Method for producing L-sodium malate through direct enzymatic conversion

A technology of sodium malate and calcium malate, applied in fermentation and other directions, can solve the problems of restricting the wide application of L-sodium malate, consumption of large sulfuric acid, low concentration, etc., achieving significant industrial application value and environmental benefits, and reducing the degree of reverse reaction. , the effect of reducing production costs

Inactive Publication Date: 2018-10-16
THE GREAT WALL BIO CHEM ENG CO LTD
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Problems solved by technology

[0003] However, compared with the production cost of DL-sodium malate, the current commercial production technology of L-sodium malate is to prepare L-sodium malate by neutralizing L-malic acid and sodium carbonate. There are following disadvantages: (1) generate a large amount of calcium sulfate as by-product in this process, consume a large amount of sulfuric acid during acidolysis, and the consumption of calcium carbonate is also higher; (2) adopt L-malic acid to carry out And reaction when preparing L-sodium malate, because the price of L-malic acid as raw material is higher, therefore cause the production cost of L-sodium malate higher, thereby restricted the extensive application of L-sodium malate; ( 3) When the above process covers the production steps of L-malic acid, since this step involves concentration, crysta...

Method used

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  • Method for producing L-sodium malate through direct enzymatic conversion
  • Method for producing L-sodium malate through direct enzymatic conversion
  • Method for producing L-sodium malate through direct enzymatic conversion

Examples

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Effect test

Embodiment 1

[0077] The preparation of embodiment 1 L-sodium malate

[0078] Preparation of Calcium Fumarate Suspension

[0079] Preheat 2.5L of pure water to 55°C, add 348g of fumaric acid, then slowly add 300g of calcium carbonate, stir and mix with a mechanical stirrer, keep warm to neutralize the two, and monitor the concentration of fumaric acid at any time during the reaction. Content, when its content is lower than 10g / L, reaction finishes, obtains 3L calcium fumarate suspension.

[0080] Enzymatic Transformation Production of Calcium L-Malate

[0081] According to the amount of 300U enzyme activity / 100mL substrate, add the recombinant Escherichia coli introduced and expressing fumc gene into the calcium fumarate suspension prepared in the previous step as fumarate hydratase (calculated by enzyme activity) to control the enzymatic reaction The temperature is 37° C., and the enzymatic reaction is carried out under stirring at a stirring speed of 200 rpm, and 3.5 L of calcium L-mala...

Embodiment 2

[0085] The preparation of embodiment 2 L-sodium malate

[0086] In this example, except adopting 348g fumaric acid and 138g calcium carbonate and adding calcium hydroxide to adjust the pH to 6.8, the method described in Example 1 was used to prepare L-sodium malate, wherein, as a solid precipitate The calcium carbonate of the product is recovered and recycled in the preparation of calcium fumarate suspension.

[0087] The average productive rate of the L-sodium malate of this embodiment is 28.34g / (L h), and the catalytic efficiency of fumaric acid hydratase is 8.54 * 10 -3 g / (U·h).

Embodiment 3

[0088] The preparation of embodiment 3 L-sodium malate

[0089] In this embodiment, in addition to adopting calcium bicarbonate to prepare calcium fumarate suspension at the beginning of the method and adopting the recombinant escherichia coli introduced and expressing the fuma gene as fumarate hydratase, prepare according to the method described in Example 1 This results in sodium L-malate, wherein the calcium carbonate as a solid precipitate is recovered and recycled in the preparation of the calcium fumarate suspension.

[0090] The average productive rate of the L-sodium malate of this embodiment is 28.45g / (L h), and the catalytic efficiency of fumaric acid hydratase is 8.56 * 10 -3 g / (U·h).

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Abstract

The invention discloses a method for producing L-sodium malate through direct enzymatic conversion. The method comprises the following steps: with fumaric acid and calcium salt as the raw materials, L-calcium malate is produced under the catalysis of fumarate hydratase, then after neutralizing of sodium carbonate, L-sodium malate is prepared. According to the method, by selectively cloning and expressing the coding gene segments of the fumarate hydratase catalyzing the forward reaction, the balance of the reaction deviates towards the direction of synthesis of L-malate, meanwhile, a calcium type substrate is adopted for carrying out catalyzing, thus the problem of the product quality caused by substrate residue is solved, moreover, calcium carbonate generated as the by-product is recycled,so that no discharge of by-products exists during the whole production process, the economical conversion of atoms is realized, and thus the method has the obvious industrial application value and environmental benefit.

Description

technical field [0001] The invention relates to an enzymatic conversion production method of L-sodium malate, belonging to the field of biocatalytic preparation of organic acids and salts thereof. Background technique [0002] L-sodium malate is an optical isomer of DL-sodium malate. Compared with DL-sodium malate, L-sodium malate can directly enter the tricarboxylic acid cycle to participate in the metabolism of the human body, and has important physiological activities Features. In DL-sodium malate, the D-sodium malate contained in it is not physiologically active and can only be utilized by the body after the action of racemase in the liver and oxidoreductase in the kidney. In view of this, DL-apple Sodium malate has been banned from being used in food for the elderly, patients with liver and kidney dysfunction, and infants and young children, so as to avoid the residue of D-sodium malate in the human body and thereby reduce the metabolic burden of the human body. There...

Claims

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Application Information

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IPC IPC(8): C12P7/46
CPCC12P7/46
Inventor 周铁成王云山王自强冯晓东赵晶
Owner THE GREAT WALL BIO CHEM ENG CO LTD
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