Bacillus pumilus and application of the same in producing natural vanillin by biologically converting iso-eugenol

A Bacillus pumilus, biotransformation technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, applications, etc., can solve the problems of low conversion rate, low vanillin concentration, high cost of vanillin, etc., and achieve by-product Fewer, simple extraction steps, low price effect

Active Publication Date: 2007-11-28
SHANGHAI KAIXIN BIOTECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In view of the fact that the demand for natural vanillin is increasing year by year, the cost of producing vanillin with natural ferulic acid as a substrate is too high, and the production process with eugenol as a substrate has low vanillin concentration and low conversion rate. The present invention provides A strain of Bacillus pumilus pumilus S-1 CCTCC M 205165 screened from soil, and a method for producing high-concentration natural vanillin with low-cost natural isoeugenol as a substrate with high conversion rate using the strain

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  • Bacillus pumilus and application of the same in producing natural vanillin by biologically converting iso-eugenol

Examples

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Embodiment 1

[0024] Example 1: Screening of Bacillus pumilus S-1 CCTCC M 205165

[0025]Take the soil around the clove tree, weigh 5 grams and dissolve it in 100 milliliters of normal saline, mix well, take 10 milliliters and add it to 50 milliliters of the transformation medium, at 30 °C, shake at 120 rpm for 24 hours, then add mass Isoeugenol with a volume ratio of 1% was shaken at 120 rpm for 48 hours, and this process was repeated 3 to 5 times. The cultured bacterial solution was diluted 1,000,000 times and spread on LB solid medium added with 1% glucose by mass volume ratio, and cultured statically at 30° C. for 20 hours. Select the grown single colony and inoculate it into 50 ml of transformation medium, add 1% isoeugenol in a mass-volume ratio after shaking for 24 hours, and shake at 120 rpm for 120 hours to screen for the ability to transform isoeugenol into vanillin Finally, a strain with better degradation ability and a large amount of vanillin accumulation was obtained, that is...

Embodiment 2

[0030] Bacillus pumilus S-1 CCTCC M 205165 cells were inserted into the transformation medium with an inoculum of 6% by volume after 15 hours of seed culture, and 10 g / L isoeugenol was added after the cells were cultured for 25 hours. Shake culture at 120 rpm. After 120 hours, samples were taken to determine the concentration of vanillin.

[0031] The Bacillus pumilus S-1 CCTCC M 205165 involved in this embodiment converts isoeugenol to produce natural vanillin, and the detailed steps involved are as follows:

[0032] (1) Slant culture: Inoculate Bacillus pumilus S-1 CCTCC M 205165 on LB solid medium supplemented with 1% glucose, and culture at 30°C for 20 hours;

[0033] (2) Preparation of seed culture solution: use the slant culture obtained in step (1) to inoculate into LB liquid medium with 1% glucose in mass volume ratio, and culture at 30°C with shaking at 120 rpm for 15 hours , to prepare seed culture solution;

[0034] (3) Transformed cell culture: use the seed cult...

Embodiment 3

[0040] Bacillus pumilus S-1 CCTCC M 205165 cells were inserted into the transformation medium with an inoculum of 6% by volume after 15 hours of seed culture, and 10 g / L isoeugenol was added after the cells were cultured for 35 hours. Shake culture at 120 rpm. After 120 hours, samples were taken to determine the concentration of vanillin.

[0041] The Bacillus pumilus S-1 CCTCC M 205165 involved in this embodiment converts isoeugenol to produce natural vanillin, and the detailed steps involved are as follows:

[0042] (1) Slant culture: Inoculate Bacillus pumilus S-1 CCTCC M 205165 on LB solid medium supplemented with 1% glucose, and culture at 30°C for 20 hours;

[0043] (2) Preparation of seed culture solution: use the slant culture obtained in step (1) to inoculate into LB liquid medium with 1% glucose in mass volume ratio, and culture at 30°C with shaking at 120 rpm for 15 hours , to prepare seed culture solution;

[0044] (3) Transformed cell culture: use the seed cult...

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Abstract

The invention discloses Bacillus pumilis and its application of biotransformation of iso-eugenol into natural vanillin. The name of bacterial stripe is Bacillus pumilus S-1, preserved in China Center for Type Culture Collection on 3rd, January, 2006 and its serial number is CCTCC M 205165. The procedure of transforming iso-eugenol into natural vanillin includes: (1)cultivation, (2)preparation of seed culture fluid, (3)culture of transforming cell, (4) transformation.The invention exists in field of biology and technology. It can improve the transformating rate of vanillin with high concentration, solving problems of low concentration and low yield of vanillin by biological method.

Description

technical field [0001] The invention relates to a method for biotransforming isoeugenol to produce natural vanillin, in particular to a method for using a strain of bacillus pumilus to biotransform isoeugenol to produce natural vanillin. Background technique [0002] Vanillin, the chemical name is 4-hydroxy-3-methoxybenzaldehyde (4-hydroxy-3-methoxybenzaldehyde), also known as vanillin, vanillin. The English name is vanillin, and the molecular formula is C 8 h 8 0 3 , the molecular weight is 152.14. The appearance of vanillin is white to light yellow needle-like crystal or powder. It has the unique aroma of vanilla bean and is slightly sweet. It is slowly oxidized to vanillic acid in humid air. The boiling point is 284-285°C, the relative density is 1.056, slightly soluble in water, easily soluble in organic solvents such as ethanol, ether, chloroform, glacial acetic acid, carbon disulfide and pyridine. Vanillin has low toxicity, rat oral LD 50 1580 mg / kg. [0003] Va...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/31C12P1/04C12P7/24C12R1/08
Inventor 许平华栋梁林山杜毅魏中浩陈洪
Owner SHANGHAI KAIXIN BIOTECH
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