Preparation method of beta-alanine

A technology of alanine and aspartic acid, applied in the biological field, can solve the problems of poor enzyme stability and low enzyme activity

Pending Publication Date: 2020-09-29
GUANG AN MOJIA BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, existing methods have the problems of low enzyme activity and poor enzyme stability.

Method used

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  • Preparation method of beta-alanine
  • Preparation method of beta-alanine
  • Preparation method of beta-alanine

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[0019] The present invention provides a method for preparing beta-alanine, comprising: preparing a beta-alanine product from a reactant containing fumaric acid and ammonia water in the presence of a catalyst, wherein the catalyst comprises aspartic acid A catalytic composition of enzyme and L-aspartate-alpha-decarboxylase, and fumaric acid is added during the reaction, and the total amount of moles of fumaric acid added is equal to the initial molar amount of ammonia water in the reactant Subtract the initial molar amount of fumaric acid in the reactants.

[0020] The "initial molar amount" of fumaric acid or ammonia water refers to the initial molar amount of fumaric acid or ammonia water before the catalytic reaction begins. The "initial fumaric acid" weight in the reactants refers to the initial weight of fumaric acid added to the reactants prior to the start of the catalytic reaction. In certain embodiments, in the reactant, the initial molar ratio of the ammonia water to...

Embodiment 1

[0062] Fermentation culture of Escherichia coli engineering bacteria, inorganic salt fermentation medium formula: NaCl 1g / L; MgSO 4 -7H 2 O 1g / L; FeSO 4 -7H 2 O 0.02g / L; ZnSO 4 -7H 2 O 0.03g / L; CuSO 4 -5H 2 O 0.005g / L; (NH 4 ) 2 SO 4 4g / L; KH 2 PO 4 4g / L, sterilized at 121°C for 15 minutes, cooled to 37°C after sterilization, inoculated with 2% inoculum, fermented at 37°C, dissolved oxygen maintained at 30%-50%, ventilation volume 1VVM, Add ammonia water to adjust pH and provide nitrogen source, ammonia water to control pH at 7.5, feed glucose to provide carbon source, control residual sugar to 0.1-0.5g / L, and cultivate to OD 600 When = 30, the temperature was lowered to 30 °C and IPTG was added for induction, and the fermentation was terminated after 30 hours of induction and culture. 600 It reached about 110, and the wet weight of the cells after centrifugation reached about 120 g / L.

Embodiment 2

[0064] With reference to "Molecular Cloning Experiment Guide (3rd Edition)" (Science Press) and Li Y.et.al.Appl.Microbiol.Biotechnol.2017, 101, 6015-6021, the aspartic acid containing aspartic acid derived from Anoxybacillus flavithermus WK1 was prepared Escherichia coli engineering bacteria with acid enzyme (hereinafter referred to as "enzyme 1") and Escherichia coli engineering bacteria strains containing L-aspartate-α-decarboxylase (hereinafter referred to as "enzyme 2") derived from Bacillus thermotolerans, conventional LB The culture medium is fermented by the method of Example 1 and then centrifuged (rotating speed 5000 rpm, centrifugation time 5 min) to obtain wet Escherichia coli engineering bacteria for use.

[0065]Prepare 3L ammonium fumarate solution (100g fumaric acid / L), the initial molar amount of fumaric acid is 2.586mol, the initial molar amount of ammonia water is 5.172mol, add 3g E. coli engineering containing enzyme 1 derived from Anoxybacillus flavithermus ...

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Abstract

The invention provides a preparation method of beta-alanine. The method comprises the following steps of: preparing a beta-alanine product from a reactant containing fumaric acid and ammonia water inthe presence of a catalyst, wherein the catalyst comprises a catalytic composition comprising aspartase and L-aspartate-alpha-decarboxylase, and fumaric acid is added during the reaction, and the total molar amount of the added fumaric acid is equal to the amount obtained by subtracting the initial molar amount of fumaric acid in the reactant from the initial molar amount of ammonia water in the reactant.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing beta-alanine by enzyme catalysis. Background technique [0002] β-Alanine, also known as β-aminopropionic acid or 3-aminopropionic acid, is a β-type non-protein amino acid that exists in nature. β-Alanine is a multi-purpose organic synthetic raw material, mainly used for the synthesis of pantothenic acid and calcium pantothenate, carnosine, sodium pamidronate, balsalazide, etc. It is widely used in medicine, feed, food and other fields. large market demand. [0003] At present, the production methods of β-alanine are divided into two categories: chemical synthesis method and biological method. Because chemical synthesis of β-alanine has been studied earlier and the technology is mature, it is still the main production method at home and abroad. [0004] There are two main ways to produce β-alanine by chemical method. One is the reaction of acrylon...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/06
CPCC12P13/06
Inventor 苏金环曾聪明邱贵森刘文杰
Owner GUANG AN MOJIA BIOTECHNOLOGY CO LTD
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