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Extraction method and application of p-hydroxybenzoic acid

A technology of p-hydroxybenzoic acid and extraction method, which is applied in the field of p-hydroxybenzoic acid extraction, can solve the problems of limited extraction technology and activity research of hydroxybenzoic acid, and achieve the effect of strengthening algae-inhibiting activity

Active Publication Date: 2020-11-06
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to overcome the above-mentioned deficiencies of the prior art, provide a kind of extraction method and application of p-hydroxybenzoic acid, aim to solve the technical problem that the extraction technology and active research of existing p-hydroxybenzoic acid are very limited, and provide Possible Application of Hydroxybenzoic Acid in Red Tide Biological Control

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  • Extraction method and application of p-hydroxybenzoic acid
  • Extraction method and application of p-hydroxybenzoic acid
  • Extraction method and application of p-hydroxybenzoic acid

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Effect test

Embodiment 1

[0032] Arenibacter sp.6A1 strain fermentation

[0033] The low-temperature-preserved marine bacteria Arenibacter sp.6A1 was revived on the plate medium, and then the revived strain was inoculated into the fermentation medium, 180rpm, 28°C for 96 hours;

[0034] Among them, the plate medium is 2216E liquid medium (recipe: peptone 5g / L, yeast extract 1g / L, ferric phosphate 0.01g / L, sea salt 20g / L) plus 1.5% agar, sterilized under high pressure at 121°C for 20 minutes The fermentation medium is 2216E liquid medium (formulation: peptone 5g / L, yeast extract 1g / L, ferric phosphate 0.01g / L, sea salt 20g / L).

[0035] After fermenting according to the above method, the fermentation liquid of marine bacteria Arenibactersp.6A1 containing p-hydroxybenzoic acid can be obtained.

Embodiment 2

[0037] Separation of p-hydroxybenzoic acid

[0038] The fermented liquid that embodiment 1 obtains is carried out following steps:

[0039] Ethyl acetate extraction: Use 1mol / L hydrochloric acid to adjust the pH of the fermented liquid to 3-4, extract with ethyl acetate, spin the extracted extract to dryness below 50°C, and repeat the extraction process three times to obtain acetic acid 2 g of the extract of the ethyl ester layer.

[0040] Gel column chromatography: dissolve the extract of the ethyl acetate layer in 2mL methanol, carry out Sephadex LH20 (column specification: diameter=3.5cm, column length=100cm) column chromatography, utilize 400mL volume ratio to be 1:1 The mixture of chloroform and methanol was used as the elution solvent for isocratic elution, each 50mL was collected as a fraction, and a total of 8 fractions were obtained, G3-A, G3-B, G3-C, G3-D, G3-E , G3-F, G3-G and G3-H.

[0041] HPLC separation: the G3-H fraction was concentrated and evaporated to dr...

Embodiment 3

[0049] Identification of algae-dissolving activity of p-hydroxybenzoic acid

[0050] Algae-dissolving experiments were carried out by setting gradient concentrations of p-hydroxybenzoic acid. Count the algae liquid through the plankton box to measure the algal concentration, shake up and divide the algal liquid; dissolve p-hydroxybenzoic acid to a sample concentration of 100 μg / μl with DMSO, and then serially dilute to 50 μg / μl, 25 μg / μl, 12.5μg / μl and 6.75μg / μl, take 1ml / well of the algae liquid into a 24-well plate, add 1ul of the prepared sample to the final concentration of the sample to 100μg / ml, 50μg / ml, 25μg / ml, 12.5μg / ml, 6.75μg / ml, add 1μl DMSO to the control group, set up 3 repetitions for each experimental group and control group; put the 24-well plate in the light incubator for culture; use planktonic counting plate at 15min and 120min Count the number of algae. figure 1 It shows the influence of the samples of various concentrations on the cell number of Hakata...

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Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to an extracting method and an application of p-hydroxybenzoic acid. The extracting method comprises the following steps: inoculating the marine bacteria Arenibacter sp.6A1 into a fermentation culture medium, and carrying out fermentation culture to obtain a fermentation liquor; adjusting the pH value of the fermentation liquor to 3-4, and then carrying out extraction treatment to obtain an extract; and dissolving the extract, sequentially performing gel column chromatography and high performance liquid chromatography (HPLC) separation to obtain the p-hydroxybenzoic acid, wherein the HPLC separation conditions are as follows: gradient elution is carried out with a methanol / water mixed solvent, the detection wavelength is 254 nm, and the time of collecting liquid phase peaks is 25 min. Through the method, hydroxybenzoic acid with high purity and algae-dissolving activity is finally obtained, and a good foundation is laid for industrial production of hydroxyl benzoic acid.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to an extraction method and application of p-hydroxybenzoic acid. Background technique [0002] Red tide refers to the harmful phenomenon of seawater discoloration caused by the excessive proliferation of some algae, protozoa or bacteria in the ocean under certain specific climatic conditions. At present, the vast majority of red tides are caused by algae. Due to the different types of algae, in addition to red, the seawater can also appear yellow, brown or green or change color during the outbreak period. Therefore, red tides and freshwater blooms are also called harmful algal blooms ( Harmful Algal Bloom, HAB). The harm caused by red tide to marine organisms and humans can be divided into three aspects. First, red tide algae will consume oxygen in seawater, or red tide algae cells will directly block the respiratory organs of marine organisms and cause hypoxia ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07C51/42C07C65/03A01N37/40A01P13/00
CPCA01N37/40C07C51/42C07C65/03
Inventor 陈辉蓉童晶伍嘉慧胡章立黎双飞王立岩
Owner SHENZHEN UNIV
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