Production Of Glucose Oxidase By Fermentation

(3-D-glucose can transformed into gluconic acid by GOD with oxygen molecule as electron attachment carrier with a by-product of hydrogen peroxide?H₂O₂?at the same time.Due to its wide applications in foodstuff,pharmaceutical industry,chemical field,biotechnology,drinks,medical chemistry and other related field,microbial glucose oxidase is receiving more and more attention.New utilization of glucose oxidase?GOD?in biosensing equipment have promoted the market demand in the last years.Utilization of glucose oxidase in many domain and as a detection enzyme are having an promoting impact on biological processing.In this study,a bacterial strain of Pichea pestoris was used to study the strain.The fermentation conditions of the strain were studied in shake flask and fermentor.The method of measuring the concentration of methanol in liquid and the method for measuring the activity of glucose oxidase were also studied.Refer to relevant literature,combined with laboratory conditions,studied the method of determining the concentration of methanol in solution and determination of glucose oxidase activity.Shake flask condition:Using YPD as seed medium,the best seed age was 30h.The optimum medium for glucose oxidase fermentation was optimized by orthogonal test:Beef extract 1.0%?Peptone 2.0%?YNB 1.34%?Methanol concentration at 2%?Calcium carbonate concentration is 1%.Fermentation procass comdition:Optimum fermentation tempareture:30??beginning pH value:6.5?Shaking speed:210 r·min^-1 The enzyme production level reached 31.78U · mL^-1.5L fermentation tank:The initial biomass is 90g · L^-1,Methanol induction at a constant flow rate of 5.8 ·?L · h?^-1,Culture 128 h,The tallest enzyme activety can reach at 605 U ·mL^-1.