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Preparation method and application of Bacillus methylotrophicus and tobacco chaff extract thereof

A technology of methanol-eating spores and bacilli, which is applied in the field of preparation of methanol-eating bacillus and its chaff extract for tobacco, can solve the problems of single aroma and taste of tobacco extract, insufficient aroma intensity, complicated extraction steps, etc., and achieve increased Characteristic aroma, rich smoke aroma, and low cost of raw materials

Inactive Publication Date: 2013-11-13
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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  • Preparation method and application of Bacillus methylotrophicus and tobacco chaff extract thereof
  • Preparation method and application of Bacillus methylotrophicus and tobacco chaff extract thereof
  • Preparation method and application of Bacillus methylotrophicus and tobacco chaff extract thereof

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Embodiment 1

[0032]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:

[0033] 1) The chaff and water were refluxed and extracted at a mass ratio of 1:20 for 3 hours, then filtered, and the filtrate was sterilized at 115°C for 25 minutes;

[0034] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1 CCTCCNO: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;

[0035] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;

[0036] 3) Preparation of seed culture medium: use the slant culture obtained in the above step 2) to inoculate the seed culture medium, vibrate for 12 hours at 25°C and rotate at 150 rpm, and transfer 4 times in succession to prepare the seed culture medium;

[0037] The composition of the seed liquid medium is: maltose 50g / L, peptone...

Embodiment 2

[0041] The present invention utilizes microorganism Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 to biotransform the preparation method of chaff extract for tobacco as follows:

[0042] 1) The husk and water were refluxed and extracted at a mass ratio of 1:30 for 2 hours, then filtered, and the filtrate was sterilized at 115°C for 25 minutes;

[0043] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1CCTCCNO:M 2012004 on the potato dextrose agar slant medium, and culture it statically for 36 hours at 30°C and pH 7, then place it in the refrigerator;

[0044] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;

[0045] 3) Preparation of seed culture solution: use the slant culture obtained in the above step 2) to inoculate the seed medium, vibrate and culture at 30°C and 120 rpm for 24 hours, and make the seed culture solution after continuous transfer for 3 times;

[0046] The seed liquid medium consists of: maltose 50g / L, peptone 10g / L, sodium ch...

Embodiment 3

[0050] 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:

[0051] 1) The husk and water were refluxed and extracted at a mass ratio of 1:40 for 1 hour, then filtered, and the filtrate was sterilized at 115°C for 25 minutes;

[0052] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1CCTCCNO: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;

[0053] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;

[0054] 3) Preparation of seed culture solution: use the slant culture obtained in the above step 2) to inoculate the seed medium, vibrate and culture at 35°C and rotate at 100 rpm for 36 hours, and make the seed culture solution after two consecutive transfers;

[0055] The seed liquid medium consists of: maltose 50g / L, peptone 10g / L, so...

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Abstract

The invention discloses a Bacillus methylotrophicus strain VJ4-1 of which the preservation number is CCTCC No:M 2012004. The invention provides a tobacco chaff extract prepared by using microbe Bacillus methylotrophicus and conducting biotransformation and an application of the chaff extract in tobacco. According to the invention, by applying the fermented and purified tobacco chaff extract in cigarettes, the extract is in harmony with the smoke, reduces miscellaneous gases, reduces the irritancy, obviously adding characteristic fragrance, enriches the tobacco aroma, and improves the tasting; the method has the advantages of simple process, mild reaction conditions, low pollution, high concentration of the products, simple extraction steps, clean production, environmentally friendly product, safety and reliability, wide sources of main raw materials, extremely low cost, and nature, and has an industrial production prospect.

Description

technical field [0001] The invention relates to the technical field of flavorings for tobacco, in particular to a preparation method and application of Bacillus methanolivorus and chaff 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. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A24B3/12C12R1/07
Inventor 魏敏罗诚浩宋旭艳李冉刘华臣
Owner HUBEI CHINA TOBACCO IND
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