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A kind of sphingobacterium and its application and method in catalytic synthesis of l(+)-tartaric acid or its salt

A technology of sphingosine and tartaric acid, applied in the biological field, can solve the problems of low conversion rate and achieve the effect of avoiding the use

Active Publication Date: 2020-12-15
HANGZHOU BIOKING BIOCHEM ENG
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, further studies have shown that the conversion of 5-keto-D-gluconic acid to L(+)-tartaric acid is not a biological conversion, but a conversion reaction catalyzed by a rare metal, and its conversion rate is also low

Method used

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  • A kind of sphingobacterium and its application and method in catalytic synthesis of l(+)-tartaric acid or its salt
  • A kind of sphingobacterium and its application and method in catalytic synthesis of l(+)-tartaric acid or its salt

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Experimental program
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Effect test

Embodiment 1

[0020] Strain screening.

[0021] Collect the root soil of wisteria in Yuhang District, Hangzhou City, add 0.5g to 50mL of seed medium, and enrich and cultivate in a shaker at 30°C and 200rpm for 2 days. The enriched culture solution was diluted with sterile water for 10 6 Afterwards, it was spread on a solid plate medium, and incubated at a constant temperature of 30°C for 2 days. A single colony was picked and placed in a test tube containing 5 mL of seed medium, and cultured at 30 °C and 200 rpm for 16 h. Pipette 3 mL of seed culture liquid into 30 mL of fermentation medium containing 30 g / L 5-keto-D-gluconate potassium, cultivate at 30°C and 200 rpm, and regularly sample and detect the L(+)-tartaric acid content in the fermentation liquid. After 20 rounds of screening, a total of 2680 strains were selected from the soil for the above fermentation experiments, and several strains of microorganisms that can generate L(+)-tartrate from 5-keto-D-gluconate were screened, amon...

Embodiment 2

[0027] Sphingobacterium BK99 utilizes potassium 5-keto-D-gluconate to generate L(+)-potassium tartrate.

[0028] A single colony was picked from the solid medium and placed in a 250mL shake flask containing 50mL of seed medium, and shaken and cultivated in a shaker at 30°C and 200rpm for 16h to obtain a seed culture solution.

[0029] Pipette 5 mL of seed culture solution into 50 mL of fermentation medium containing different concentrations of 5-keto-D-potassium gluconate, and culture in a shaker at 30°C and 200 rpm for 10 days. After the fermentation broth was centrifuged to remove bacteria, the content of L(+)-tartaric acid in the supernatant was measured by HPLC method, and the results are shown in Table 1-5. The HPLC detection conditions are: chromatographic column Astec CLC (150 9 4.6mm), column temperature 30°C, injection volume 10 μL, mobile phase 3mM CuSO 4 Aqueous solution (pH 3.2), flow rate 1.0mL / min, detection wavelength 254nm.

[0030] Table 1 Fermentation resul...

Embodiment 3

[0041] Sphingobacterium BK99 utilizes potassium 5-keto-D-gluconate to generate L(+)-potassium tartrate.

[0042] A single colony was picked from the solid medium and placed in a 250mL shake flask containing 50mL of seed medium, and shaken and cultivated in a shaker at 30°C and 200rpm for 16h to obtain a seed culture solution.

[0043] Pipette 5 mL of seed culture solution into 50 mL of fermentation medium containing 70 g / L 5-keto-D-gluconate potassium, and culture in a shaker at 200 rpm at different temperatures for 10 days. After the fermentation broth was centrifuged to remove bacteria, the content of L(+)-tartaric acid in the supernatant was measured by HPLC, and the results are shown in Table 6.

[0044] Table 6 The influence of fermentation temperature on the fermentation of Sphingobacterium to produce L(+)-tartaric acid

[0045]

[0046] Note: The data in the above table is the concentration of L(+)-tartaric acid in the fermentation broth.

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Abstract

The invention discloses a sphingobacterium and its application and method in catalyzing the synthesis of L(+)-tartaric acid or its salt. The Sphingobacterium is preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee, the preservation number is CGMCC No. 18074, and it is named: Sphingobacterium BK99. Sphingobacterium sp. BK99 of the present invention can biosynthesize 5-keto-D-gluconic acid or its salt into L(+)-tartaric acid or its salt, avoiding the use of non-renewable fossil raw materials in the current process; On the other hand, the Sphingobacterium of the present invention replaces the reported method of using rare metals to catalyze 5-keto-D-gluconic acid to synthesize L(+)-tartaric acid, avoiding the use of rare metals.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a sphingobacterium and its application in microbial strains for catalyzing the synthesis of L(+)-tartaric acid or its salt and its application method. Background technique [0002] L(+)-tartaric acid is a natural configuration organic acid, which exists in grapes, tamarind, geranium and other higher plants, and is widely used in food, medicine, chemical industry, construction, metal and other industries. [0003] The methods for obtaining L(+)-tartaric acid mainly include extraction method, chemical synthesis resolution method, enzymatic synthesis method and sugar fermentation method. Natural tartaric acid is mainly extracted from tartar, a by-product of the winemaking process, such as Misi (response surface method to optimize the extraction process of L(+)-tartaric acid in wine sludge[J]. Food Science, 2012,33(8): 49-53) et al. extracted L(+)-tartaric acid from wine lees. Its outp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P7/46C12R1/01
CPCC12P7/46C12N1/205C12R2001/01
Inventor 黄美娟谢志鹏潘海峰孙伟荣张建国
Owner HANGZHOU BIOKING BIOCHEM ENG
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