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Pseudomonas putida strain and application thereof in preparing glycolic acid

A technology of Pseudomonas putida and Pseudomonas putida is applied in the field of enzymatic preparation of glycolic acid, which can solve the problem of no human glycolic acid and the like, and achieve the effects of large economic value, low price and high product purity

Inactive Publication Date: 2012-07-04
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Haloacetic acid dehalogenase is currently mainly used for the environmental treatment of chloroacetic acid in sewage or industrial wastewater, and so far no one has used it to produce glycolic acid

Method used

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  • Pseudomonas putida strain and application thereof in preparing glycolic acid
  • Pseudomonas putida strain and application thereof in preparing glycolic acid
  • Pseudomonas putida strain and application thereof in preparing glycolic acid

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Experimental program
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Embodiment Construction

[0033] The enzyme activity unit used in the present invention is defined as: the amount of enzyme needed to hydrolyze chloroacetic acid to produce 1 μmol glycolic acid per minute.

[0034] The composition of the seed medium used in the present invention: 3.2 g of disodium hydrogen phosphate dodecahydrate, 1.5 g of potassium dihydrogen phosphate, 0.098-0.2 g of magnesium sulfate, 0.2-0.5 g of yeast extract, 0.5-1.0 g ammonium sulfate, add water to 1L, and sterilize.

[0035] The composition of the fermentation medium used in the present invention is as follows: on the basis of the seed medium, an inducer sterilized by filtration is added, or an auxiliary carbon source is further added to increase enzyme production.

[0036] For the analytical methods of chloroacetic acid and glycolic acid, see the appendix figure 1 .

[0037] (1) Strain screening and identification

[0038] In the present invention, 8 soils are obtained from the soil near the garbage dump, the soil near many...

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Abstract

The invention discloses a pseudomonas putida strain and an application thereof in preparing glycolic acid. The strain is named as pseudomonas putida strain CA15 and can be used for preparing the glycolic acid. The method for using the pseudomonas putida strain CA15 to prepare the glycolic acid mainly comprises the following steps: adding an inducing agent to carry out induction culture on the pseudomonas putida strain CA15, obtaining cells by centrifugation or extracting crude enzyme further by breaking the cells after obtaining the cells; and using the obtained resting cells or the crude enzyme solution as a catalyst, adding chloroacetic acid salt in buffer solution as primer for reaction, and then collecting the glycolic acid from the reaction liquid. The conversion ratio of the glycolic acid obtained by the invention is more than 99 percent and the purity thereof is more than 99 percent. The method is obviously characterized by mild reaction condition, no pollution, simple technique and the like.

Description

technical field [0001] The invention relates to a Pseudomonas putida strain and an enzymatic method for preparing glycolic acid. Background technique: [0002] Glycolic acid (glycolic acid) is an important intermediate, widely used in daily necessities, industrial cleaning agents, polymer monomers and pharmaceutical fields. The traditional method of preparing glycolic acid is through chemical synthesis, but usually chemical synthesis requires high temperature and high pressure, and the purity is difficult to meet the demand. At present, the relatively mature chemical method for preparing glycolic acid mainly includes the alkaline hydrolysis of chloroacetic acid. However, the disadvantages of this method are that the reaction efficiency is low, and the product is easy to produce polyester or lactide and the like. Biological preparation of glycolic acid has received widespread attention, mainly including nitrilase hydrolysis of glycolic acid and asymmetric oxidation of ethyl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P7/42C12R1/40
Inventor 林春娇杨立荣吴坚平徐刚
Owner ZHEJIANG UNIV
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