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

A lactate oxidase and complete cell technology, applied in the field of biological conversion of lactic acid to prepare pyruvate, can solve the problems of high cost and expensive substrate, and achieve the effects of low cost, low substrate cost and simple culture medium

Inactive Publication Date: 2003-10-29
SHANDONG UNIV
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Problems solved by technology

However, the substrate of this method is more expensive and the cost is higher
[0011] After retrieval, utilize Acinetobacter sp. or Pseudomonas sp. or other microbia

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0058] (10) Sample detection: After the sample in step (9) is separated, 1 μL of sample is taken for injection, and the content of lactic acid and pyruvic acid is detected by HPLC, and the conversion rate is 94.3%. Embodiment 2: The method for preparing pyruvate by converting L-lactic acid into crude enzyme solution of Pseudomonas cells

[0059] (1) Strain selection: Pseudomonas sp. ATCC10838 was selected.

[0060] (2) Slant culture: the above strains were inoculated on a solid slant medium containing 1.5% agarose and 0.2% DL / L-sodium lactate, and cultured at 30° C. for 15 hours.

[0061] (3) First-level seed culture: put the strain cultivated in step (2) into 25 mL of BLM containing 0.2% DL / L-sodium lactate (use a 300 mL Erlenmeyer shaker flask) with an inoculation loop under sterile conditions, at 30°C Under these conditions, shake culture on a shaker for 15 hours to obtain primary seeds.

[0062] (4) Expansion culture: with 5% (volume ratio) inoculum amount, connect the f...

Embodiment 3

[0068] (10) Sample detection: After the sample in step (9) is separated, 1 μL of the sample is taken and injected into HPLC to determine the content of lactic acid and pyruvic acid, and the conversion rate is 96.4%. Embodiment 3: Utilize the method for the whole cell conversion DL-lactic acid of Pseudomonas to prepare pyruvate

[0069] (1) Strain selection: Pseudomonas sp. ATCC11452 was selected.

[0070] (2) Slant culture: inoculate the above strains on a solid slant medium containing 2.0% agarose and 2% DL / L-sodium lactate, and culture the bacteria at 37°C for 30 hours.

[0071] (3) First-level seed culture: put the strain cultivated in step (2) into 100 mL of BLM containing 2% DL / L-sodium lactate (using a 500 mL Erlenmeyer shaker flask) with an inoculation loop under sterile conditions, at 37°C Under these conditions, shake culture on a shaker for 30 hours to obtain primary seeds.

[0072] (4) Expansion culture: with 5% (volume ratio) inoculum amount, connect the first-gr...

Embodiment 4

[0077] (9) Sample detection: After the sample in step (8) is separated, 1 μL of the sample is taken and injected into HPLC to determine the content of lactic acid and pyruvic acid, and the conversion rate is 98.2%. Embodiment 4: Utilize the method for the whole cell conversion DL-lactic acid of Pseudomonas to prepare pyruvate

[0078] (1) Strain selection: Pseudomonas sp. ATCC10838 was selected.

[0079] (2) Slant culture: the above strains were inoculated on a solid slant medium containing 1.5% agarose and 3% DL / L-sodium lactate, and cultured at 45° C. for 24 hours.

[0080] (3) First-level seed culture: the bacterial strain cultivated in step (2), under sterile conditions, use an inoculation loop to inoculate 1 loop in 25mL BLM containing 0.6% DL / L-sodium lactate (using a 300mL Erlenmeyer shaker flask), 45°C Under these conditions, shake culture on a shaker for 10 hours to obtain primary seeds.

[0081] (4) Expansion culture: with 5% (volume ratio) inoculum amount, connect...

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Abstract

A process for preparing pyruvic acid by use of lactate oxidase or the complete cell containing said enzyme to transform DL/L-lactic acid includes choosing bacterial strain, slant culture, seed culture, culture in fermentor, collecting thalluses, transform experiment and separating specimen. Its advantages are simple process, low cost, and high transform efficiency.

Description

(1) Technical field [0001] The invention relates to a method for preparing pyruvate by converting lactic acid through a biological method, in particular to a method for using lactate oxidase (LOD) or a whole cell containing the enzyme to convert lactic acid to prepare pyruvate. (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 continued to use the tartrate dehydration and decarboxylation method published in the article "Org Synth Coll Vol1: 475-480 Preparation of pyruvic acid" by Howard and Fraser (Howard and Fraser) in 1932. Its main disadvantages (1) The yield of pyruvic acid is low (0.29...

Claims

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Application Information

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IPC IPC(8): C12P7/40
Inventor 许平马翠卿魏中浩
Owner SHANDONG UNIV
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