Bacillus licheniformis 7172 and application thereof

A technology of Bacillus licheniformis and address, applied in the field of microorganisms, can solve the problems of difficult accumulation of vanillin and low yield of vanillin, and achieve the effects of environmentally friendly products, wide application prospects, and short production cycle

Inactive Publication Date: 2013-08-21
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the yield of vanillin transformed by most microorganisms is very low, or vanillin is quickl

Method used

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  • Bacillus licheniformis 7172 and application thereof
  • Bacillus licheniformis 7172 and application thereof
  • Bacillus licheniformis 7172 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Take a compost sample from the mushroom planting ground, weigh 6g, cut it into pieces, add 60mL of distilled water, mix it to make a suspension, and incubate at 30°C for 30min with shaking. Take the suspension prepared above and transfer it to the basic medium with ferulic acid concentrations of 0.1%, 0.2%, 0.3%, 0.4%, and 0.5%, and place the triangular flask in a water bath shaker at 50°C for oscillation Cultivate and observe its growth. Take 1mL of the above-mentioned primary screened ferulic acid concentration of 0.1%-0.5% of the culture solution, transfer to the newly prepared re-screening ferulic acid concentration of 0.4%, 0.5%, 0.6%, 0.7%, 0.8% In the culture medium, the concentration of ferulic acid was further increased, and the Erlenmeyer flask was placed in a water-bath shaker at 50°C for shaking culture. Take 10 μL of bacterial solution from the above-mentioned culture solution with a ferulic acid concentration of 0.1%-0.8%, properly dilute it with sterile ...

Embodiment 2

[0044] After the Bacillus licheniformis cells were cultured for 24 hours, they were inserted into LB medium with a volume percentage of 1% inoculum. After the cells were cultured for 20 hours, the cells were collected and added to phosphate buffer containing 5 g / L ferulic acid. Under the condition of 50°C, shake culture at 160 rpm, take samples, and quickly identify the degradation of ferulic acid and the generation of vanillin by TLC. The Bacillus licheniformis biotransformation of ferulic acid in this example produces vanillin. The detailed steps involved are as follows:

[0045] (1) Slant culture: Inoculate Bacillus licheniformis 7172 on LB solid medium, and culture it statically for 24 hours at 50°C and pH7;

[0046] (2) Preparation of seed culture solution: Inoculate the slant culture obtained in step (1) into the seed medium, culture at 50°C and 200 rpm for 24 hours, and transfer 1 to 3 times continuously to obtain the seed culture liquid;

[0047] (3) Fermentation cul...

Embodiment 3

[0052] The Bacillus licheniformis cells were inserted into the LB medium with 1% volume percentage inoculum after 24 hours of seed culture. After the cells were cultured for 20 hours, the cells were collected and added to phosphate buffer containing 5 g / L ferulic acid. Under the condition of 160rpm shaking culture at ℃ for two days, samples were taken, and the metabolites were detected and analyzed by GC-MS. The Bacillus licheniformis biotransformation ferulic acid of the present embodiment produces vanillin, and the detailed steps involved are as follows:

[0053] (1) Slant culture: Inoculate Bacillus licheniformis 7172 on LB solid medium, and culture it statically for 24 hours at 50°C and pH7;

[0054] (2) Preparation of seed culture solution: Inoculate the slant culture obtained in step (1) into the seed medium, culture at 50°C and 200 rpm for 24 hours, and transfer 1 to 3 times continuously to obtain the seed culture liquid;

[0055] (3) Fermentation culture of bacteria:...

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Abstract

The invention discloses bacillus licheniformis which is classified and named as Bacilluslicheniformis 7172 and is preserved in the common microorganism center of the microorganism strain preservation management committee in China with the preservation number of CGMCC NO.7172 in Beijing, China at January 18th in 2013. According to the bacillus licheniformis and application thereof, vanillin is produced by biologically converting ferulic acid by using the bacillus licheniformis under a high temperature condition; the growth of infectious microbes can be effectively controlled; and the industrial production is facilitated. Furthermore, by the method, the environment pollution is low, the production period is short, and a product is environment-friendly, safe and reliable; the long-time shortcoming caused by the production of the vanillin by a chemical synthesis method is overcome; and the bacillus licheniformis has a wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a bacillus licheniformis 7172 and an application thereof. Background technique [0002] Vanillin, scientific name 4-hydroxy-3-methoxybenzaldehyde, molecular formula C 8 h 8 o 3 , relative molecular weight 152.15. The pure product of vanillin is usually white or yellowish needle crystal, with the unique aroma of vanilla bean. Slightly soluble in water, easily soluble in ethanol, chloroform, ether, glacial acetic acid, pyridine and other organic solvents and strong alkali solutions. The chemical properties of vanillin are not very stable, it is easily affected by light, it is gradually oxidized to vanillic acid in the air, and it is easy to change color in alkaline solution. Non-toxic, harmless to humans and animals, and locally irritating to skin and mucous membranes. [0003] Vanillin is a broad-spectrum high-grade spice with the largest output and most u...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/24C12R1/10
Inventor 丁少军保华荣胡宏飞
Owner NANJING FORESTRY UNIV
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