A method for producing 4-o-desmethylbarba lichenic acid by using Aspergillus terreus strain and then synthesizing and synthesizing oakmoss

A technology for synthesizing oak moss with methyl balba, applied in the field of microbial fermentation technology and chemical synthesis, and new strains, can solve the problems of high cost, restricting the development of synthetic oak moss, complex preparation process, etc., to improve efficiency and simplify synthesis steps and technological process, the effect of cost reduction

Active Publication Date: 2022-04-26
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in actual production, the preparation process of these two key intermediates is complicated, and the yield is low and the cost is high. Therefore, the method of producing synthetic oakmoss by chemical methods has certain defects in practical applications, which limits the synthesis Oakmoss Market Development

Method used

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  • A method for producing 4-o-desmethylbarba lichenic acid by using Aspergillus terreus strain and then synthesizing and synthesizing oakmoss
  • A method for producing 4-o-desmethylbarba lichenic acid by using Aspergillus terreus strain and then synthesizing and synthesizing oakmoss
  • A method for producing 4-o-desmethylbarba lichenic acid by using Aspergillus terreus strain and then synthesizing and synthesizing oakmoss

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Screening of an Aspergillus terreus strain capable of producing 4-O-desmethylbarbalicic acid.

[0072] The needlefish (Alennes anastomella) frozen at -20°C from the waters of Yangma Island in Muping, Yantai was sterilized with 75% ethanol on the surface, and part of the internal organs of the fish were taken under aseptic conditions for grinding. Then use sterile water to make different gradient dilutions (V / V% 0, 10, 100, 1000), and apply the stock solution and dilutions of different concentrations to ampicillin (0.05g / L) and streptomycin (0.03 g / L) on the PDA plate of two kinds of antibiotics, culture at 28 ℃, observe whether there is the growth of fungal colony every other day in the process of cultivating, and pick single colony, carry out purification.

[0073] The different fungi obtained by separation and purification were cultured on PDA plates for 7 days. After the spores grew, the spore concentration was calculated by the hemocytometer plate counting method, a...

Embodiment 2

[0080] Example 1 The application of Aspergillus terreus MEFC06, which produces 4-O-desmethylbarbalicic acid obtained through screening, in the fermentative production of 4-O-desmethylbarbalicic acid.

[0081] Include the following steps:

[0082] Step 1, preparation of Aspergillus terreus seed liquid, inoculating the seed liquid into the fermentation medium for fermentation and cultivation;

[0083] Step 2, separating and purifying the fermentation broth to obtain 4-O-desmethylbarbalicic acid.

[0084] The preparation of the seed solution described in step 1 is specifically: inoculate Aspergillus terreus MEFC06 on the PDA plate medium, cultivate at 28°C for 4-9 days, grow spores, inoculate the spores on the plate into the seed medium, inoculate Quantity is 10 6 -10 8 Each / 35mL, 28°C, 220rpm shaker culture for 2 days, to obtain seed liquid.

[0085] In step 1, the inoculation amount of the seed solution inoculated into the fermentation medium is 3:35 according to the volume...

Embodiment 3

[0089] Example 3 The method for preparing synthetic oakmoss by chemical synthesis using 4-O-demethylbarbalicic acid as raw material

[0090] Step (1) Obtain 2,4-dihydroxy-3,6-dimethylbenzoic acid by acid hydrolysis or alkali hydrolysis of 4-O-desmethylbarbaric acid;

[0091] Step (2) 2,4-dihydroxy-3,6-dimethylbenzoic acid is methylated to obtain methyl 2,4-dihydroxy-3,6-dimethylbenzoate.

[0092] This embodiment is specifically:

[0093]Add 2.2g (6.3mmol) solid crystals of 4-O-demethylbarbalicic acid in the there-necked flask, then add 10ml of catalytic amount (dissolving the solid crystals completely) of concentrated sulfuric acid with a mass fraction of 98%, in a water bath at 26°C Under stirring reaction, and adopt the HPLC analysis method reaction monitoring of 2,4-dihydroxyl-3,6-dimethylbenzoic acid and 2,4-dihydroxyl-3,6-dimethylbenzoate, the result shows The reaction was complete after 10 minutes, and no by-products were produced until 40 minutes after the late reacti...

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Abstract

The invention discloses a method for producing 4‑O‑demethylbarbalicic acid by using Aspergillus terreus strains to synthesize oakmoss, and relates to the technical fields of new strains, microbial fermentation technology and chemical synthesis. The Aspergillus terreus strains are soil Aspergillus terreus (Aspergillus terreus) MEFC06, fermentation culture of Aspergillus terreus species; separation and purification to obtain 4‑O‑desmethyl barbalicic acid, hydrolysis of 4‑O‑desmethyl barbalic acid to 2,4‑dihydroxy‑ 3,6-dimethylbenzoic acid; using a methylating agent to carry out methylation reaction, after the reaction, treatment can obtain 2,4-dihydroxy-3,6-dimethylbenzoic acid methyl ester (synthetic oakmoss ). A new method for producing synthetic oak moss by combining microbial fermentation and chemical synthesis is provided, which is green and ecological, has short synthetic route, simple process and high yield.

Description

technical field [0001] The invention relates to the technical fields of new bacterial strains, microbial fermentation technology and chemical synthesis, in particular to a method for producing 4-O-desmethylbarbalicic acid and then synthesizing oakmoss by using Aspergillus terreus strains. Background technique [0002] Oakmoss is a rare lichen resource, which generally refers to Quercus chinensis of the Usneaceae genus that grows on the trunk and branches of oak trees. Oakmoss extract and essential oil are widely used in perfumes, skin care products, cigarettes and Incense and other daily necessities, at the same time, oakmoss is also used as a food additive in beverages, baked goods, puddings, fudge, etc. Natural oakmoss is mainly produced in central and southern Europe and North Africa, a generation of Mediterranean countries, such as France and Italy. Oakmoss resources in my country are extremely scarce, and only Oakmoss I and II have been developed in Yunnan. Due to the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/62C12P7/40C12R1/66
CPCC12P7/40C12P7/62
Inventor 吕雪峰黄雪年张伟李鸿成
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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