A kind of Bacillus subtilis and the method for using Bacillus subtilis to prepare milk-flavored Cordyceps sinensis fermentation liquid

A technology of Bacillus subtilis and Cordyceps sinensis, applied in the field of microorganisms, to achieve the effects of simple production process, low production cost and easy availability of raw materials

Active Publication Date: 2022-03-08
INNER MONGOLIA KUNMING CIGARETTE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still no research on adding vanillin to Cordyceps sinensis to increase its aroma. The present invention provides a new idea for improving the aroma of Cordyceps sinensis

Method used

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  • A kind of Bacillus subtilis and the method for using Bacillus subtilis to prepare milk-flavored Cordyceps sinensis fermentation liquid
  • A kind of Bacillus subtilis and the method for using Bacillus subtilis to prepare milk-flavored Cordyceps sinensis fermentation liquid
  • A kind of Bacillus subtilis and the method for using Bacillus subtilis to prepare milk-flavored Cordyceps sinensis fermentation liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Screening and isolation of embodiment 1 bacterial strain

[0050] Sampling: In a sterile operating bench, weigh 20g of Maotai-flavored Daqu by quartering method. After grinding the sample with a mortar, wash it with 180mL of normal saline, shake the incubator at 150rpm and mix for 30min.

[0051] Bacteria enrichment culture: After a water bath, 5 mL of koji solution was drawn and inoculated into the enrichment medium, and the enrichment culture was carried out in a shaker at 37°C and 150 rpm for 24 hours.

[0052] Dilution and coating: after enrichment culture, pipette 1mL of bacterial suspension into a 9mL saline test tube to dilute. Pipette 100 μL to dilute to 10 -4 、10 -5 、10 -6 The bacterial suspension was spread on the plate isolation medium.

[0053] Isolation: Cultivate in a 37°C incubator for 24 hours, observe the colony shape after the cultivation, pick a single colony and inoculate it on the plate medium by streaking method to separate and purify.

[005...

Embodiment 2

[0055] Example 2 Fermentation method for the production of vanillin by bacterial strain biotransformation of glucose

[0056] S1: Inoculate the preserved bacterial strain on the activation medium, and culture it statically at 35-40° C. for 12-24 hours.

[0057] S2: After the seed medium is inoculated with the strain, it is cultured for 12-24 hours at a temperature of 35-40° C. and a rotation speed of 150 r / min to obtain a seed solution.

[0058] S3: After the glucose fermentation medium is inoculated with the seed solution, ferment for 100-150 hours under the culture conditions of a temperature of 35-40° C. and a rotational speed of 100-180 rpm.

[0059] Table 1 shows the activation medium and glucose fermentation medium required for the bioconversion of glucose to vanillin by the strain.

Embodiment 3

[0060] Example 3 strain producing vanillin ability test

[0061] In this experiment, the strains were preliminarily screened, and 19 strains with strong vanillin-producing ability were obtained. The specific results are shown in Table 2.

[0062] Table 2 Screening of higher yield strains

[0063]

[0064]

[0065] Take the vanillin content (x, mg / L) as the abscissa and OD280 (y) as the ordinate to draw the vanillin standard curve, see the results figure 1 . Depend on figure 1 It can be seen that the regression equation of the standard curve is y=71.716x+13.049, and the correlation coefficient R 2 = 0.9999. The vanillin content and the absorbance value showed a good linear relationship.

[0066] In this experiment, the HPLC detection of the vanillin standard was carried out with reference to the high-performance liquid chromatography method used by Zhao Jianfen et al. (05):332-333.). The resulting liquid chromatogram was figure 2 Shown, the retention time of vani...

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Abstract

The invention provides Bacillus subtilis, the strain is named Bacillus subtilis Imau 4-9, which is preserved in China Center for Type Culture Collection, and the preservation number is CCTCC NO: M2021776. The invention discloses a Bacillus subtilis (Bacillus subtilis) Imau 4-9 capable of preparing a milk-flavored Cordyceps fermented liquid, which fills in the technical gap in the preparation of the milk-flavored Cordyceps fermented liquid. The bacterial strain of the present invention can produce vanillin in glucose medium or Cordyceps sinensis medium.

Description

technical field [0001] The present application relates to the technical field of microbes, in particular to a Bacillus subtilis and a method for preparing milk-flavored Cordyceps sinensis fermentation liquid by using the Bacillus subtilis. Background technique [0002] Vanillin, chemically known as 3-methoxy-4-hydroxybenzaldehyde, found in Peruvian balsam, clove bud oil, vanilla, coffee, grapes, and brandy It has been found that it has the smell of vanilla and is sweet. As a flavoring agent, flavoring agent, coordinating agent, and flavoring agent, it is used in food, tobacco, daily chemicals and agriculture. Because of its wide range of uses, the annual demand is growing at a rate of 10%. [0003] At present, the commonly used production methods of vanillin include plant raw material extraction, chemical synthesis and biosynthesis. The content of vanillin in pod bean is about 15-20g / kg, and the planting of vanilla is limited by the region and climate environment, the yie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P1/04C12P7/24C12R1/125
CPCC12N1/20C12P1/04C12P7/24
Inventor 柴颖李力群乔月梅郭春生郝捷张善林纪旭东田数陈晨费玥高磊
Owner INNER MONGOLIA KUNMING CIGARETTE CO LTD
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