Separation method for L-malic acid

A separation method, malic acid technology, applied in the field of microbial fermentation, can solve the problems of difficult industrial production, low sugar utilization rate, complex process route, etc., to reduce the production of aflatoxin, avoid slow proliferation of strains, improve The effect of fermentation efficiency

Active Publication Date: 2018-07-20
SHANDONG FUFENG FERMENTATION CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Later, L-malic acid was produced by the fermentation of carbohydrates. However, the traditional fermentation method for producing L-malic acid takes a long time to cultivate, consumes a lot of energy, has low acid production rate, and the utilization rate of sugar is not high, so it is difficult to realize industrial production.
With the gradual development of the amino acid industry, the fermentation production technology of L-malic acid has also made great progress, but the industrial average conversion rate of L-malic acid is 70%-80%, and there are still 20%-30% fumaric acid residues. The content of L-malic acid in the reaction solution is relatively low, about 100g / L. At present, there is no large-scale production method of L-malic acid in China.
The applicant's previous invention patents "a method for producing L-malic acid by direct fermentation" and "a new process for extracting L-malic acid" have greatly increased the production of malic acid and sugar Acid conversion rate, but Aspergillus flavus will produce a certain amount of aflatoxin, and aflatoxin will remain in the product. The subsequent separation steps are cumbersome, which increases the cost of the enterprise
The purity of malic acid in the fermentation broth is relatively low, and the process route for separating malic acid from the fermentation broth is complicated, including multi-step acid hydrolysis, filtration, refining, crystallization and other steps, resulting in low product yield and low purity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A method for separating L-malic acid, comprising the steps of:

[0025] 1) Aspergillus oryzae (Aspergillus oryzae) bacteria solution (concentration 1×10 8 CFU / mL) according to the inoculum amount of 8% (volume ratio) is inserted in the seed jar and cultivated, and at temperature, be 33 ℃, shaker speed is 200r / min, cultivate 16 hours and obtain Aspergillus oryzae seed liquid, wherein, the seed jar The medium components are: 3g sucrose, 0.2g sodium nitrate, 0.5g ammonium sulfate, 0.1g potassium dihydrogen phosphate, 0.1g dipotassium hydrogen phosphate, 0.1g manganese sulfate monohydrate, 0.01g ferrous sulfate heptahydrate, magnesium sulfate 0.01g, sodium chloride 0.01g, pH 6.0;

[0026] 2) Aspergillus parasiticus bacteria solution (concentration of 1×10 8 CFU / mL) according to the inoculum amount of 6% (volume ratio) is inserted in the seed tank and cultivated, at a temperature of 32 ° C, the shaking table speed is 200r / min, cultivated for 24 hours to obtain Aspergillus ...

Embodiment 2

[0030] A method for preparing L-malic acid, comprising the steps of:

[0031] Aspergillus oryzae (Aspergillus oryzae) bacterial liquid (concentration is 1 × 10 8 CFU / mL) according to the inoculum amount of 8% (volume ratio) into the seed tank for cultivation, at a temperature of 33 ° C, a shaker speed of 200r / min, cultivated for 16 hours to obtain a seed liquid, wherein, the medium of the seed tank The components are: 3g sucrose, 0.2g sodium nitrate, 0.5g ammonium sulfate, 0.1g potassium dihydrogen phosphate, 0.1g dipotassium hydrogen phosphate, 0.1g manganese sulfate monohydrate, 0.01g ferrous sulfate heptahydrate, 0.01g magnesium sulfate , sodium chloride 0.01g, pH value 6.0;

[0032] According to the volume ratio of seed liquid: fermenter medium of 1:8, transfer it to the fermenter for cultivation, at a temperature of 33°C, cultivate for 3 days, then add calcium carbonate, maintain the concentration of calcium carbonate at 80g / L, continue to ferment for 3 days, and stop th...

Embodiment 3

[0037] The present invention detects the influence of calcium carbonate on the production of L-malic acid. Calcium carbonate was not added on the third day of fermentation, and the others were the same as in Example 1. After testing, the output of L-malic acid was 143.5 g / L, and the residual sugar content was 1.9 g / L.

[0038] The present invention also detects that the content of aflatoxin in the fermented liquid obtained by using the applicant's previous patented technology "a method for producing L-malic acid by direct fermentation" is 12.69ug / L; The aflatoxin content in the liquid is less than 0.5ug / L.

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Abstract

The invention belongs to the technical field of microbial fermentation, and discloses a separation method for L-malic acid. The method comprises the following steps: 1) preparation of an Aspergillus oryzae seed liquid; 2) preparation of an Aspergillus parasiticus seed liquid; 3) fermentation; and 4) centrifugation, filtration and ion exchange. The L-malic acid product separated from a fermentationbroth has a high yield and a high purity, and the method has the advantages of simple operation process, low production cost and realization of large-scale production.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a separation method for L-malic acid. Background technique [0002] Malic acid, also known as hydroxysuccinic acid, hydroxysuccinic acid or 1-hydroxyethanedicarboxylic acid, has the molecular formula C 4 h 6 o 5 , molecular weight 134.09, structural formula HOOCCHOHCH 2 COOH. Malic acid has two optical isomers, dextrorotatory (D-type) and levorotatory (L-type), and three types of DL-type racemic products. L-malic acid is widely used. [0003] L-malic acid is an important component of natural fruit juice. It has a soft taste and a special fragrance. It does not damage the mouth and teeth. It is beneficial to the absorption of amino acids in metabolism and does not accumulate fat. It is a new generation of food sour agent and is well-known in the biological and nutritional circles. As "the most ideal food sour agent". It is currently replacing citr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/46C07C51/42C07C59/245C12R1/69C12R1/66
CPCC07C51/42C12P7/46C07C59/245
Inventor 赵兰坤刘新利刘元涛贾召鹏王俊铭王婷范婷婷袁军薄文文
Owner SHANDONG FUFENG FERMENTATION CO LTD
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