Research On The Experiment Of C₈APG' Synthesie Catalyzed By Immobile Glucosidase And C₈APG' Purification

Oetyl Poly gucoside(C8APG) is one kind of new-style nonionic surfactant (APG), not only has the characteristics of ordinary nonionic surfactant but has the characteristics of anionicsurfactant, It was mainly used in the filed of food, medicine, biology, cosmetics and detergent, etc. At present, the chemical method of synthesis of C8APG has been widely studied, but the purity of product is lower, process complex, high production cost, safety unwarranted, so it cannot be applicated in the flied of food, medicine, etc. The synthesis of APG through immobilized glucosidase which has the characteristics of mild reaction condition and simple process and the products have high purity, they can be biodegraded drastically, the immobilized glucosidase can be reused, low production cost. Therefore, the synthesis of APG by immobilized glucosidase will absorb more and more attention. However, no matter which way will be used to synthesis APG,the high carbon alchol will be excessive.The extra high carbon alchol eight affect the performance and quality of produce or limit its field of application.so,it is very important to eliminate high carbon alchol.The purpose of this paper is the reseach of technics of the fixed enzymatic synthesis of C8APG which be studied lessly at present but has higher value in research field, and the technics of separation and purification. The conclusions as follows:1.We studied the process parameters of synthesis of C8APG by fixed enzymatic through single factor and spherically symmetric design , and the conclusion:(1)By the method of ACA microcapsule to fixβ-glycosidase enzymes, And through colorimetric method (For nitro phenyl -β-D-rkatsiteli grapes as the substrate) to determine the enzymes living of free enzymes and fixation respectively. Free enzymes 24.27 u/g, unusedly fixed enzymes22.40 u/g, usedly fixed enzymes 21.45 u/g.(2)Synthetic reaction, with the rising of temperature,the efficiency of conversion of glucose of reaction system first increases, then decreases. in 55℃, it reach maximum, there has been some fixed enzymes Lost activity when the temperature reaches 60℃.(3)The ratio of atalyst and glucose has a influence on the efficiency of conversion of glucose of reaction system, the time of reacting system reach reaction equilibrium. The efficiency of conversion of glucose as the ratio increasing and increase gradually, the efficiency of conversion of glucose changed subtly when the ratio bigger than 1:5. (4)The ratio of glucose and octyl alcohol has more influence on the efficiency of conversion of glucose of reaction system,little influence on the time of reacting system reach reaction equilibrium.The efficiency of conversion of glucose as the ratio increasing and increase gradually, the efficiency of conversion of glucose changed subtly when the ratio bigger than 1:6.(5)The efficiency of conversion of glucose as the increase of the water percentage first greaten after decrescent,it reach the maximal when the water percentage is 10%.(6)The pH of water has little influence on the efficiency of conversion of glucose of reaction system,the time of reacting system reach reaction equilibrium.(7)With X₁(Time), X₂(Temperature), X₃( The ratio of octyl alcohol and glucose), X₄( The ratio of atalyst and glucose)the four main influence factors as the research object, by spherically symmetric design,draw a regression mathematics models: Y=24.978+0.133X₁+1.875X₂+2.145X₃+2.205X₄+0.005X₁X₂-0.004X₁X₃-0.002X₁X₄+0.034X₂X₃+0.030X₂X₄-0.055X₃X₄-0.002X₁²-0.025X₂²-0.215X₃²-0.241X₄²To analyse the regression model ,we can get the conclustion: there are positive interaction between each factors. The optimal conditions is the reaction time 72.0h, the reaction temperature 54.0℃, The ratio of octyl alcohol and glucose7.7:1, The ratio of atalyst and glucose6.8:1.2.Optimize the process parameters that using column chromatography to separate and purificate C8APG by single factor and orthogonal experiment, draw conclusions as follow:(1)The separated effect will be better as the column`s diameter more bigger, the purity can be when the diameter isφ30mm,the mass fraction of n-octyl alcohol in the production is 176447ppm.(2)The solubility of octyl alcohol in the organic solvent ,the higher the bette; to the C8APG , the lower the batter. The purity of production is better when the eluent is Mineral ether/Acetic ether(1:1), the mass fraction of n-octyl alcohol in the production is 249579ppm.(3)The adsorption effect to n-octyl alcohol by column material is more little, but more strong to C₈APG, the effection of separated will be better. When the column material is alkaline alumina, the mass fraction of n-octyl alcohol in the production is 202124ppm.(4)By poor analysis shows that, the primary and secondary order of the influence on the test index by each factor is C,A,B, the optimal combination of each factor is A₁B₁C₂. Namely, the column material is activated carbon, solvent elution is B (Mineral ether/Ethyl acetate = 1:1), column specifications is diameterφ20×300.(5)From the analysis of variance we can come to a conclusion, factor A ,Cand B are significant factors, the significance level of AandB is 0.05, the significance level of C is 0.01. From the point estimation and interval estimation of optimum assembly wo can see that, the confidence interval is [0.2529-0.4805], the degree of confidence is 95%.3 . Take oxidation decoloring to the C8APG, and structural analysis,and performance testing.We can draw conclusions as follow:(1)To choice the influence factors of hydrogen peroxide bleaching by single factor experiment, when the solution concentration of C8-APG is 0.3%, the dosage of hydrogen peroxide is 40%, temperature is 80℃and decoloring time is 90min,we can make the extinction coefficient of product less than 0.01.(2)By TLC to analysize the properties of C8APG, the result indicates that there are not remaining octly acohol and glucose in the product.(3)By GC to analysize the properties of C8APG, t he result indicates that the concentration of octly acohol is 0.01%.(4)By IR to analysize the properties of C8APG, the map is according with the infrared spectra of C8-APG, further verified test the results.(5)The spunescence of C8APG is better, rich foam, exquisite and stablity; when the concentration is 40%, the height of foam can keep in 20mL standing about 20 minutes.(6)The stability of C8APG in the hard water is worse, they foam stability decreasing as the calcium ion concentration increasing, when the calcium ion concentration is 9mmol or 12 mmol, the height of foam reach only about 10mL.(7)The hydrotropy of C8APG to the vegetable oil will be batter as the solution concentration of C8APG increasing, when the concentration of aqueous solution of production is more than 0.3%, the area of oil measure is almost disappear.(8)Emulsification is general, emulsification will be better with the concentration of aqueous solution of production increasing.(9)Surface tension is lower, the surface tension can reach 0.0278 N/m when the concentration of C8APG is 0.1%.(10)The concentration of critical micelle is 0.026%.(11)The relative viscosity of C8APG is low. Innovation points:(1)By single factor and spherically symmetric design to optimize the process of ACA microcapsule immobilized enzyme catalytic synthesis C8APG, the technological parameters are obtained.(2)By single factor and orthogonal test to optimize the process of column chromatography separate and purificate C8APG, the technological parameters are obtained.