Preparation Of Phospholipase C And Study On The Removal Of Rapeseed Detached

By growing the Bacillus cereus strain Shenzhen754-1in growth medium of richnutrition.we obtain preparation of phospholipase C（PLC） using microbial fermentationand optimize the fermentation conditions. We detected the activity of phospholipase Cproduction by egg yolk plate method and quantitatively measured the activity ofphospholipase C by the NPPC method. We also study some properties of phospholipase C.by analysising and discussing of the different reaction conditions， the effect of vegetableoil degumming by phospholipase C are detected.so we can provide experimentalparameters of oil degumming of phospholipase C in oil industry. In this paper， the mainresults are as follows：1. B.cereus Shenzhen754-1in shake flask fermentation，The best fermentation conditionare：the ratio of beef extract and peptone in medium10：1、culture temperature32℃、time24h、 speed180r/min.The content of the phospholipase C production by B.cereusShenzhen754-1prepareing reach29U/ml.2. obtaining the phospholipase C by Microbial method， the activity were detected byusing crude enzyme of the yolk agar method， the size of halo reflect the enzyme activitylevel，the bigger the halo is，the higher activity enzyme is. B.cereus754-1phospholipaseC was sensitive to heat， but also sensitive to pH. The optimum temperature is60℃in theNPPC reaction system， the optimum pH is7， the most stable activity is at about pH6.0~8.0.3. Analysising the different conditions of sulfate saturation and the settling time，comparing the effect of phospholipase C settlement by the size of halo in egg yolk plate asindex，the best conditions of the settlement is：ammonium sulfate saturation is40%， thesettling time of24h.3. Analysising the different conditions of the temperature、 time、pH、amount ofenzyme，we study on rapeseed oil degumming by the laboratory prepared phospholipaseC，and optimized degumming conditions the results are as follows： enzyme dosage 0.05%， temperature60℃， pH=7， time220min， the lowest phosphorus degummingoil residue is4.3mg/Kg.