Method for preparing oxalic acid decarboxylase by fungus

A technology of oxalate decarboxylase and fungi, applied in the direction of lyase

Inactive Publication Date: 2007-10-31
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Strict control of reaction cond

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  • Method for preparing oxalic acid decarboxylase by fungus
  • Method for preparing oxalic acid decarboxylase by fungus

Examples

Experimental program
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Embodiment 1

[0027] 1. Preparation of acid solution

[0028] The liquid acid can be made into a concentrated solution by pipetting the corresponding amount, and the solid acid can be made into a corresponding concentrated solution by weighing a certain amount of powder with deionized water. The concentration is generally about 1mol / L to 2mol / L. The oxalic acid solution was prepared with an ion concentration of 0.5M. The prepared acid solution was sterilized at 121°C for 20 minutes or sterilized by filtration before use.

[0029] 2. The amount and timing of acid addition

[0030] On the third or fourth day of bacterial growth, that is, when the growth is about to enter the logarithmic growth phase, the inducer is added. The amount of oxalic acid or oxalate added is such that the final oxalate ion concentration in the medium is 3-40 mmol / L. Other acids were added in such amounts that the pH of the culture medium dropped below 3.0.

[0031] 3. Trametes versicolor PRL 572 produces oxalate ...

Embodiment 2

[0047] 1. Preparation of acid solution

[0048] The liquid acid can be made into a concentrated solution by pipetting the corresponding amount, and the solid acid can be made into a corresponding concentrated solution by weighing a certain amount of powder with deionized water. The concentration is generally about 1mol / L to 2mol / L. The oxalic acid solution was prepared with an ion concentration of 0.5M. The prepared acid solution was sterilized at 121°C for 20 minutes or sterilized by filtration before use.

[0049] 2. The amount and timing of acid addition

[0050] On the third or fourth day of bacterial growth, that is, when the growth is about to enter the logarithmic growth phase, the inducer is added. The amount of oxalic acid added is such that the final concentration of oxalate ions in the medium is 5-40 mmol / L. Other acids were added in such amounts that the pH of the culture medium dropped below 3.0.

[0051] 3. Aspergillus niger (Aspergillus niger) ATCC 46890 pro...

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Abstract

The invention discloses a preparing method of oxalic acid decarboxylase with fungi, which comprises the following steps: (1) moist-heat-sterilizing organic acid or inorganic acid solution with high temperature at 121 deg. c; (2) setting the initial pH value of liquid medium of trametes versicolor (Trametes versicolor) or aspergillus niger (Aspergillus niger) and so on fungi at 5. 0-5. 5; setting the reacting temperature at 20-25 deg. c and air capacity at 20-30L/L . h; adding the acid solution at the third or fourth day with pH value at 0-3; centrifuging; collecting thallus cell; or filtering; getting thallus cell; eluting with o. 05-0. 2M and pH value at 3-3. 7 acetic acid buffer solution; collecting; freezing; grinding hypha; adding into acetic acid-sodium acetate buffer solution with 0. 05-0. 2M and pH value at 3-3. 7; grinding to pasty continually; centrifuging; getting oxalic acid decarboxylase liquid. This invention possesses low cost, which can be used to medical, food, beverage, agriculture and so on aspects.

Description

technical field [0001] The invention belongs to the technical field of biological enzyme preparation, in particular to a method for preparing oxalate decarboxylase by fungi. Background technique [0002] Oxalate decarboxylase (Oxalate decarboxylase, EC 4.1.1.2) is a manganese-containing enzyme that can catalyze the decomposition of oxalate to produce formic acid and carbon dioxide (see formula 1). The enzyme has a wide range of potential applications, such as removing oxalate precipitation in the pulp and paper industry, and can be used in clinical medicine to analyze the oxalate content of biological samples to diagnose hyperoxaluria and urinary stones / kidney stones, etc. In the food industry, it is used to analyze, control and reduce oxalic acid in food (such as spinach, chocolate, etc.) to produce low oxalic acid healthy food; in the beer industry, it can be used to control and degrade beer stone rich in oxalate; alumina smelting It can also be used in industry to deal w...

Claims

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Application Information

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IPC IPC(8): C12N9/88
Inventor 洪枫朱翠侠
Owner DONGHUA UNIV
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