Process of synthesizing gastrodiacin by microorganism cell biological transferring parhydroxy benzyl methylol

A technology of p-hydroxybenzyl alcohol and microbial cells, which is applied in the field of bioengineering and microbial fermentation, can solve the problems of gastrodin biosynthesis, high requirements for cell culture, long conversion time, etc., and achieve substrate conversion efficiency High, simple cell culture, and short culture time

Inactive Publication Date: 2007-07-18
ZHEJIANG UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

However, there are also great defects in the conversion and synthesis of gastrodin by plant cells. Not only the cell culture requires high conditions, the cycle is long, the required transformation time is long, and the yield is low. no substantial progress

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1 The cultivation of Armillaria chrysogreen ZJUQH mycelium

[0022] Firstly, the biotransformed cells are obtained by culturing seeds and fermentation media, and the key step is to obtain a large number of multiplication of fungal mycelia. The specific cultivation method is as follows:

[0023] Slope seed medium: adopt glucose yeast extract solid medium (glucose 15g / L, yeast extract 2g / L, dipotassium hydrogen phosphate 0.5g / L, potassium dihydrogen phosphate 0.1g / L, magnesium sulfate 0.3g / L, 2 % agar, natural pH), cultivated at 18-20°C for 3-5 days before use.

[0024] Liquid seed medium: adopt glucose yeast extract basal liquid medium (glucose 15g / L, yeast extract 2g / L, dipotassium hydrogen phosphate 0.5g / L, potassium dihydrogen phosphate 0.1g / L, magnesium sulfate 0.3g / L, pH naturally).

[0025] Seed shake flask fermentation culture method: the fermentation temperature is 23°C, the rotation speed is 120r / min, the fermentation is stopped after 3 days of cul...

Embodiment 2

[0030] Example 2 The experiment of biosynthesizing gastrodin by Armillaria chrysanthemum ZJUQH mycelium cells with different culture time

[0031] According to the method in Example 1, Armillaria chrysanthemum ZJUQH cells pre-cultured for 1-6 days were respectively obtained, and used for the conversion of p-hydroxybenzyl alcohol to synthesize gastrodin.

[0032] With the sterilized potato dextrose liquid (the potato dextrose liquid contains the potato of mass percentage concentration 2% and the glucose of 0.5%) as basal medium, in every 100ml basal medium, Armillaria chrysanthemum ZJUQH wet cell addition 10g, to Hydroxybenzyl alcohol is first formulated into a 10mmol / L aqueous solution and sterilized by filtration with a 0.22 μm microporous membrane. Based on the volume of the entire conversion reaction system, 1.5 mmol of p-hydroxybenzyl alcohol is added to each liter of the conversion reaction system, and the conversion time is 2 days , the biotransformation temperature is 2...

Embodiment 3

[0036] Example 3 Synthesis of gastrodin with different concentrations of substrate (p-hydroxybenzyl alcohol)

[0037] According to the method in Example 1, Armillaria chrysanthemum ZJUQH cells pre-cultured for 3 days were obtained, and used for the conversion of p-hydroxybenzyl alcohol to synthesize gastrodin.

[0038] Using sterilized potato glucose liquid as the base medium, p-hydroxybenzyl alcohol was first prepared into a 10mmol / L aqueous solution and sterilized by filtration with a 0.22μm microporous membrane. The amount of bacteria ZJUQH wet cells added is 1-10g / 100ml of basal medium, the transformation time is 2 days, the biotransformation temperature is 23°C, and the shaker speed is 120r / min. The specific experimental results are shown in Table 2 below:

[0039] Table 2 Synthesis of gastrodin with different concentrations of substrates

[0040] Substrate concentration (mol / L conversion

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Abstract

The present invention discloses microbe cell synthesis process of converting p-hydroxy benzyl alcohol to synthesize gastrodine. The process is one converting reaction of wet cell of rmillaria luteo-virens ZJUQH in 1-10 g / 100ml and 0.22 micron microporous film filtered p-hydroxy benzyl alcohol in 0.1-3.1 mmol / L, in the culture liquid comprising sterilized potato in 2 wt% concentration and glucose in 0.5 concentration, at temperature of 20-25 deg.c and shaking table rotation speed of 50-200 rpm for 1-3 days. Compared with available plant cell conversion process, the microbe cell synthesis process has the advantages of short period, simple operation and easy control.

Description

technical field [0001] The invention relates to high-density cultivation of plateau rare edible fungi by using bioengineering technology, and using the cultured cells to biosynthesize gastrodin with p-hydroxybenzyl alcohol as a substrate, and belongs to the field of bioengineering and microbial fermentation. Background technique: [0002] Gastrodin, namely 4-hydroxymethylbenzene-β-D-glucopyranoside or p-hydroxymethylbenzene-β-D-glucopyranoside, is the main active ingredient of the Orchidaceae plant Gastrodia elata Blume , has sedative, anti-convulsant, anti-inflammatory and enhanced immune function. It is widely used clinically for vertigo (Ménière's disease, viral vertigo, vestibular neuronitis, occipital neuralgia, etc.), headache (neurasthenia and neurasthenia syndrome, vascular headache, tension headache, brain trauma syndrome) syndrome, migraine, etc.) and adjuvant treatment of epilepsy. [0003] Biotransformation is a biochemical reaction that uses plant isolated cel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/44C12R1/645
Inventor 陈启和何国庆刘彩琴朱建良
Owner ZHEJIANG UNIV
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