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Method for separating and purifying phytase with simpleness and high efficiency

A technology for separation and purification of phytase, applied in the direction of hydrolytic enzymes, etc., can solve the problems of low adsorption capacity, difficulty in use, and high price of affinity chromatography columns

Inactive Publication Date: 2009-09-16
广州市开恒生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the steric hindrance of the solid-phase affinity chromatography adsorbent, the adsorption capacity is low, and the preparation of the affinity chromatography column is expensive and costly, so it is difficult to use in industrialized large-scale production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Take 5 kg of phytic acid, add deionized water to 25 kg, mix well, add all to 1000 liters of centrifuged supernatant to make the concentration 0.5%, 25 ° C, pH 5.0, stir and mix. 3%MgSO 4 10 kg was added to the above mixture to make its concentration 0.03%, and stirred and mixed. The mixture is centrifuged by a centrifuge to obtain phytase wet enzyme powder. The wet enzyme powder is dried by a vacuum rake dryer to obtain a phytase powder preparation. The purity of phytase is 81.3%, and the yield is 76.6%.

Embodiment 2

[0014] Take 3.2 kg of sodium phytate, add deionized water to 16 kg, mix well, add all to 800 liters of centrifuged supernatant to make the concentration 0.4%, 30 ° C, pH 6.0, stir and mix. 5% CaCl 2 8 kg was added to the above mixture at a concentration of 0.05%, and stirred to mix. The mixture is centrifuged by a centrifuge to obtain phytase wet enzyme powder. The wet enzyme powder is dried by a vacuum rake dryer to obtain a phytase powder preparation. The phytase has a purity of 79.5% and a yield of 78.4%.

Embodiment 3

[0016] Take 6 kg of calcium phytate, add deionized water to 30 kg, mix well, add all to 1000 liters of centrifuged supernatant, 28 ° C, pH 7.2, stir and mix. Add 10 kg of 6% NaCl to the above mixture with a concentration of 0.06%, and stir to mix. The mixture is centrifuged by a centrifuge to obtain phytase wet enzyme powder. The wet enzyme powder is dried by a vacuum rake dryer to obtain a phytase powder preparation. The phytase has a purity of 78.0% and a yield of 80.5%.

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Abstract

The invention provides a method for separating and purifying phytase with simpleness and high efficiency. The method comprises the following steps of: centrifugating fermentation liquor producing the phytase by a centrifuge, collecting supernatant, utilizing substrate applied by the phytase, namely phytic acid (or phytate), as affinity chromatography adsorbent, directly adding the phytic acid (or phytate) into the supernatant, stirring fully and evenly, and absorbing phytic acid (or phytate) and the phytase with specificity. Due to strong sequestering power, the phytic acid (or phytate) generally forms insoluble salts with minerals of calcium, magnesium, sodium, zinc, iron, copper, manganese and the like. At the moment, the adding of inorganic salt containing the metal ions can precipitate the combined body of phytic acid and phytase, and the precipitates can be collected by the centrifuge and is dried by a drier so as to obtain the preparation of phytase powder. The method has the advantages of simple and convenient separation and purification, high yield, high purity, low cost and the like.

Description

technical field [0001] The invention relates to a simple and efficient method for separating and purifying phytase. The invention particularly relates to a kind of direct separation and purification of phytase from microbial submerged fermentation liquid. The method has the advantages of simple separation and purification, high yield, high purity and low cost. Background technique [0002] Common methods for separating and purifying enzymes from microbial submerged fermentation broths are: [0003] (1) Precipitation separation: By changing certain conditions, the solubility of some solutes in the solution is reduced, so that they can be precipitated from the solution to achieve the purpose of separation from other solutes. There are many methods of precipitation separation, such as salting out precipitation, isoelectric point precipitation, organic solvent precipitation, composite precipitation, selective denaturation precipitation, etc. The yield of the purified enzyme b...

Claims

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Application Information

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IPC IPC(8): C12N9/16
Inventor 邓开健何倩松
Owner 广州市开恒生物科技有限公司
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