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L-lactic acid catalysis reaction system and preparation method of L-lactic acid

A technology of catalytic reaction and reaction system, applied in the field of enzyme catalysis production of L-lactic acid, can solve the problems of inconvenient industrial production and expensive substrates, etc.

Active Publication Date: 2018-02-06
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The chloropropionic acid enzymatic method uses DL-2-chloropropionic acid as a substrate to produce L or D-lactic acid. The substrate of this method is more expensive and not easy for industrial production

Method used

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  • L-lactic acid catalysis reaction system and preparation method of L-lactic acid
  • L-lactic acid catalysis reaction system and preparation method of L-lactic acid
  • L-lactic acid catalysis reaction system and preparation method of L-lactic acid

Examples

Experimental program
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Effect test

experiment example 1

[0039] Experimental example 1. In vitro multi-enzyme catalyzed conversion of glucose into L-lactic acid

[0040] Combination of glucose dehydrogenase, dihydroxy-acid dehydratase, 2-keto-3-deoxygluconate aldolase, glyceraldehyde dehydrogenase, dihydroxy-acid dehydratase and L-lactate dehydrogenase to establish L-lactate catalyzed reaction system.

[0041] Glucose is converted into L-lactic acid by the L-lactic acid catalytic reaction system ( figure 1), including: (1) glucose dehydrogenase (GDH, EC 1.1.1.47), which catalyzes the production of gluconic acid from glucose; (2) dihydroxyacid dehydratase (DHAD, EC 4.2.1.9), which catalyzes the production of 2-ketone from gluconic acid -3-deoxygluconic acid (3) 2-keto-3-deoxygluconic acid aldolase (KDGA, EC4.1.2.14), catalyzes 2-keto-3-deoxygluconic acid to generate glyceraldehyde and pyruvate; (4 ) Glyceraldehyde dehydrogenase (ALDH, EC1.2.1.3), which catalyzes glyceraldehyde to generate glyceric acid; (5) dihydroxy acid dehydrata...

experiment example 2

[0049] Experimental example 2. In vitro multiple enzymes catalyze the conversion of glucose into L-lactic acid (determination of enzyme concentration)

[0050] In a 1mL reaction system containing 100mM HEPES buffer (pH7.0), 5mM divalent magnesium ions, 5mM NAD + , 27.75mM glucose, the catalytic reaction was carried out at 50°C, and the reaction time was 12 hours. The proportions of different enzymes were changed respectively to finally determine the proportions of enzymes required in the reaction system.

[0051] After the reaction, if Figure 5 shown. First, optimize the amount of glucose dehydrogenase added. Only the added amount of glucose dehydrogenase is changed, and the added amount of other components in the reaction system does not change. After the reaction, the concentration of L-lactic acid is determined to determine the The optimal addition amount of hydrogenase was determined to be 1U / mL. After the addition amount of glucose dehydrogenase is determined, the ad...

experiment example 3

[0052] Experimental example 3. In vitro multi-enzyme catalyzed conversion of glucose into L-lactic acid (determination of coenzyme concentration)

[0053] After determining the amount of enzyme to be added in the reaction system, carry out coenzyme NAD + concentration optimization.

[0054] In a 1mL reaction system containing 100mM HEPES buffer (pH7.0), 5mM divalent magnesium ions, 27.75mM glucose, 1U / mL glucose dehydrogenase, 2U / mL dihydroxy acid dehydratase, 1U / mL 2-keto-3-deoxygluconate aldolase, 1U / mL glyceraldehyde dehydrogenase, 1U / mL L-lactate dehydrogenase, catalyze the reaction at 50°C, and the reaction time is 12 hours . Altered coenzyme NAD + The addition ratio is 0-10mM, and finally determine the addition ratio of the required coenzyme in the reaction system.

[0055] After the reaction, by Image 6 It can be seen that when the coenzyme NAD + The production rate of lactic acid did not increase significantly after the addition amount of 5mM. Taking into cons...

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Abstract

The invention provides an L-lactic acid catalysis reaction system and a preparation method of L-lactic acid. The L-lactic acid catalysis reaction system contains glucose dehydrogenase, dihydroxyacid anhydrase, 2-keto-3-deoxygluconate, glyceraldehyde phosphate dehydrogenase and L-lactic dehydrogenase. The L-lactic acid can be obtained without adding ATP (Adenosine Triphosphate) by performing a cascade enzymatic catalytic reaction in a reactor by utilizing five enzymes of the reaction system and taking glucose as a raw material.

Description

technical field [0001] The invention relates to the field of enzyme-catalyzed production of L-lactic acid, in particular to an L-lactic acid catalytic reaction system and a preparation method of L-lactic acid. Background technique [0002] Lactic acid, also known as α-hydroxypropionic acid, is divided into D-lactic acid and L-lactic acid because it contains an asymmetric carbon atom. Lactic acid has good applications in food, medicine, biodegradable plastics and cosmetics industries. The application of lactic acid in the food industry is mainly as a sour agent, and it is used as a bacteriostat and flavor regulator in the brewing industry. Lactic acid can be directly formulated into medicine in the pharmaceutical industry, and lactate can be used as a disinfectant. Because L-lactic acid contains hydroxyl and carboxyl groups, it has automatic esterification ability, dehydration polymerization into poly L-lactic acid, highly optically isomerized poly L-lactic acid is used in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/04C12N9/02C12N9/88C12P7/56
CPCC12N9/0006C12N9/0008C12N9/88C12P7/56C12Y101/01027C12Y101/9901C12Y102/01C12Y401/02C12Y402/01009
Inventor 游淳徐淑霞谢雷朋张以恒
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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