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A method for separating and purifying acetoin in fermentation broth

A fermentation broth and acetoin technology, applied in the field of bioengineering, can solve problems such as excessive organic acids and unfavorable rectification, and achieve the effects of less water content, lower energy consumption, and less scaling

Active Publication Date: 2019-06-25
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The present invention aims at the problems that the upper phase obtained when acetoin in the fermentation broth is extracted by two aqueous phases formed by a hydrophilic organic solvent-salt contains more water, pigments, residual sugars, organic acids, and is unfavorable for subsequent rectification. A new method for extracting and separating acetoin in fermentation broth based on multi-stage countercurrent salting-out extraction technology with weakly polar organic solvents

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  • A method for separating and purifying acetoin in fermentation broth
  • A method for separating and purifying acetoin in fermentation broth
  • A method for separating and purifying acetoin in fermentation broth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 The influence of different organic solvents on the distribution of acetoin

[0034] Add 17.2 g of anhydrous sodium carbonate or 120 g of dipotassium hydrogen phosphate to 100 mL of the fermentation clear liquid containing 35 g / L of acetoin, and dissolve to obtain a salt solution after stirring. Take 4 mL each of the salt solution and the organic solvent (shown in Table 1) and place them in a graduated 20 mL test tube with a stopper. Vortex for 1 minute and let stand for 8 hours in a water bath at 20°C. The upper phase is rich in acetoin. The organic phase, the lower phase is the salt-rich aqueous phase. The distribution data of acetoin is shown in Table 1. It can be seen from the data in the table that for organic solvents with similar structures, such as alcohols and acetates, the recovery rate of acetoin decreases as the polarity of the solvent decreases. Using high-polarity organic solvents (propanol and butanol), the upper phase contains more water when the...

Embodiment 2

[0037] Example 2 Effect of ethanol content in solvent on acetoin distribution

[0038] Put 4mL of the fermentation clear liquid containing 60.5g / L acetoin, 2g of inorganic salt and 4mL of solvent in a 20mL test tube with stopper, vortex for 1min and let stand in a 24℃ water bath for 12h, separate layers, the upper phase is rich The organic phase of acetoin, the lower salt-rich water phase, the inorganic salt is dipotassium hydrogen phosphate or potassium pyrophosphate, the solvent is ethyl acetate or methyl tert-butyl ether, and the solvent contains 0~ 100% ethanol, test the comparison of the upper and lower phases (R), the partition coefficient (K) and recovery rate (Y) of acetoin in different extraction systems, such as figure 1 Shown. figure 1 The salting-out extraction systems of A to D are as follows: A, methyl tert-butyl ether-ethanol / potassium pyrophosphate; B, methyl tert-butyl ether-ethanol / dipotassium hydrogen phosphate; C, ethyl acetate-ethanol / Potassium pyrophosphate...

Embodiment 3

[0039] Example 3 The influence of salt dosage on the distribution of acetoin

[0040] Place 4 mL of the fermentation clear liquid containing 60.5 g / L acetoin, inorganic salt and 4 mL of organic solvent in a 20 mL test tube with a stopper, vortex for 1 min and let stand in a 24 ℃ water bath for 12 h. The organic phase of acetoin, the lower phase is the salt-rich aqueous phase, where the inorganic salt is dipotassium hydrogen phosphate or potassium pyrophosphate, and the organic solvent is ethyl acetate or methyl tert-butyl containing 20% ​​or 30% ethanol by volume Base ether, detect the comparison of the upper and lower phases (R), the partition coefficient (K) and recovery rate (Y) of acetoin in different salting-out extraction systems, such as figure 2 Shown. figure 2 The abscissa of A~D is the mass volume percentage of the added amount of inorganic salt (m, g) in the volume of fermentation broth (v, mL), figure 2 The salting-out extraction systems of A to D are as follows:...

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Abstract

The invention discloses a method for separating and purifying acetoin in fermentation liquor. The method includes the step of adding inorganic salt and a weak polar organic solvent are added into the fermentation liquor for extraction, wherein the addition amount of the inorganic salt accounts for 25-125% of the volume of the fermentation liquor, and the volume ratio of the organic solvent and the fermentation liquor is (1-2):1. The weak polar organic solvent is one of ethyl acetate, methyl tertiary-butyl ether, butyl acetate, dimethyl carbonate and diethyl carbonate. By means of the method, an extraction phase containing acetoin hardly contains pigment, residual sugar and organic acid impurities, an extraction solution can be directly rectified, and the scaling phenomenon is not likely to happen to a column reactor in the rectification process. Polarity of the solvent in use is small, the specific heat capacity is low, the upper phase water content is low, energy consumption for recycling is low, and acetoin loss in the recycling process is small. Operation is easy, and the method is an acetoin separation and extraction technology with industrial application prospects.

Description

Technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a separation technology of products in microbial fermentation broth, in particular to a method for separating acetoin in fermentation broth by using a multi-stage countercurrent salting-out extraction technology. Background technique [0002] Acetoin (3-hydroxy-2-butanone) is a natural fragrance and an important platform compound that exists widely in nature. It can be used to synthesize fragrance diacetyl and tetramethylpyrazine, and optically active α-hydroxy ketone derivative And a variety of pharmaceutical intermediates. [0003] Acetoin has two optical isomers, R- and S-. At present, commercial acetoin products are mostly synthesized by chemical methods, which are racemic mixtures of two optical isomers, and the products obtained by biological methods are mostly one of the isomers, so in recent years, fermentation methods Research on the production of acetoin has attracted m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07C45/80C07C45/78C07C49/17
CPCC07C45/78C07C45/80C07C49/17
Inventor 戴建英关文天马林会修志龙
Owner DALIAN UNIV OF TECH
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