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Bacillus methylotrophicus VJ4-1 and method for producing natural vanillin by ferulic acid biotransformation with the same

A methanol-eating spore and biotransformation technology is applied in the field of methanol-eating Bacillus and its biotransformation of ferulic acid to produce natural vanillin, which can solve the problems of complex medium components, high production cost, difficulty in separation and extraction, etc. The effect of environmental friendliness, less environmental pollution, and simple extraction steps

Inactive Publication Date: 2012-10-24
HUBEI CHINA TOBACCO IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, in Streptomyces setonii ATCC39116 reported by Rabenhorst in 1996 and Amycolatopsis sp.HR167 reported by A.Muheim in 1998, the concentration of vanillin reached 12g / L and 11.5g / L respectively, and the composition of the medium was relatively complicated, the production cost was high, and the by-products Most of them include vanillic acid, vanillic alcohol and guaiacol, etc., which brings considerable difficulties to the separation and extraction of products

Method used

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  • Bacillus methylotrophicus VJ4-1 and method for producing natural vanillin by ferulic acid biotransformation with the same
  • Bacillus methylotrophicus VJ4-1 and method for producing natural vanillin by ferulic acid biotransformation with the same
  • Bacillus methylotrophicus VJ4-1 and method for producing natural vanillin by ferulic acid biotransformation with the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] The method for producing natural vanillin by fermentation of microbial strain Bacillus methylotrophicus CCTCC VJ4-1 M 201200 of the present invention is as follows:

[0054] 1. Microbial fermentation

[0055] (1) Activation of strains: take Bacillus methylotrophicus VJ4-1CCTCC M 2012004 and inoculate it on the prepared slant medium, in a constant temperature incubator, under the conditions of 30°C and pH 7, statically cultivate for 36 hours to obtain activation Bacteria, stored in a refrigerator at 4°C;

[0056] Incline medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L.

[0057] (2) Preparation of seed liquid: use an inoculation loop to pick a ring of VJ4-1 into a 500mL Erlenmeyer flask containing 200mL liquid seed medium, and shake it in a constant temperature shaker at 30°C with a rotation speed of 100rpm for 24h; activate once;

[0058] The composition of the seed liquid medium is: maltose 50g / L, peptone 10g / L, sodium chloride 5g / L.

[0059] (3) Fermentat...

Embodiment 2

[0065] The method for producing natural vanillin by fermentation of microbial strain Bacillus methylotrophicus CCTCC VJ4-1 M 201200 of the present invention is as follows:

[0066] 1. Microbial fermentation

[0067] (1) Activation of strains: take Bacillus methylotrophicus VJ4-1CCTCC M 2012004 and inoculate it on the prepared slant medium, in a constant temperature incubator, under the conditions of 35 ° C and pH 9, statically cultivate for 72 hours to obtain activation Bacteria, stored in a refrigerator at 4°C;

[0068] Incline medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L.

[0069] (2) Preparation of seed liquid: use an inoculation loop to pick a ring of VJ4-1 into a 500mL Erlenmeyer flask containing 200mL liquid seed medium, and culture it in a constant temperature shaker at 25°C with a rotation speed of 150rpm for 24h; activate twice;

[0070] The composition of the seed liquid medium is: maltose 50g / L, peptone 10g / L, sodium chloride 5g / L.

[0071] (3) Ferm...

Embodiment 3

[0077] The method for producing natural vanillin by fermentation of microbial strain Bacillus methylotrophicus CCTCC VJ4-1 M 201200 of the present invention is as follows:

[0078] 1. Microbial fermentation

[0079] (1) Activation of strains: take Bacillus methylotrophicus VJ4-1CCTCC M 2012004 and inoculate it on the prepared slant medium, in a constant temperature incubator, under the conditions of 25°C and pH5, statically cultivate for 72 hours to obtain activation Bacteria, stored in a refrigerator at 4°C;

[0080] Incline medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L.

[0081] (2) Preparation of seed liquid: pick a ring of VJ4-1 with an inoculation loop and put it into a 500 mL Erlenmeyer flask containing 200 mL of liquid seed medium, in a constant temperature shaker, 30 ° C, 130 rpm shaking culture for 36 h; activate 4 times;

[0082] The composition of the seed liquid medium is: maltose 50g / L, peptone 10g / L, sodium chloride 5g / L.

[0083] (3) Fermentation...

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Abstract

The present invention discloses a strain of Bacillus methylotrophicus VJ4-1, and a method for producing natural vanillin by ferulic acid biotransformation with the Bacillus methylotrophicus VJ4-1, wherein a name of the strain is Bacillus methylotrophicus VJ4-1, the strain is preserved in the China center for type culture collection (CCTCC) in Wuhan University on January 16, 2012, and a preservation number is CCTCC NO:M2012004. The method of the present invention has advantages of high yield, simple process, mild reaction conditions, less environmental pollution, high product concentration, simple extraction step, clean production, environmentally friendly products, safety, reliability, wide main raw material source, low cost, and natural property, and has an industrial production prospect.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to a method for producing natural vanillin by Bacillus methanolivus and ferulic acid biotransformation. Background technique [0002] Vanillin, also known as vanillin, vanillin, etc., is the main component of vanilla bean. Its chemical name is 3-methoxy-4-hydroxybenzaldehyde, and its relative molecular weight is 152.15. 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. [0003] Vanillin is one of the largest and most widely used spices in the world. It is known as the "king of spices" and is widely used in chocolate, ice cream, candy and other foods, as well as cigarettes, beverages and high-end cosmetics. It can be used not only as a flavor enhancer and fixative, but also as a preservative, preservative and antioxidant in food. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/22C12R1/07
Inventor 李丹魏敏罗诚浩宋旭艳陈勇王洁喻林
Owner HUBEI CHINA TOBACCO IND
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