Amycolatopsis sp. and application thereof

A technology of pseudomycobacteria and strains, applied in the field of microorganisms, can solve the problems of many by-products, difficulty in separating and purifying vanillin, and increasing production costs.

Active Publication Date: 2021-11-12
SHAANXI HEALTHFUL BIOLOGICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The molar conversion rates of the vanillin-producing strains disclosed above are all lower than 80%, and there are many by-products, including vanillic acid, vanillyl alcohol, guaiacol, etc., which bring certain challenges to the subsequent separation and purification of vanillin. Difficulty, and cause the resource waste of substrate ferulic acid, improved production cost

Method used

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  • Amycolatopsis sp. and application thereof
  • Amycolatopsis sp. and application thereof
  • Amycolatopsis sp. and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Solid medium: glucose 4g / L, yeast extract powder 4g / L, malt extract 10g / L, agar 20g / L, and the rest is water. Example 1 Screening of Amycolatopsis sp. HM-141

[0033] Soil samples collected from around fruit trees in Xi'an, Xianyang, Weinan and other places. The 10 soil samples collected were diluted to an appropriate multiple, spread on the enrichment medium, and placed in an incubator at 30°C for cultivation. After culturing for 96 hours, pick a solution capable of making 2,4-dinitrophenylhydrazine (1.6mg 2,4-dinitrophenylhydrazine / 1mL 50%H 2 SO 4 ) red colony, inoculated into the fermentation medium at 30°C, cultured at 200rpm for 24h, then added 10g / L ferulic acid, continued shaking culture for 48h, detected the fermentation liquid by HPLC, and selected the strain producing more vanillin , to obtain the original strain of transforming ferulic acid to produce vanillin.

[0034] The original strain is a Gram-positive bacterium, the thallus is filamentous, aerobic,...

Embodiment 2

[0044] Example 2 Determination of vanillin produced by Amycolatopsis sp.HM-141

[0045] Take a tube of glycerol strain Amycolatopsis sp.HM-141 preserved at -80°C, dilute and spread on a solid plate, place the plate upside down in a 30°C incubator and cultivate for 3-4d until colonies grow.

[0046] Pick a loop of bacteria from the activated plate into the seed medium, and culture at 30°C for 48-72h with a rotation speed of 200rpm.

[0047] The seed liquid was inoculated into a 5L fermenter (containing 4L fermentation medium) according to the inoculum amount of 5%, and fermented and cultivated at 30° C., the stirring speed was 800 rpm, and the aeration ratio was 1 vvm. After cultivating for 24h, add substrate ferulic acid 22.5g / L (total 90g, calculate the ferulic acid concentration according to the fermented liquid volume of 4L, ferulic acid is dissolved in 0.5M NaOH solution and its concentration is 100g / L), Then adjust the pH to 8.2, and continue to ferment for 48h.

[0048...

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Abstract

The invention relates to amycolatopsis sp. and application thereof. The amycolatopsis sp. HM-141 (the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.22871) disclosed by the invention is applied to production of vanillin by taking ferulic acid as a substrate. Experiments confirm that the amycolatopsis sp. HM-141 uses ferulic acid as a substrate to produce vanillin, the molar conversion rate can reach up to 87%, impurities (or byproducts) in the product are obviously lower than that in the prior art, the detection amount of vanillin alcohol is 0, the detection amount of vanillic acid is as low as 0.25 g/L, and the amycolatopsis sp. HM-141 has obvious advantages.

Description

【Technical field】 [0001] The invention relates to the technical field of microorganisms, in particular to a strain of Amycolatopsis sp., and also relates to a method for biotransformation of the Amycolatopsis sp. to produce vanillin by using ferulic acid as a substrate. 【Background technique】 [0002] Vanillin, also known as vanillin and vanillin, is one of the most widely used seasonings in the world and is widely used in food, medicine, cosmetics, agriculture and other fields. Vanillin has the reputation of "the king of food spices". It has vanilla bean aroma and rich milky aroma, and can play a role in enhancing fragrance and flavor in food; in medicine, vanillin is a synthetic compound An important raw material for medicines; vanillin can be used as a flavoring agent in cosmetics, perfumes and other fields; it can also be used as a ripening agent and yield-increasing agent for crops in agricultural production. Due to its wide range of applications, the vanillin produced...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/24C12R1/01
CPCC12N1/20C12P7/24
Inventor 孟永宏强珊郭建琦牛永洁杨璐
Owner SHAANXI HEALTHFUL BIOLOGICAL ENG
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