A process for producing malic acid by fermenting Aspergillus oryzae

A technology of malic acid and Aspergillus oryzae, which is applied in the direction of fermentation, microbial-based methods, microorganisms, etc., can solve the problems of low L-malic acid content, residual aflatoxin, and difficulty in industrial production, and achieve increased permeability, Effect of reducing mass transfer limitation and increasing yield

Active Publication Date: 2020-07-24
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.
[0004] 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

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] A kind of technology that adopts aspergillus oryzae fermentation to produce malic acid, it comprises the steps:

[0020] 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 200 r / min, and culturing for 12 hours to obtain a seed liquid, wherein the medium group of the seed tank Divided into: 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 6.0;

[0021] According to the volume ratio of seed liquid: fermenter medium is 1:10, transfer it to the fermenter for cultivation, at a temperature of 33°C, cultivate for 48h, then add calcium carbonate and cetyltrimethylammonium bromide (CTAB), calcium carbona...

Embodiment 2

[0025] A kind of technology that adopts aspergillus oryzae fermentation to produce malic acid, it comprises the steps:

[0026] 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 200 r / min, and culturing for 12 hours to obtain a seed liquid, wherein the medium group of the seed tank Divided into: 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 6.0;

[0027] According to the volume ratio of seed liquid: fermenter medium is 1:10, transfer it to the fermenter for cultivation, at a temperature of 33°C, cultivate for 48h, then add calcium carbonate and cetyltrimethylammonium bromide (CTAB), calcium carbona...

Embodiment 3

[0031] Effects of various factors on the yield of malic acid fermented by Aspergillus oryzae:

[0032] 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, you can check the corresponding malic acid content on the standard curve .

[0033] Set up a control group, wherein, control group 1: temperature and pressure are not increased during the fermentation process...

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PUM

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Abstract

The invention belongs to the technical field of microbial fermentation, and discloses a technology for producing a malic acid employing fermentation of aspergillus oryzae. The technology comprises the following steps of preparing an aspergillus oryzae seed solution, preparing the malic acid through fermentation, adding calcium carbonate and CTAB in the fermentation process, and adjusting the pressure intensity and the temperature. Through on permeability treatment on a bacterial strain, the technology is simple, the cost is low and the yield of the malic acid is improved.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a process for producing malic acid by fermenting Aspergillus oryzae. 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. Among them, L-malic acid widely exists in living organisms and participates in cell metabolism as a member of the tricarboxylic acid cycle; D-malic acid is not easily metabolized by the human body, and it is harmful to the human body if inhaled or swallowed. As a chiral source for chiral synthesis, Many important organic compounds can be synthesized. [0003] The production of L-malic acid is initially accompanied b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/46C12R1/69
CPCC12P7/46
Inventor 刘新利来凤堂李树标贾召鹏赵兰坤高翠娟范婷婷
Owner SHANDONG FUFENG FERMENTATION CO LTD
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