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Method for biologically synthesizing natural aromadendrin by escherichia coli through utilizing naringenin

A technology of Escherichia coli and naringenin is applied in the field of fermentation engineering to achieve the effects of simple production process, low production cost and simple composition

Inactive Publication Date: 2017-05-10
WUXI NEWWAY FERMENTATION TECH RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no report on the production of orangenin by biofermentation using naringenin as a substrate

Method used

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  • Method for biologically synthesizing natural aromadendrin by escherichia coli through utilizing naringenin
  • Method for biologically synthesizing natural aromadendrin by escherichia coli through utilizing naringenin
  • Method for biologically synthesizing natural aromadendrin by escherichia coli through utilizing naringenin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Starting strain: the starting strain is Escherichia coli (E.coli) DHK-161114CCTCC NO:M 2016645.

[0031] Plate culture: Streak inoculate the Escherichia coli stored in the puncture tube onto the plate medium, and culture in a 30°C incubator for 15-24 hours.

[0032] Preparation of Escherichia coli CCTCC NO:M 2016645 strain bank: Pick a single colony, inoculate it into the seed medium, culture it on a shaker at 200 rpm for 13-17 hours at 30°C, absorb 0.5ml of the obtained seed culture solution and transfer it to 50% In the glycerol solution, prepare a seed glycerol tube with a final concentration of 25%, and store at -80°C.

[0033] Preparation of seed liquid: absorb 20 uL of seed glycerol tube, inoculate into seed medium, and culture at 30° C. with shaking at 200 rpm for 13 to 17 hours to obtain seed liquid.

[0034] Preparation of seed medium: peptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, prepared from tap water, pH adjusted to 7.2, sterilized by high pres...

Embodiment 2

[0040] Embodiment 2 different peptone concentrations

[0041] Starting strain: the same as in Example 1.

[0042]Preparation of seed medium: same as in Example 1.

[0043] Preparation of fermentation medium: prepare peptone 0, 2.5, 5, 7.5, 10.0 and 12.5 g / L, yeast extract 5 g / L, sodium chloride 10 g / L, tap water, adjust pH to 7.2, and extinguish with high-pressure steam at 121 °C Bacteria 20min.

[0044] Pipette 20 μL seed glycerol tube into the seed shaker flask culture medium, the liquid volume is 50ml / 250ml, culture at 30°C, 200rpm shaker for 16h, and transfer to fermentation medium with different peptone concentrations according to 5% inoculum size (50ml / 250ml), under the condition of 37°C, 200rpm shaker shake culture for 3h, cool down to 30°C, add 1.5% lactose to induce 3h, maintain 30°C, add 2.7g / L substrate naringenin in total, fermentation time After 54h, the substrate and product content were measured after the fermentation was finished, and the results were shown ...

Embodiment 3

[0046] Embodiment 3 different yeast extract concentrations

[0047] Starting strain: the same as in Example 1.

[0048] Preparation of seed medium: same as in Example 1.

[0049] Preparation of fermentation medium: peptone 10g / L, yeast extract concentrations of 0, 2.5, 5.0 and 7.5g / L, sodium chloride 10g / L, tap water, pH adjusted to 7.2, sterilized by high pressure steam at 121°C for 20min .

[0050] Pipette 20 μL of seed glycerol tube into the seed shaker flask culture medium, the liquid volume is 50ml / 250ml, and culture at 30°C, 200rpm shaker for 16h, and transfer to fermentation culture with different yeast extract concentration according to 5% inoculum size Medium (50ml / 250ml), at 37°C, 200rpm shaker culture for 3h, cooled to 30°C, added 1.5% lactose for induction for 3h, maintained at 30°C, added 2.7g / L substrate naringenin in total, The fermentation time was 54h, and the substrate and product content were measured at the end of the fermentation. The results are shown ...

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Abstract

The invention discloses a method for biologically synthesizing natural aromadendrin by escherichia coli through utilizing naringenin and belongs to the field of fermentation engineering. The method takes a gene engineering strain capable of producing the aromadendrin as a starting strain; a relatively high thalli concentration is obtained by utilizing glucose and an organic nitrogen source, and exogenous proteins are induced to express through an inducer; finally, efficient biological synthesis of the aromadendrin is realized. The production method disclosed by the invention can realize high-efficiency production of the natural aromadendrin; within a fermentation period, namely 55h, the concentration of the aromadendrin in a fermentation solution reaches 3g / L to 5g / L and the substrate mass conversion rate reaches 70 percent to 90 percent.

Description

technical field [0001] The invention relates to a method for Escherichia coli to biosynthesize natural orangenin by utilizing naringenin, and belongs to the field of fermentation engineering. Background technique [0002] Aromadendrine, also known as Dihydrokaempferol, is a flavonol, which belongs to flavonoids and is an important precursor of plant anthocyanins. Cougarin was discovered from peach for the first time, and has since been found to exist in different plants, such as plants of the genus Tassel, Dionysus chrysanthemum, goat tree, arborvitae leaves and Siberian red pine, etc., but it is distributed in nature Not widely. [0003] Flavonoids have been widely reported to have various pharmacological activities such as anti-inflammatory, anti-oxidation, anti-tumor, and neuroprotective activities. There have been many reports showing that tangerin has various biological activities, such as anti-inflammatory activity, free radical scavenging activity and anti-tumor act...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/06C12R1/19
CPCC12P17/06C12N1/205C12R2001/19
Inventor 杨皓茹方林周睿余晓丹
Owner WUXI NEWWAY FERMENTATION TECH RES INST
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