The Study On Lactate Flux Control Analysis And Cytokine Network Of Spermophilus Dauriucus In Different Physiological Condition

Metabolic Control Analysis(MCA)is a quantitative analysis theory and an experimental methodology with the systemic view for metabolic regulation and control.Metabolic Control Theory(MCT)suggests that metabolic flux is often controlled by several steps of pathway,and control step occurs unnecessarily on irreversible step,so-called “rate-limiting step”,it's possible that reversible step also plays an important role in the control of flux.However,the theory has certain limitations,for instance,it's quite difficult to calculate the flux control coefficient,and operating on the enzymes of pathways at gene level is needed to obtain a disturbing effect on enzyme activity,which is time-consuming and laborious.Metabolic control analysis theory(MCT)had lead a landmark development of metabolic engineering as soon as its establishment and,immediately,was widely applied in fundamental research and production,but there still exist two challenges which greatly restrict the development and application of MCT.The two“tough guys” are the acquirement of the disturbance means of enzyme activity and the calculation of flux control coefficients(FCCs)were two key problems of metabolic control analysis,which often need gene manipulation and complicated calculation.Temperature is an important environment factor to the metabolism of most organisms.In this topic,temperature was chosen as a disturbing means to enzyme activity for the first time,and the lactic fermentation pathway was researched,employing a relatively simple method.The results indicated that PDHC,ICDH,a-KGDHC and MDH had relatively higher control effects on lactate flux in this pathway,both of them reached 70%,and the effect of PDHC and MDH was 0.28599,0.209252.Oppositely,phosphofructokinase(PFK)had only a small value,even appeared a slightly negative control,which was accordant with the standpoint of MCT.Simultaneously,method of MCT was also used,and the correlation coefficient of the two results was 0.851,correlation was significant at 0.01 level(2-tailed),which illuminated that it was effective and feasible to determine the control and regulation of enzyme on the metabolic pathway flux,employing the method in this research.Some difficult problems suffered by MCT had been avoided in this new method,which hopefully becomes a simpler way for researching the control of metabolic flux.According to the Model showing regulation of the switch from carbohydrate to fatty acidsduring hibernation,we offer and advised our Model of accommodation mode1 of lactate.