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Production of alpha-keto butyrate

A ketobutyric acid, microorganism technology, applied in the directions of microorganism-based methods, microorganisms, biochemical equipment and methods, etc., can solve the problem of difficult to obtain α-ketobutyric acid accumulation and other problems

Inactive Publication Date: 2005-03-09
SOC DES PROD NESTLE SA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In conclusion, accumulation of α-ketobutyrate may be difficult to obtain by biotransformation

Method used

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  • Production of alpha-keto butyrate
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Example 1: Accumulation of α-ketobutyric acid by biotransformation of threonine in Neurospora crassa

[0090] Materials and methods

[0091] Spores of Neurospora crassa strain with ilv-3(-) genotype were purchased from Kansas Medical Center, catalog number FGSC 575, conjugation type A, and spread on agar in Erlenmeyer flasks for the preparation of the starting material. preparation. The composition of the "Horowitz" medium used for this purpose is given in Table 1 below.

[0092] Element

amount per liter

Total Salt Solution*

200ml

Glycerin (87%)

185ml

casein hydrolyzate

0.5g

Yeast extract

5g

malt extract

5g

agar

20g

Biotin solution**

100μl

sterile water

Adjust to 1000ml

pH

6.5

[0093] * total salt solution

[0094] 25g K-Tartrat (dipotassium tartrate hemihydrate, Merck)

[0095] 20g sodium nitrate

[0096] 5gKH 2 PO 4

[0097] 2.5gMgSO 4 ×7...

Embodiment 2

[0120] Example 2: Two-step fermentation method increases the accumulation of α-ketobutyric acid

[0121] Materials and methods

[0122] The same strains, media and equipment as cited in Example 1 were used.

[0123]In the first fermentation, biomass production was achieved by inoculating a medium containing, according to the data in Example 1, fortified Vogel-salt solution, L-valine and L-isoleucine (added later threonine). The pH was 5.8 at the beginning of the fermentation, and dropped to 4.4 after 72 hours with gentle shaking (200 rpm) at 26°C.

[0124] The mycelial cells were then filtered (0.22 μm) and transferred to a different medium, phosphate buffered saline (0.1 M ). The pH was adjusted to 10 with NaOH (10N).

[0125] After a total of 110 hours of reaction at 25°C and 200 rpm, biotransformation was interrupted by filtering the medium to remove biomass and the filtrate was stored at 4°C for further analysis.

[0126] result

[0127] A high content of α-ketobuty...

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PUM

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Abstract

A process of obtaining alpha-keto butyrate by bioconversion is disclosed. The molecule is accumulated in the medium and is apt to subsequent processing to food ingredients, such as top-note flavors. A suitable strain is a natural ilv-3 mutant Neurospora crassa strain, which is capable of accumulating alpha-keto butyrate in the fermentation medium. By adjusting specific medium parameters, up to 8 g / l medium of alpha-keto butyrate can be accumulated.

Description

[0001] The present invention relates to a method for the accumulation of alpha-ketobutyric acid. The invention also relates to the use of natural mutant microorganisms and / or extracted enzymes for the production of natural alpha-ketobutyric acid. The invention also relates to methods for obtaining emoxyfurone. Background technique [0002] α-ketobutyric acid is a synonym of 2-oxobutyric acid, which can be accumulated by a variety of microorganisms. However, according to reports to the contrary, it has also been shown that this molecule is produced in an equilibrium concentration range and is therefore not suitable for accumulation. [0003] In JP03183476, microorganisms belonging to the genus Pseudomonas are known to be able to convert 2,3-dihydroxybutyrate into α-ketobutyrate. However, Pseudomonas is not a food-grade microorganism, so α-ketobutyric acid obtained by fermentation of this microorganism cannot be easily applied to food. The same applies to derivatives obtained...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/40C12P17/04C12R1/645
CPCC12P17/04C12P7/40
Inventor B·D·祖尔布里根N·雷克黑夫M·梅尔曼-德-凯姆普斯K·勒奇
Owner SOC DES PROD NESTLE SA
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