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Immobilized modified threonine transaldolase and application thereof

A technology for immobilizing enzymes and threonine, applied in the direction of immobilizing enzymes, transferases, oxidoreductases, etc., can solve the problems of high cost and large amount of enzymes

Pending Publication Date: 2022-05-27
湖南引航生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the amount of enzymes used in this method is still large and the cost is high

Method used

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  • Immobilized modified threonine transaldolase and application thereof
  • Immobilized modified threonine transaldolase and application thereof
  • Immobilized modified threonine transaldolase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Preparation of immobilized enzyme catalyst

[0037] Unless otherwise specified, the experimental methods used in the present invention are all conventional methods.

[0038] i) Reagents and instruments:

[0039] p-Methylsulfonylbenzaldehyde was purchased from Aladdin, item number M185093, purity 98%;

[0040] L-threonine was purchased from McLean, product number C10393311, analytically pure;

[0041] Pyridoxal phosphate was purchased from Aladdin, item number P136795, purity ≥98%;

[0042] Magnesium chloride was purchased from Aladdin, article number A2006034, analytically pure;

[0043] Oxidized nicotinamide adenine dinucleotide (NAD) was purchased from Aladdin, catalog number N196974, with a purity of 95%.

[0044] LX-1000NH was purchased from Lanxiao Technology.

[0045] ii) Vector and strain: the used expression vector was pET-30a(+), the plasmid was purchased from Novagen Company, and the used host cell was Escherichia coli BL21 (DE3), purchased fro...

Embodiment 2

[0060] Example 2: Preparation of immobilized enzyme (2S,3R)-p-methylsulfonylphenylserine

[0061] p-methylsulfonylbenzaldehyde 50g / L, L-threonine 42g / L, magnesium chloride 1g / L, pyridoxal phosphate 0.15g / L, co-substrate (see Table 2 for specific amounts), 100mM phosphate buffer (pH7 0) be heated to 30 ℃ of magnetic stirring uniformly, add the immobilized enzyme prepared in Example 1, start stirring reaction, react 24 hours sampling HPLC detection conversion rate (for the first time), apply mechanically after 30 times and detect conversion rate (results are shown in Table 3) .

[0062]

[0063] table 3

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PUM

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Abstract

The invention provides immobilized modified threonine transaldolase and application of the immobilized modified threonine transaldolase in catalytic synthesis of 3-phenyl-L-serine and derivatives thereof. Compared with a free enzyme, the immobilized enzyme has the advantages that the purity of the product is improved, and the use cost of the enzyme and the purification cost of subsequent reaction are reduced.

Description

technical field [0001] The present invention relates to the field of enzyme engineering and immobilized enzymes. Specifically, the present invention relates to the application of immobilized modified threonine transaldolase to catalyze the synthesis of 3-phenyl-L-serine and its derivatives. Background technique [0002] 3-Phenyl-L-serine and its derivatives have the following general structural formula: [0003] [0004] 3-Phenyl-L-serine and its derivatives are important intermediates in organic synthesis, which are widely used in drug synthesis. For example: (2S,3R)-p-methylsulfonyl phenylserine is a key intermediate in the synthesis of veterinary drugs florfenicol and thiamphenicol, and (2S,3R)-p-nitrophenylserine can be used as the antibacterial drug chloramphenicol. key intermediates. In addition, 3-phenyl-L-serine and its derivatives can also be prepared by simple transformation to chiral aziridines (David Tanner, Chiral Aziridines-Their Synthesis and Use in Ster...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/091C12P13/04C12N9/02C12N9/04C12N9/10
CPCC12N11/091C12N9/1014C12N9/0008C12N9/0006C12P13/04C12Y102/01002C12Y201/02001C12Y101/01047C12Y101/01002Y02P20/584
Inventor 谢新开梁晓亮杜好勉李林林
Owner 湖南引航生物科技有限公司
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