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Fungus and method for preparing vanillin from isoeugenol converted by microorgan

A technology of microbial strains and isoeugenol, which is applied in the direction of bacteria, can solve the problems of low product concentration and conversion rate, and achieve the effects of high substrate total molar conversion rate, high purity, and high product yield

Inactive Publication Date: 2005-12-28
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Available precursors include ferulic acid, eugenol, isoeugenol, curcumin, etc., and the product concentration and conversion rate are usually very low

Method used

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  • Fungus and method for preparing vanillin from isoeugenol converted by microorgan

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The soil samples contaminated by different spices were collected from a spices Co., Ltd. in Wuxi. Weighed about 10 g of each soil sample, suspended in 0.9% NaCl solution, filtered with 8 layers of gauze to remove larger impurity particles, and inoculated To enrich medium (0.2% isoeugenol; 0.5% glucose; 0.5% yeast extract; 0.5% peptone; K 2 HPO 4 ·3H 2 O 1.4%; KH 2 PO 4 0.5%; MgSO 4 ·7H 2 O 0.2%; pH 7.0), after culturing for 24 hours, continuously transfer the selection medium (without glucose, other enrichment medium) for 4 times, and spread the selective plate (the components are the same as the selection medium, containing 2% agar ). After preliminary screening, a total of 14 strains of bacteria were isolated and could grow on a plate with isoeugenol as the sole carbon source. They were all Bacillus by microscopic examination. They were transferred to the reaction solution for transformation for 3 days, and vanilla was identified by TLC. Aldehydes, found that ...

Embodiment 2

[0053] The physiological and biochemical characteristics of the screened SW-B9 strain were identified according to the eighth edition of Bergey's manual, and the results are shown in Table 3. The 16S rDNA identification was carried out according to the method of the refined molecular biology experiment guide, and the results are shown in Table 4. According to the experimental results, it can be confirmed that it belongs to Bacillus fusiformis, and it is proposed to be named Bacillus fusiformis SW-B9 (CGMCC1347).

[0054] Physiological and biochemical characteristics

[0055] +, more than 90% positive; -, more than 90% negative; D, 11%-89% positive.

[0056] 1 GACGAACGCT GGCGGCGTGC CTAATACATG CAAGTCGAGC GAACAGAGAA GGAGCTTGCT

[0057] Among them, the composition is A 25%; C 23%; G 31%; T 21%.

Embodiment 3

[0059] Carry out cultivation and fermentation with Bacillus fusiformis CGMCC1347 (SW-B9) bacterial strain, the slant medium is conventional LB medium, and seed medium is composed of: glucose 0.3%; corn steep liquor 5.5%; urea 0.3%; K 2 HPO 4 ·3H 2 O 0.09%; KH 2 PO 4 0.03%, MgSO 4 ·7H 2 O 0.1%; pH 7.0; use 250mL Erlenmeyer flask to fill 25mL culture medium, and culture at 37°C and 180r / min shaker for 12 hours as seed culture solution. The composition of the fermentation medium is: corn steep liquor 5.5%; K 2 HPO 4 ·3H 2 O 0.2%; Urea 0.1%; MgSO 4 ·7H 2 O0.1%; pH 7.5; put 50mL medium in a 250mL Erlenmeyer flask, culture at 37°C for 12 hours on a shaker at 180r / min, then add 1mmol / L vanillin as an inducer to continue culturing for 8 hours, and finally obtain stem cells containing Fermentation broth weighing 6.9 g / L.

[0060]Centrifuge the fermentation broth at 3000r / min for 10min to obtain 0.36g of wet bacteria, put 0.4mL of isoeugenol in a 250mL Erlenmeyer flask, add ...

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Abstract

Vanillin bacteria from microbial conversion isobutyl aromatic phenol and its production are disclosed. The process is carried out by fermentation culturing under optimizing condition, converting isobutyl aromatic phenol for 24í½96hrs from fermentation liquor or free cell or fixed cell with conversion liquor containing vanillic acid 2í½4g / L, conversion reacting for 72hrs in water-organic solvent double-phase system with vanillin concentration 32.5g / L in organic phase, adsorbing vanillin in conversion liquor from resin, extracting from ethyl acetate and obtaining light yellow crystalline vanillin powder. Extraction rate reaches 87%, purity of product reaches 98.1% and recovery rate reaches 93.4%.

Description

technical field [0001] The invention relates to a strain of Bacillus fusiformis CGMCC1347 (SW-B9) screened and obtained from soil, its culture and fermentation, and a method for transforming isoeugenol to prepare vanillin. Background technique [0002] Vanillin is one of the most widely used spices in the industry. It is widely used in the food industry. It can be used as the main raw material for aroma modification and fragrance fixation in food, toothpaste, soap, and tobacco; it can be used as an important raw material or intermediate in pharmaceutical and chemical industries. It can be used in the manufacture of commonly used drugs for treating hypertension, heart disease, skin diseases, and eliminating bad breath and diuresis; it can be used as a chemical auxiliary in the chemical industry, and can be used as an anti-hardening agent for plastic products and for metals such as Ni, Cr, and Cd. Electroplating brightener; in agricultural production, vanillin can be used as a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20
Inventor 孙志浩郑璞赵丽青
Owner JIANGNAN UNIV
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