Bacillus methylotrophicus and preparation method as well as application of tobacco cimicifugae extract

A technology of methanol-eating spores and bacillus, which is applied in the field of preparation of methanol-eating bacillus and its Cimicifuga extract for tobacco, which can solve the problems of single aroma and taste, complicated extraction steps, and insufficient aroma intensity of the extract for tobacco, and achieve technological The effect of simple process, wide sources and wide sources of raw materials

Inactive Publication Date: 2012-08-01
HUBEI CHINA TOBACCO IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the defects that the extraction steps of chemically synthesized tobacco extract are complex, the environment is seriously polluted, the aroma and taste of the tobacco extract is single and the aroma intensity is still insufficient, the present invention provides a kind of Bacillus methylotrophicus, and the strain is named Bacillus methylotrophicus VJ4-1 was preserved in the Chinese Type Culture Collection Center of Wuhan University on January 11, 2012, with the preservation number CCTCC NO: M2012004. Its biological characteristics are: it is a Gram-positive bacterium that can grow in 10% NaCl, It has the activity of arginine dihydrolase and ornithine decarboxylase, which can produce acid and liquefy milk; Inositol, sucrose, maltose, D-sorbitol, L-arabinose, DL-lactate, L-alanine glycogen, L-proline histidine, citrate as carbon source medium growth, its 16S rRNA sequence is 100% similar to the 16S rRNA sequence of Bacillus methylotrophicus

Method used

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  • Bacillus methylotrophicus and preparation method as well as application of tobacco cimicifugae extract
  • Bacillus methylotrophicus and preparation method as well as application of tobacco cimicifugae extract
  • Bacillus methylotrophicus and preparation method as well as application of tobacco cimicifugae extract

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Embodiment 1

[0034] The present invention utilizes the microorganism Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 to biotransform the preparation method of chaff extract for tobacco as follows:

[0035] 1) Crush Cimicifuga to 10 meshes, reflux extraction with water at a mass ratio of 1:20, filter for 3 hours, and sterilize the filtrate at 115°C for 25 minutes;

[0036] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 on the potato dextrose agar slant medium, and culture it statically for 24 hours at 25°C and pH 10, then place it in the refrigerator;

[0037] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;

[0038] 3) Preparation of seed culture solution: use the slant culture obtained in the above step 2) to inoculate the seed culture medium, shake and culture for 12 hours at 25° C. with a rotation speed of 150 rpm, and transfer 4 times continuously to obtain the seed culture solution;

[0039] The composition of the seed liquid medium...

Embodiment 2

[0043] The present invention utilizes the preparation method of microorganism Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 biotransformation tobacco chaff extract as follows:

[0044] 1) Crush Cimicifuga to 100 mesh, reflux extraction with water at a mass ratio of 1:30, filter for 2 hours, and sterilize the filtrate at 115°C for 25 minutes;

[0045] 2) Slant culture: inoculate Bacillus methanolivorus on the potato dextrose agar slant medium, and place it in the refrigerator after standing for 36 hours at 30°C and pH7;

[0046] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;

[0047] 3) Preparation of seed culture solution: use the slant culture obtained in the above step 2) to inoculate the seed culture medium, vibrate and culture at 30° C. with a rotation speed of 120 rpm for 24 hours, and make the seed culture solution after continuous transfer for 3 times;

[0048] The seed liquid medium consists of: maltose 50g / L, peptone 10g / L, sodium chloride 5g / ...

Embodiment 3

[0052] The present invention utilizes the microorganism Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 to biotransform the preparation method of chaff extract for tobacco as follows:

[0053] 1) Crush Cimicifuga to 200 mesh, reflux extraction with water at a mass ratio of 1:40 for 1 hour, filter, and sterilize the filtrate at 115°C for 25 minutes;

[0054] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 on the potato dextrose agar slant medium, and culture it statically for 72 hours at 35°C and pH 5, then place it in the refrigerator;

[0055] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;

[0056] 3) Preparation of seed culture solution: use the slant culture obtained in the above step 2) to inoculate the seed culture medium, vibrate and culture at 35° C. with a rotation speed of 100 rpm for 36 hours, and make the seed culture solution after two consecutive transfers;

[0057] The seed liquid medium consists of: maltose 5...

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Abstract

The invention discloses bacillus methylotrophicus, wherein the name of the strain is bacillus methylotrophicus VJ4-1 with a collection number of CCTCC NO: M 2012004. The invention provides biotransformation of tobacco rhizoma cimicifugae extract by using microbial bacillus methylotrophicus and application of the tobacco rhizoma cimicifugae extract to tobacco. Natural tobacco flavor can be produced by fermenting rhizoma cimicifugae through microbial strain. The method has the advantages of simple process, wide source of raw materials, environmental friendliness and harmlessness in a productionprocess and industrial application prospect; and the obtained natural flavor extract can be directly used for flavoring of tobacco, has the advantages of coordination with smoke, light sweet flowery flavor, rich aroma, softness, comfort, good aftertaste and capability of improving smoke flavor and can be mixed with other tobacco essence and perfumes for use.

Description

technical field [0001] The invention relates to the technical field of spices for tobacco, in particular to a preparation method and application of bacillus methanolate and cohosh extract for tobacco. Background technique [0002] In tobacco products, the aroma and taste of tobacco are very important. Adding spices is a key link in the cigarette process, and it is also an effective means to improve the product style and improve the quality of cigarette aroma. Tobacco flavors can be divided into chemical synthetic flavors and natural flavors. In recent years, the research and development of natural plant flavors has been the focus and hotspot of ingredients because of their Chinese characteristics, natural harmlessness, and easy availability of raw materials. The development of plant-extracted spices has limitations due to the limited available plant resources. It is a good idea to apply microbial fermentation engineering to the biosynthesis of flavors and fragrances. Using ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A24B3/12C12R1/07
Inventor 罗诚浩魏敏宋旭艳陈义坤李冉
Owner HUBEI CHINA TOBACCO IND
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