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Microorganism converting method

A microbial conversion, tyrosine technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as expensive and unstable

Active Publication Date: 2016-05-25
卓虹超源生物科技(郑州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although 4-hydroxyphenylpyruvate is a more direct substrate, it is expensive and unstable, so L-tyrosine is the best substrate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Analytical Determination of Transformation Products.

[0022] Prepare medium as follows: yeast extract 5g / L, peptone 10g / L, sodium chloride 5g / L. The liquid volume in the 500ml Erlenmeyer flask is 100ml, a total of 50 bottles, sterilized at 120°C for 20 minutes. Take ProvidenciarustigianiiATCC33673 glycerol tube seed solution 1mL for inoculation, fermentation temperature is 30°C, shaker speed is 200rpm. After culturing for 24 hours, the fermentation broth was centrifuged (10,000 rpm for 10 min), the supernatant was removed to obtain bacterial cells, and the bacterial cells obtained by centrifugation were washed twice with sterile water. Take 20g of wet bacteria and put it into 10000ml of solution whose L-tyrosine concentration is 0.3g / L, and mix well. Shake (100 rpm) at 37°C for 24 hours, then heat in a water bath at 100°C for 3 minutes to kill bacteria. Then centrifuge (10000rpm, time 10min) to remove the bacteria, and obtain the transformed supernatant. Liquid pha...

Embodiment 2

[0026] Comparison of aerobic and anaerobic cultures. Preparation medium: glucose 30g / L, peptone 20g / L, diammonium hydrogen phosphate 1g / L, potassium dihydrogen phosphate 1g / L, magnesium sulfate 0.3g / L, ferrous sulfate 0.3g / L; initial pH and fermentation The pH of the process is natural. The liquid volume in the 500ml Erlenmeyer flask is 100ml, a total of 2 bottles, sterilized at 120°C for 20 minutes.

[0027] Take ProvidenciarettgeriATCC29944 glycerol tube seed solution 1mL to inoculate 1 bottle, culture in an anaerobic incubator at 35°C for 72 hours, and the bacteria obtained by centrifugation are washed twice with sterile water. Take 0.1g of wet bacteria and put it into 4ml solution with L-tyrosine concentration of 0.2g / L, mix well, put it in an anaerobic incubator at 35°C for 24 hours, then heat it in a water bath at 100°C for 3 minutes to kill the bacteria body. Then centrifuge (10000rpm, time 10min) to remove the bacteria, and obtain the transformed supernatant. Liquid...

Embodiment 3

[0030] The medium composition is: glucose 1g / L, urea 1g / L, diammonium hydrogen phosphate 1g / L, potassium dihydrogen phosphate 0.1g / L, magnesium sulfate 0.1g / L, ferrous sulfate 0.1g / L. The volume of liquid in a 250ml Erlenmeyer flask is 50ml, sterilized at 120°C for 20 minutes. Take Providenciarettgeri CICC10488 glycerol tube seed liquid 0.5mL to inoculate, the fermentation temperature is 35°C, and the shaker speed is 200rpm. After culturing for 24 hours, the fermentation broth was centrifuged (10,000 rpm for 10 min), the supernatant was removed to obtain bacterial cells, and the bacterial cells obtained by centrifugation were washed twice with sterile water. Take 0.05g of wet bacteria and put it into 2ml solution with L-tyrosine concentration of 0.1g / L, adjust the pH to 6, and mix well. Static transformation was carried out in an anaerobic incubator for 24 hours, the transformation solution was filtered through a microporous membrane, and the concentration of D-4-hydroxypheny...

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PUM

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Abstract

The invention relates to a method for producing R-(+)-3-(4-hydroxyphenyl)-2-hydracrylic acid by converting L-tyrosine with Providencia microorganisms. The method is simple in process and has significant industrial application value.

Description

technical field [0001] The invention adopts Providencia bacteria to transform L-tyrosine to produce D-4-hydroxyphenyl lactic acid, belonging to the field of industrial microorganisms. Background technique [0002] D-4-hydroxyphenyllactic acid, also known as p-hydroxyphenyllactic acid, scientific name R-(+)-3-(4-hydroxyphenyl)-2-hydroxypropionic acid, English name: D-4-hydroxyphenyllactic acid, D-p-Hydroxyphenyllactic acid, R-(+)-2-hydroxy-3-(4-hydroxyphenyl) propanoic acid. [0003] 4-Hydroxyphenyl lactic acid is another broad-spectrum, low-toxic biological preservative produced in the fermentation process of lactic acid bacteria, which is currently mainly produced by lactic acid bacteria (Chinese patent CN200810244053.2). 4-Hydroxyphenyl lactic acid is another broad-spectrum, low-toxic biological preservative produced in the fermentation process of lactic acid bacteria, which is currently mainly produced by lactic acid bacteria (Chinese patent CN200810244053.2). There are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/42C12R1/01
CPCC12P7/42
Inventor 蔡宇杰陈丽霞邓华祥陈佳君钟妮尔王静白亚军郑晓晖
Owner 卓虹超源生物科技(郑州)有限公司
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