Preparation Of Glycerylphosphorylcholine Enzymatic Catalyzed By Complex Phospholipase And Its Methyl Reaction Of Fatty Acid In Extract

This research studied enzyme hydrolysis of GPC in the water phase system with compositephospholipase. Glycerol phosphate choline and its stability by different separation andpurification method were aslo discussed in this research. The fatty acid in extract of compoundphosphatide enzyme preparation of glycerin phosphatidyl choline was used to esterificationreaction. We draw the following conclusions:(1) Enzyme hydrolysis with phospholipase A1and phospholipase A2can effectivelyreduce the reaction time and even improve the production rate of GPC. The optimal enzymatichydrolysis conditions determined by response surface optimization test was: substrateconcentration61g/L, enzyme hydrolysis temperature45.2℃, phospholipase A1additive amount16.86U/mL, phospholipase A2additive amount21.98U/mL, CaCl2adding amount5.18%,under this condition, yield of GPC has the optimal value of96.01%.(2) Enzyme hydrolysis with phospholipase A1and phospholipase A2can effectively reducethe reaction time and even improve the production rate of GPC. The optimal enzymatichydrolysis conditions determined by response surface optimization test was: substrateconcentration92.1mg/mL, enzyme hydrolysis temperature50.46℃, phospholipase A1additiveamount12U/mL, phospholipase A2additive amount12U/mL, CaCl2adding amount4.50mg/mL,under this condition, yield of GPC has the optimal value of95.34%.(3) Glycerol phosphate choline and its stability by different separation and purificationmethod were discussed in this research. GPC by enzyme hydrolysis was set as substrate, thenpurification effect of freezing crystallization, silica gel column chromatography and ionexchange resin method was compared. Silica gel column chromatography and ion exchange resinwere found to be an effective purification technology of GPC, the purity was98%. GPC in thepreparation of the aqueous system simplified the separation and purification steps of lecithin andLPC. GPC purified by silica gel column chromatography and ion exchange resin technology wasstrong; product quality standards meet the requirements of the quality of the same productsabroad.(4) The fatty acid in extract of compound phosphatide enzyme preparation of glycerinphosphatidyl choline was set as substrate in this research. Ultrasonic treatment was assisted to catalyze the esterification reaction, the optimal reaction condition was established by responsesurface method: methanol: the extract ratio of16.45:1, ultrasonic treated time of16.04min,ultrasonic power311.4W, dosage of NaOH1.90%, under this condition, the fatty acid methylester conversion has the optimal value of92.57%.