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Process for preparing pyruvic acid using lactic acid oxidase or whole cell transformed lactic acid contg. said oxidase

A technology of lactic acid oxidase and complete cells, which is applied in the field of biological transformation of sodium lactate to prepare pyruvate, can solve the problems of expensive substrate and high cost, and achieve the effect of low substrate cost, low cost and convenient operation

Inactive Publication Date: 2006-10-18
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the substrate of this method is more expensive and the cost is higher
[0012] Utilize the lactic acid oxidase (LOD) of Acinetobacter sp. (Acinetobacter sp.) or Pseudomonas sp. (Pseudomonas sp.) or other microorganisms or complete cell to carry out the method for converting sodium lactate to prepare pyruvate (do not produce peroxidation in the reaction system) Hydrogen), no report

Method used

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  • Process for preparing pyruvic acid using lactic acid oxidase or whole cell transformed lactic acid contg. said oxidase
  • Process for preparing pyruvic acid using lactic acid oxidase or whole cell transformed lactic acid contg. said oxidase

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Embodiment 1: Utilize the lactic acid oxidase crude enzyme liquid of Acinetobacter to convert DL-sodium lactate to prepare the method for pyruvate

[0047] (1) Strain selection: Acinetobacter sp. ATCC11171 was selected.

[0048] (2) Slant culture: inoculate the above strains on a solid slant medium containing 1.8% agarose and added with 0.5% DL or L-sodium lactate, and culture the bacteria at 25°C for 20 hours.

[0049] (3) First-level seed culture: put the strain cultivated in step (2) into 50 mL of BLM containing 0.5% DL or L-sodium lactate (using a 300 mL Erlenmeyer shaker flask) with an inoculation loop under sterile conditions, at 25°C Under the conditions, shake culture on a shaker for 20 hours to obtain first-class seeds.

[0050] (4) Expansion culture: inoculum amount of 5% volume / volume percentage, connect first-level seeds in 300mL BLM containing 0.5% DL or L-sodium lactate (use 1L Erlenmeyer shaker flask), shake on a shaker at 25°C Cultivate for 15 hours to...

Embodiment 2

[0057] Embodiment 2: the method for preparing pyruvate by transforming DL-sodium lactate with whole cells of Pseudomonas

[0058] (1) Strain selection: Pseudomonas sp. ATCC11452 was selected.

[0059] (2) Slant culture: inoculate the above strains on a solid slant medium containing 2.0% agarose and 2% DL or L-sodium lactate, and culture the bacteria at 37°C for 30 hours.

[0060] (3) First-level seed culture: put the strain cultivated in step (2) into 100 mL of BLM containing 2% DL or L-sodium lactate (using a 500 mL Erlenmeyer flask) with an inoculation loop under sterile conditions, at 37°C Next, shake culture on a shaker for 30 hours to obtain primary seeds.

[0061] (4) Expansion culture: inoculum amount of 5% volume / volume percentage, connect first-level seeds in 600mL BLM containing 2% DL or L-sodium lactate (using a 1L Erlenmeyer flask), and culture on a shaker at 37°C After 10 hours, secondary seeds were obtained.

[0062] (5) Fermentation tank culture: with 5% volu...

Embodiment 3

[0067] Embodiment 3: Utilize the method for preparing pyruvate by transforming DL-sodium lactate with complete cells of Acinetobacter

[0068] (1) Strain selection: Acinetobacter sp. ATCC11171 was selected.

[0069] (2) Slant culture: the above strains were inoculated on a solid slant medium containing 1.5% agarose and 1.0% DL or L-sodium lactate, and cultured at 30° C. for 28 hours.

[0070] (3) First-level seed culture: put the strain cultivated in step (2) into 25 mL of BLM containing 1.0% DL or L-sodium lactate (using a 300 mL Erlenmeyer flask) with an inoculation loop under sterile conditions, at 30°C Next, shake culture on a shaker for 25 hours to obtain primary seeds.

[0071] (4) Expansion culture: Inoculate with 5% volume / volume percentage, place the first-stage seeds in 500 mL of BLM containing 1.0% DL or L-sodium lactate (using a 2L Erlenmeyer shaker flask), shake on a shaker at 30°C Cultivate for 30 hours to obtain secondary seeds.

[0072] (5) Fermentation tank...

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Abstract

The invention discloses a method for converting DL or L-sodium lactate into pyruvate by utilizing lactate oxidase or complete cells containing the enzyme, the method comprising (1) strain selection; (2) slant culture; (3) seed culture (4) fermenter culture; (5) collection of bacteria; (6) transformation experiment; (7) steps such as separating samples. The invention has the characteristics of simple method, low cost, high conversion efficiency, simple method for extracting products, etc., and has great application prospects in the industrial production of pyruvic acid.

Description

(1) Technical field [0001] The invention relates to a method for preparing pyruvate by converting sodium lactate through a biological method, in particular to a method for preparing pyruvate by using lactate oxidase (LOD) or a complete cell containing the enzyme to convert sodium lactate. (2) Background technology [0002] Pyruvate not only plays a very important role in energy metabolism, but also is the precursor of many useful compounds. Therefore, it has a wide range of uses in industrial and scientific research such as chemical, pharmaceutical and agricultural chemicals. The preparation method of pyruvic acid has therefore become a research hotspot. [0003] Until the 1990s, the industrial production of pyruvic acid followed the tartaric acid dehydration and decarboxylation method published by Howard and Fraser in "Org Synth Coll Vol1: 475-480 Preparation of pyruvic acid" in 1932. The disadvantages are (1) low yield of pyruvic acid (0.29-0.30 g / g for tartaric acid), an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/40C12R1/01
Inventor 许平马翠卿魏中浩
Owner SHANDONG UNIV