Method for immobilizing phospholipase by using cellulose acetate/polytetrafluoroethylene complex film

Method for immobilizing phospholipase by using cellulose acetate/polytetrafluoroethylene complex film

  • CN 102,304,504 A
  • Filed: 09/20/2011
  • Published: 01/04/2012
  • Est. Priority Date: 09/20/2011
  • Status: Active Application
First Claim
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1. method with cellulose acetate/polytetrafluoroethylcomposite composite film immobilized phospholipase is characterized in that realizing through following steps with the method for fixing of composite membrane Phospholipid hydrolase:

  • one, the preparation of composite membrane;

    obtain film-casting liquid with being dissolved in the 85g dimethyl formamide under 60 ℃

    of the 15g cellulose acetates.Poly tetrafluoroethylene is cleaned with the NaOH solution of 0.1mol/L and the HCl solution of 0.1mol/L earlier, use washed with de-ionized water then, after the vacuum-drying, be tiled on the clean sheet glass.Control thickness is coated in the poly tetrafluoroethylene surface with the cellulose acetate film-casting liquid with special scraper, after the reasonable time that at room temperature volatilizees is accomplished and done phase transformation, immerses deionized water and accomplishes the wetting phase conversion.Use washed with de-ionized water behind the 12h, seasoning is for use;

    Two, the immobilization of Phospholipid hydrolase;

    get some parts of 1cm 2Cellulose acetate/polytetrafluoroethylcomposite composite film in triangular flask, add Sodium phosphate dibasic/citrate buffer solution of a certain amount of enzyme liquid and pH=5.0 respectively, the whip attachment certain hour.The taking-up enzyme membrane was used deionized water rinsing after reaction finished, and was put in dry place and treated that its air-dry back mensuration enzyme is alive.Three, the mensuration of enzyme activity.Four, the optimization of fixing condition;

    respectively at enzyme liquid concentration 0.02g/mL~

    0.07g/mL, adsorption time 1h~

    6h, 45~

    70 ℃

    of temperature under the condition of stirring velocity 80r/min~

    280r/min, obtain best fixing condition through measuring the immobilized enzyme vigor.Five, adopt the response surface method to analyze the final optimal fixation condition of confirming is discussed.Six, further explain the advantage of utilizing cellulose acetate/polytetrafluoroethylcomposite composite film immobilized phospholipase through the SEM image of fixed enzyme membrane.

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