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Preparation, separation and impurity-removal purification methods of L-malic acid

A malic acid and seed technology, applied in the field of fermentation, can solve the problems of low L-malic acid fermentation yield, residual aflatoxin, increase enterprise cost, etc., and achieve the goal of increasing industrial added value, reducing mass transfer restrictions, and increasing yield. Effect

Active Publication Date: 2017-08-22
SHANDONG FUFENG FERMENTATION CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] With the gradual development of the amino acid industry, the fermentation production technology of L-malic acid has also made great progress, but the fermentation yield of L-malic acid is low and the conversion rate is not complete
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
In view of the above defects, the applicant's previous patented technology "a method for preparing L-malic acid" used Aspergillus oryzae fermentation to prepare malic acid, which adopted a mixed fermentation method and achieved a good yield of malic acid; the applicant's previous The patented technology "a new extraction process of L-malic acid" adopts the steps of neutralization, acidolysis, ion exchange and low-temperature concentration and crystallization to separate and purify malic acid. The purity of malic acid can reach 99%, but the steps are cumbersome, and high operating costs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A method for preparing, separating and removing impurities and purifying L-malic acid, comprising the steps of:

[0026] According to Aspergillus oryzae seed liquid (concentration is 1×10 10 CFU / mL): The fermenter culture medium is transferred to the fermenter at a volume ratio of 1:8 for cultivation at a temperature of 32°C for 40 h, then calcium carbonate and cetyltrimethylammonium bromide (CTAB) are added, and the carbonic acid The amount of calcium added is 80g / L, the amount of CTAB added is 40mg / L, and the fermentation is continued for 4 hours, then the temperature is controlled at 39°C, the pressure is 2 atmospheres, the heat preservation and pressure are kept for 6 hours, and the fermentation is stopped to obtain L-malic acid fermentation During the fermentation process, the pH is controlled at 6.2 by automatically feeding ammonia water; and the residual sugar is controlled at not less than 1.0% by feeding a glucose solution with a concentration of 200g / L; the cu...

Embodiment 2

[0029] Effects of various factors on the production of malic acid in Aspergillus oryzae fermentation broth:

[0030] Determination of L-malic acid: adopt 2,7-naphthalenediol colorimetric method, take 1.0 mL of sample solution, add 6.0 mL of analytically pure concentrated sulfuric acid, then add 0.1 mL 2,7-naphthalenediol solution, and then in 100 Heated in a water bath at ℃ for 20 min, took it out and cooled it to room temperature, and carried out colorimetric measurement at 385 nm, using distilled water as a reference to calibrate the zero point of the instrument. Use the standard sample to make a standard curve first, take the malic acid content as the abscissa, and the absorption value at 385 nm, that is, OD385, as the ordinate. Through the OD value of the unknown sample at 385 nm, the corresponding malic acid can be found on the standard curve. content.

[0031] Set up control groups, wherein, control group 1: do not increase the temperature and pressure, and the rest are...

Embodiment 3

[0035] 1. CTAB gradient test:

[0036]Taking Example 1 as an example, the impact of the addition time of CTAB on the amount of acid produced was detected, see Table 2 for details:

[0037] Table 2

[0038] time h 20 30 40 50 60 70 Malic acid output (g / L) 104.5 113.6 117.4 128.9 124.9 118.3

[0039] 2. Pressure gradient test: For the final 6 hours of fermentation, select 1-5 atmospheric pressure for testing, and the rest of the experimental process is the same as in Example 1. The specific fermentation results are shown in Table 3:

[0040] table 3

[0041] Atmospheric pressure 1 2 3 4 5 Malic acid output (g / L) 113.5 128.9 124.3 109.6 98.7

[0042] 3. Temperature gradient test: For the last 6 hours of fermentation, select 33-43°C for the test, and the rest of the experimental process is the same as in Example 1. The specific fermentation results are shown in Table 4:

[0043] Table 4

[0044] temperature...

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Abstract

The invention belongs to the technical field of fermentation and discloses a preparation, separation and impurity-removal purification method of L-malic acid. The methods comprise the following steps of: 1) fermentation; 2) transparent treatment; 3), separation and impurity removal; and 4) purification. The output of the malic acid prepared by the method is high, and the product yield and purity are high after impurity-removal purification.

Description

technical field [0001] The invention belongs to the technical field of fermentation, and in particular relates to a method for preparing, separating and purifying 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/46C07C51/42C07C59/245C12R1/69
CPCC07C51/42C12P7/46C07C59/245
Inventor 刘新利来凤堂贾召鹏刘建阳高翠娟薄文文范婷婷
Owner SHANDONG FUFENG FERMENTATION CO LTD
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