),in group B: b1with the low carbohydrate highprotein had the highest expression of catalase, was significantly higher than b2intermediate group and b3with high-carbohydrate and low protein, b3with the highcarbohydrate and low protein group was slightly higher than intermediate group b2. Ingroup C: catalase expression of the overall showed a upward trend,c1<c2<c3, but c2was significantly higher than c1and c3slightly higher than c2.Glyceraldehyde3-phosphate dehydrogenase A: in group A, glyceraldehyde3-phosphate dehydrogenase A showed a clear downward trend along with the the rise offeed carbohydrate (starch) content,in group B, glyceraldehyde3-phosphatedehydrogenase A expression in b3was higher than the intermediate group b2andhigh-protein low-sugar group b1with the the rise of feed carbohydrate (starch)content, b3> b1> b2.In group C,glyceraldehyde3-phosphate dehydrogenase Aexpression showed a clear upward trend with the the rise of feed carbohydrate (starch)content. |
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