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A method of microbial transformation

A technology of microorganisms and transformation processes, applied in the field of industrial microorganisms, can solve the problem of high price

Active Publication Date: 2019-07-23
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 phenylpyruvate is a more direct substrate, its price is higher, so L-phenylalanine is the best substrate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Analytical Determination of Transformation Products.

[0025] 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 100 bottles, sterilized at 120°C for 20 minutes. Take Providencia rustigianii ATCC 33673 glycerol tube seed solution 1mL to inoculate, the fermentation temperature is 30°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. Get 40g wet thallus and put into the 1000ml solution that L-phenylalanine concentration is 20g / L, mix well. Shake (100 rpm) at 37°C for 48 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...

Embodiment 2

[0029] 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.

[0030]Inoculate 1 bottle with 1 mL of Providencia rettgeri ATCC 29944 glycerol tube seed solution, incubate in an anaerobic incubator at 35°C for 72 hours, and wash the bacterial cells obtained by centrifugation twice with sterile water. Take 0.1g of wet bacteria and put it into 4ml solution with L-phenylalanine concentration of 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....

Embodiment 3

[0033] 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 Providencia rettgeri CICC 10488 glycerol tube seed solution 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-phenylalanine 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 liquid was filtered through a microporous membrane, and the concentration of D-phenyll...

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Abstract

The invention relates to a method for producing R-(+)-2-hydroxy-3-phenylpropanoic acid by transforming L-phenylalanine by adopting providencia microbes. The method is simple in process and has important industrial application value.

Description

technical field [0001] The invention adopts Providencia bacteria to transform L-phenylalanine to produce D-phenyllactic acid, belonging to the field of industrial microorganisms. Background technique [0002] D-phenyllactic acid, scientific name R-(+)-2-hydroxy-3-phenylpropanoic acid, English name: R-(+)-2-hydroxy-3-phenylpropanoic acid, D-(+)-3- Phenyllactic acid. [0003] Phenyllactic acid is a new type of biological preservative discovered in recent years that can be secreted by some lactic acid bacteria. It has two enantiomers of D-phenyllactic acid and L-phenyllactic acid, which can effectively inhibit a variety of bacteria that cause food spoilage and fungi that produce toxins. Studies have shown that the antibacterial ability of D-phenyllactic acid is slightly higher than that of L-phenyllactic acid. [0004] As a new biological preservative, phenyllactic acid has important application value in the food industry. At present, the production of phenylpyruvate is mai...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/42C12R1/01
CPCC12P7/42
Inventor 蔡宇杰沈天成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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