Studies Of Chromatographic Technical In Trehalose Separation

The main subject of the conversion of trehalose synthase trehalose production, thefirst method for the fermentation engineering bacteria broken, using ultrasonic,lysozyme and high pressure homogenization bacterial release activity, the results showthat high-pressure homogenizer. The qualitative method broken bacterial short time, thecell is completely broken, releasing activity best results.After Bacterial cells broken, trehalose synthase is released. And maltose syrup as asubstrate for enzymatic transformation experiments. Experimental study on theoptimization of transformation conditions to determine. The trehalose synthase optimalpH of7.5; optimal reaction temperature of40°C; transformed completely the best timeis6hours; the substrate maltose concentration of30%, at this concentration, trehaloseconversion rate reached73.5%.The transformed sugar processing, mainly because of trehalose and maltose isextremely difficult to separate, part of the generation of glucose conversion, thedecision in which degradation of maltose to glucose, and then separating. Select atpH=4.5, temperature of60°C and200U/g enzyme plus the amount of glucoamylase for8hours after transforming the sugar maltose glucose degradation of all, no residue,trehalose retention value of99.90%.Before separation of sugar decolorization desalting treatment, the use of activatedcarbon for the saccharification liquid bleaching, optimum bleaching conditions: pH=5.0,temperature75°C, the dosage of2%(w/v) activated charcoal decolorization time60minutes. After desalting by decolorization, trehalose is almost no loss. Sugar, glucoseand trehalose, can be directly carried to the next stage of chromatographic separation.Filter out the separation effect the best A216Ca2+-type resin in the resin fromseveral possible by a large number of static shake flask screening and single columnelution test. Optimized single column separation conditions: elution of pure water, atemperature of60°C, injection volume10mL elution flow rate1mL/min reached themaximum separation R=0.56.By single column evaluation experiments mixed with sugar in the simulatedmoving bed system, the application of the triangle theory, found under linear conditionsand non-linear conditions completely separated. Obtained according to the completeseparation of the boundary conditions of the triangle, the theory SMB separation system operating parameters. The actual operation by simulated moving bed constantlyoptimize the theoretical parameters, optimal separation by the effect of: trehalose purityof97.6%, the average mass concentration of58.1g/L; glucose purity of98.8%, theaverage mass of71.2g/L After three cycles of loop stability, can be calculated trehaloseand glucose recoveries were83.2%and79.2%, respectively. The continuous directseparation of trehalose and glucose, the simulated moving bed can be fully applied tothe separation of trehalose, trehalose study was separated to provide a basis forindustrialization.Separation of trehalose crystallization experiments, the trehalose mother liquor wasconcentrated to60%when added3-fold volume of95%ethanol, stirred slowly tocrystallize under uniform size can be obtained, the purity of about98.4%trehalosecrystal recoveries94.2%.