Method for preparing alpha-ketobutyric acid by using L-threonine as substrate

A technology of ketobutyric acid and threonine, which is applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., achieves the effects of high conversion rate, convenient operation, and beneficial to subsequent separation and extraction.

Active Publication Date: 2012-05-02
上海肆芃科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows creating alpha - ketone compounds from lysophospholic acids (LPA). These processes involve converting certain chemicals called phytases into different types of amino acids that are important building blocks for various products such as foodstuffs and medicines. By selecting specific bacteria like Microbacillus sp., this process produces these molecules efficiently while also producing other valuable materials through their metabolism.

Problems solved by technology

This patents discuss various methods used to produce alpha -lyservalero hydroxyl succinate(α-AS), which can be useful intermediates or precursors therapy agents such as beta thymosins produced through enzymatic reactions with other molecules like lysosergene lacticum nucleate hydrocinnoline monofunctional alkaloids called pseudoplasmideutrosporone dibasic esters. These techniques involve modifying certain chemical structures within these compounds without changing their structure themselves.

Method used

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  • Method for preparing alpha-ketobutyric acid by using L-threonine as substrate
  • Method for preparing alpha-ketobutyric acid by using L-threonine as substrate
  • Method for preparing alpha-ketobutyric acid by using L-threonine as substrate

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Experimental program
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Effect test

Embodiment 2

[0061] (1) Preparation of suspension containing biocatalyst

[0062] Bacillus subtilis ATCC23857 was selected and aerobically cultured in LB medium in a conventional shake flask or fermenter; the bacteria were separated and collected, washed with pH 7.4 potassium phosphate buffer for 3 times, and the isolated complete microbial cells were obtained as Bio-catalyst. The biocatalyst is resuspended in potassium phosphate buffer or deionized water, so that the concentration of the biocatalyst reaches 200 g wet cells / liter, and the suspension containing the biocatalyst is obtained, and stored at 4°C for use;

[0063] (2) Conversion

[0064] The biocatalyst-containing suspension prepared in step (1) is mixed with the L-threonine aqueous solution, and the concentration of L-threonine in the mixture is 10 grams per liter, and the biocatalyst concentration is 60 grams of wet cells / liter; under the conditions of 30°C and pH 8.0, the reaction was shaken at 200 rpm for 24 hours to obtai...

Embodiment 3

[0070] (1) Preparation of suspension containing biocatalyst

[0071] Select Pseudomonas stutzeri (P.stutzeri) SDM CCTCC NO: M206010, aerobic culture in LB medium with conventional shake flask or fermenter; separate and collect the bacteria, wash the bacteria with pH 7.4 potassium phosphate buffer 3 times, the isolated complete cells of microorganisms are biocatalysts. The biocatalyst is resuspended in potassium phosphate buffer or deionized water, so that the concentration of the biocatalyst reaches 200 g wet cells / liter, and the suspension containing the biocatalyst is obtained, and stored at 4°C for use;

[0072] (2) Conversion

[0073] The biocatalyst-containing suspension prepared in step (1) is mixed with the L-threonine aqueous solution, and the concentration of L-threonine in the mixture is 60 grams per liter, and the biocatalyst concentration is 120 grams of wet cells / liter; under the conditions of 65°C and pH 7.0, the reaction was shaken at 200 rpm for 24 hours to ...

Embodiment 4

[0079](1) Preparation of suspension containing biocatalyst

[0080] Corynebacterium glutamicum ATCC13032 was selected and aerobically cultured in LB medium with conventional shake flasks or fermenters; the bacteria were separated and collected, washed with pH 7.4 potassium phosphate buffer for 3 times, and the obtained microbial complete cells were isolated is a biocatalyst. The biocatalyst is resuspended in potassium phosphate buffer or deionized water, so that the concentration of the biocatalyst reaches 200 g wet cells / liter, and the suspension containing the biocatalyst is obtained, and stored at 4°C for use;

[0081] (2) Conversion

[0082] The biocatalyst-containing suspension prepared in step (1) is mixed with the L-threonine aqueous solution, and the concentration of L-threonine in the mixture is 20 grams per liter, and the biocatalyst concentration is 40 grams of wet cells / liter; under the conditions of 60°C and pH 10.0, the reaction was shaken at 200 rpm for 20 ho...

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Abstract

The invention discloses a method for preparing alpha-ketobutyric acid by using L-threonine as a substrate, wherein a product, alpha-ketobutyric acid is prepared by using microbial complete cells containing synthetic L-threonine dehydratase as a biological catalyst, using the L-threonine as the substrate, and performing oscillating transformation at a rate of 50 to 250 turns/minute under the conditions that the temperature is between 20 and 65 DEG C and the pH is between 7.0 and 11.0. The method has the following characteristics that (1) a biological catalysis method is used, a reaction system is simple, reaction conditions are mild, steps are easy, and the operation is simple; and the biological catalyst can be easily removed, and the separation and purification of subsequent products are facilitated; (2) the transformation rate of the alpha-ketobutyric acid generated by the substrate, L-threonine can reach more than 99.6 percent, and the product, alpha-ketobutyric acid can be accumulated to reach a higher concentration; and (3) the substrate, L-threonine has a low price and is easy to obtain. According to the method, a basis of realizing high-efficient production of the alpha-ketobutyric acid is established.

Description

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Claims

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Application Information

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Owner 上海肆芃科技有限公司
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