Process for synthesizing propyl gallate in organic phase by microbiological method

A technology of propyl gallate and microbial method, which is applied in the biological field, can solve problems such as difficult scale-up and industrial production, and unstable physical and chemical properties of reverse micelles, so as to save the separation and purification process, high tolerance, and reduce production cost effect

Inactive Publication Date: 2006-10-04
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 1985, Weetal successfully catalyzed the synthesis of propyl gallate and pentyl gallate for the first time with immobilized tannase, and the esterification rates were 41.4% and 78% respectively in 168 hours. In the same year, Weetal obtained the patent for the synthesis of propyl gallate. In 1989 Gaathon et al. used reverse micelles to immobilize tannase to synthesize propyl gallate, and the yield was 51% in 90 hours. However, due to the unstable physical and chemical properties of reverse micelles, it is difficult to scale up and industrialize production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Aspergillus niger (Aspergillus niger) of CGMCC preservation number No.3.315 is inoculated on the preservation medium of Aspergillus niger strain, cultivated at 30° C. for 5 days, and then the germinated Aspergillus niger spores are inoculated in the fermentation medium of 300L, wherein Spore concentration is 10 5 cells / ml, cultured for 72 hours at a stirring speed of 180 rpm and a culture temperature of 30°C.

[0018] Aspergillus niger species preservation medium is (unit: % (w / v)): 0.5% tannic acid, 0.1% peptone, K 2 HPO 4 0.1%, NaH 2 PO 4 0.1%, MgSO 4 ·7H 2 O 0.05%, agar 6%.

[0019] Aspergillus niger fermentation medium is (unit: % (w / v)): tannic acid 1.0%, soluble starch 0.2%, sucrose 0.8%, glucose 0.3%, bean cake powder 0.8%, NH 4 NO 3 0.1%, K 2 HPO 4 0.1%, KCl 0.05%, MgSO 4 ·7H 2 O 0.05%, CaCl 2 0.02%, MnCl 2 ·6H 2 O 0.04%, FeSO 4 0.001%, pH 6.0.

[0020] The above-mentioned cultured mycelia for 72 hours were collected and filtered, rinsed wi...

Embodiment 2

[0022] Aspergillus niger (Aspergillus niger) of CGMCC preservation number No.3.315 is inoculated on the preservation medium of Aspergillus niger strain, cultivated at 30° C. for 5 days, and then the germinated Aspergillus niger spores are inoculated in the fermentation medium of 300L, wherein Spore concentration is 10 5 cells / ml, cultured for 72 hours at a stirring speed of 180 rpm and a culture temperature of 30°C.

[0023] Aspergillus niger species preservation medium is (unit: % (w / v)): 0.5% tannic acid, 0.1% peptone, K 2 HPO 4 0.1%, NaH 2 PO 40.1%, MgSO 4 ·7H 2 O 0.05%, agar 6%.

[0024] Aspergillus niger fermentation medium is (unit: % (w / v)): tannic acid 4.0%, soluble starch 0.4%, sucrose 0.3%, glucose 0.1%, bean cake powder 0.8%, NH 4 NO 3 0.1%, K 2 HPO 4 0.1%, KCl 0.05%, MgSO 4 ·7H 2 O 0.05%, CaCl 2 0.02%, MnCl 2 ·6H 2 O 0.04%, FeSO 4 0.001%, pH 6.0.

[0025] The above-mentioned cultured mycelia for 72 hours were collected and filtered, rinsed with...

Embodiment 3

[0027] Aspergillus niger (Aspergillus niger) of CGMCC preservation number No.3.315 is inoculated on the preservation medium of Aspergillus niger strain, cultivated at 30° C. for 5 days, and then the germinated Aspergillus niger spores are inoculated in the fermentation medium of 300L, wherein Spore concentration is 10 5 cells / ml, cultured for 72 hours at a stirring speed of 180 rpm and a culture temperature of 30°C.

[0028] Aspergillus niger species preservation medium is (unit: % (w / v)): 0.5% tannic acid, 0.1% peptone, K 2 HPO 4 0.1%, NaH 2 PO 4 0.1%, MgSO 4 ·7H 2 O 0.05%, agar 6%.

[0029] Aspergillus niger fermentation medium is (unit: % (w / v)): 2.0% tannic acid, 0.3% soluble starch, 0.5% sucrose, 0.2% glucose, 0.9% bean cake powder, NH 4 NO 3 0.1%, K 2 HPO 4 0.1%, KCl 0.05%, MgSO 4 ·7H 2 O 0.05%, CaCl 2 0.02%, MnCl 2 ·6H 2 O 0.04%, FeSO 4 0.001%, pH 6.0.

[0030] The above-mentioned cultured mycelium for 72 hours was collected and filtered, rinsed wi...

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Abstract

The invention discloses a method for synthesizing gallic acid propyl ester in organic phase by microbe method. The method comprises the following steps: 1) collecting and filtering the mycelia cultured from aspergillus niger spores; 2) flushing with 0.85% (w / v) NaCl solution, balancing in 0.01-0.02mol / L pH 2.2-5.8 buffer solution and filtering to obtain the mycelia containing 60-80% (w / w) water; 2) in organic solvent benzene system, charging 0.06-0.1L normal propyl alcohol and 4-10mmol gallic acid into each liter organic solvent, then charging 20-50g said aspergillus niger mycelia into each liter organic solvent, implementing the biological catalytic reaction with the 150-220rpm stirring speed and under the temperature of 20-50 DEG C for 48-96 hours. The invention establishes the fermentation culture medium for the tannases of aspergillus niger, omits the separation and purification of the enzyme and lower the production cost by using the aspergillus niger as the biological catalyst instead of traditional free enzymes or fixed enzymes.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for synthesizing propyl gallate in an organic phase by a microbial method. Background technique [0002] Biotransformation belongs to the intersection field of chemistry and biology, and biocatalyst and reaction medium are the two main elements of biotransformation. Biocatalyst engineering uses fermentation technology or isolating and extracting a large number of enzymes from animals and plants, and its research has gone through the process of free enzymes, immobilized enzymes and cells; media engineering provides an ideal solvent system for the process of biocatalysts, traditionally based on Water is the solvent system. Since Klibanov et al. created the organic phase enzyme-catalyzed reaction, the organic solvent medium system has been extensively studied. [0003] The full name of tanninase is tannin acyl hydrolase (tannin acyl hydrolase, EC 3.1.1.20), which is a cell mem...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/62C12R1/685
Inventor 李永泉喻晓蔚
Owner ZHEJIANG UNIV
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