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A kind of preparation method of L-serine

A technology of serine and glycine, which is applied in the field of L-serine preparation, can solve the problems of reducing enzyme activity, adverse effects of enzyme activity, and unsatisfactory L-serine yield, and achieves the effect of increasing yield

Active Publication Date: 2021-06-01
WUHAN POLYTECHNIC UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, in the existing methods for preparing L-serine by enzymatic conversion, chemical reagents such as CTAB (hexadecyltrimethylammonium bromide) are generally used for cell disruption. Substances remain in the mixture after cell breakdown, and the existence of such chemical substances will adversely affect the activity of the enzyme in the enzymatic reaction, reduce the activity of the enzyme, and lead to an unsatisfactory yield of L-serine

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  • A kind of preparation method of L-serine

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preparation example Construction

[0033] In view of this, the present invention proposes a kind of preparation method of L-serine, by choosing the mode of physical method to break the cell, in order to avoid the problem of reducing the yield of L-serine brought when using chemical reagents to break the cell, figure 1 Shown is an example of the preparation method of L-serine provided by the present invention. see figure 1 , in this embodiment, the preparation method of said L-serine comprises the following steps:

[0034] Step S10, using a physical method to disrupt the cells of the bacteria to obtain cell disruptions;

[0035] In this embodiment, the physical method of disrupting cells can be carried out by using, for example, a high-pressure cell disruptor. In specific operations, it is generally necessary to prepare the bacteria into a solution, and then use a cell disruptor to process the cells under a certain pressure and flow rate. The crushing can be specifically carried out according to the following ...

Embodiment 1

[0048] (1) Suspend 0.2 kg of bacteria in 3 L Tris-HCl buffer solution with a pH value of 7.0, and use APV-2000 high-pressure cell disruptor to disrupt the cells at a pressure of 600 bar and a flow rate of 8 L / h to obtain cell disruption thing;

[0049] (2) Place the crushed cells in a pressurized incubator, first pass in air to pressurize to 6bar, incubate for 5h, then open the valve to release the pressure to normal pressure, then pass in nitrogen to pressurize to 0.1bar, and incubate for 30min to obtain incubation mixture;

[0050] (3) Transfer the incubation mixture into an enzymatic reaction kettle, and add glycine, pyridoxal 5-phosphate, tetrahydrofolic acid and formaldehyde to prepare an enzymatic reaction system, wherein glycine, pyridoxal 5-phosphate, tetrahydrofolate Corresponding to the concentration of formaldehyde is 2.4mol / L, 0.3mmol / L, 4mmol / L and 10mmol / L, and the pH value of the enzymatic reaction system is adjusted to 7.0; during the enzymatic reaction, maint...

Embodiment 2

[0053] (1) Suspend 0.2 kg of bacteria in 3 L Tris-HCl buffer solution with a pH value of 6.0, and use APV-2000 high-pressure cell disruptor to disrupt the cells at a pressure of 500 bar and a flow rate of 6 L / h to obtain cell disruption thing;

[0054] (2) Place the crushed cells in a pressurized incubation kettle, first pass in air to pressurize to 4bar, incubate for 6h, then open the valve to release the pressure to normal pressure, then pass in nitrogen to pressurize to 0.05bar, incubate for 35min, and then incubate mixture;

[0055] (3) Transfer the incubation mixture into an enzymatic reaction kettle, and add glycine, pyridoxal 5-phosphate, tetrahydrofolic acid and formaldehyde to prepare an enzymatic reaction system, wherein glycine, pyridoxal 5-phosphate, tetrahydrofolate Corresponding to the concentration of formaldehyde is 2mol / L, 0.2mmol / L, 3mmol / L and 8mmol / L, and the pH value of regulating the enzymatic reaction system is 6.0; Nitrogen positive pressure, under th...

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Abstract

The invention discloses a preparation method of L-serine, which comprises the following steps: using a physical method to crush the cells of the bacterium to obtain a cell crush; incubating the cell crush to obtain an incubation mixture; Glycine, pyridoxal 5-phosphate, tetrahydrofolate and formaldehyde are added into the mixture, and an enzymatic reaction is carried out under a nitrogen atmosphere until the enzymatic reaction balance is reached to obtain L-serine. The present invention uses a physical method to crush the cells of the bacterium, and then pressurizes the cell crushed matter, and then uses the incubation mixture obtained from the pressurized incubation as a raw material to perform an enzymatic reaction under a nitrogen atmosphere to prepare L-serine, which is effective Improved yield of L‑serine by enzymatic conversion.

Description

technical field [0001] The invention relates to the technical field of L-serine preparation, in particular to a preparation method of L-serine. Background technique [0002] L-serine is in many metabolic pathways of the body and plays important physiological functions in the body. At the same time, as a raw material, L-serine is widely used in industries such as chemical industry, pharmaceuticals, food, cosmetics and biological pesticides, so its demand is increasing day by day, and it is one of the most expensive amino acids on the market. [0003] Serine hydroxymethyltransferase is the key enzyme for the preparation of L-serine by in vitro enzymatic method. It is a pyridoxalase with pyridoxal 5-phosphate (PLP) as a coenzyme, and can be produced in the presence of tetrahydrofolate (THF). It catalyzes the condensation of formaldehyde and glycine to generate L-serine. In large-scale production, how to increase the yield of L-serine has always been considered as an effective...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/06
CPCC12P13/06
Inventor 梅运军董文华张磊杨奕
Owner WUHAN POLYTECHNIC UNIVERSITY