CN1439713A - Microbe (No.3) and production thereof - Google Patents
Microbe (No.3) and production thereof Download PDFInfo
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- CN1439713A CN1439713A CN 02113391 CN02113391A CN1439713A CN 1439713 A CN1439713 A CN 1439713A CN 02113391 CN02113391 CN 02113391 CN 02113391 A CN02113391 A CN 02113391A CN 1439713 A CN1439713 A CN 1439713A
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- bean sprouts
- bacillus pumilus
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Abstract
A Bacillus pumilus No.3 (CGMCC No.0675) used for tobacco processing is prepared by separating it from tobacco leaf. It can decrease the irritation of cigarette smoke and the protein and nocitin of tobacco leaf, and improve the smell and taste of cigarette.
Description
Technical field
The present invention relates to microbial technology field, specifically a kind of microorganism (No.3) and preparation method thereof.
Background technology
At present because former cigarette that spontaneous fermentation is not good and blue or green cigarette when in mixing the industry prescription, using, to flue gas brought pungency big, inhale pungent, coarse, puckery mouthful of flavor, the tongue that stagnates, shortcoming such as fragrance is not enough or lack fragrance, and incendivity is bad.This has brought bigger damage not only for the aspects such as inner quality, product style of cigarette finished product.Moreover at present there are nearly 60% upper tobacco leaf and discarded tobacco leaf in each opium factory of the whole nation, tobacco company because rate of utilization is low, and a large amount of overstocking takies a large amount of warehouses and fund.
Summary of the invention
The objective of the invention is at there not being effective tobacco mellowing agent at present, microorganism of a kind of operability that can be used to improve tobacco product quality and upper tobacco leaf, discarded tobacco leaf and preparation method thereof is provided.
The present invention adopts the microorganism that separates the beneficial microorganism method of tobacco leaf own and obtain, and this microorganism is bacillus pumilus No.3 (Bacillus pumilus), and its preservation registration number is CGMCCNO.0675.
Microorganism of the present invention has following character after testing:
Cell is shaft-like, Gram-positive, and size is 0.5*1.5～2.0m, produces nearly middle ellipticity gemma of giving birth to, sporangiocyst does not expand.Poly-salt (PHB) particle is arranged in the cell.Bacterium colony on bouillon media is circle, expansion, neat in edge, and is faint yellow, glossy, and colony diameter is greater than 1 millimeter after 18 hours.Do not produce acetyl methyl carbinol (V-P reacting positive), VP liquid final pH is 7.5.The catalase positive.The lecithinase feminine gender, reduction nitrate.In pH5.7 and 7%NaCl, grow, seek strict aerobiosis.Produce acid from glucose, pectinose, wood sugar and N.F,USP MANNITOL.Litmus milk is reduction reaction, decomposes cruel plain.Utilize Citrate trianion.
Method for culturing microbes of the present invention is: adopt solid bean sprouts medium YBA earlier, with above-mentioned bacterium on the test tube slant substratum inoculation after, 27-29 ℃ of following activation culture, after two days, adopt fluid bean sprouts medium YB again, bacillus megaterium in the test tube is seeded in the juice broth of Erlenmeyer flask YB bean sprouts 28 ℃ of herbal medicine shaking tables cultivated 24 hours.
The liquid fermenting preparation method of this microorganism carries out according to the following steps:
1) test tube substratum: adopt sterilization solid bean sprouts juice (YBA) substratum, bacillus pumilus is seeded on the test tube substratum, 27-29 ℃ of following activation culture in inoculation back 48 hours;
2) enlarged culturing base: adopt sterilising liq YB bean sprouts medium, bacillus pumilus in the test tube substratum is seeded in the Erlenmeyer flask, place on the shaking table 28 ℃, shaking culture 24-46 hour, as seed liquor;
3) liquid fermentation medium: in the fermentation jar, adopt liquid YB bean sprouts juice nutrient solution, 121 ℃ of autoclavings are about 30 minutes, when nutrient solution subject to sterilization is cooled to 27.5 ℃, with the ratio inoculation fermentation liquid of the culture in the Erlenmeyer flask in 5-7%, cultivated 48 hours, and can obtain bacillus pumilus bacterium liquid for 27-28 ℃.
The separation method of this microorganism is: offal is added among the sterilized bean sprouts juice nutrient solution nutrient solution YB, after 25 ℃ of shaking tables are cultivated 24 hours, draw nutrient solution with suction pipe, adopt the dilution coating method on sterilization solid bean sprouts medium YBA, to carry out spread plate, after treating that bacterium grows, on another sterilization solid bean sprouts medium YBA, carry out method of scoring with inoculating needle and separate, must list bacterium colony pure culture bacterium after the purified cultivation.
Microorganism of the present invention is used for the tobacco machining process, as the pungency and the assorted gas that reduce cigarette smoke, increases cigarette smoke Harmony, fragrance exquisiteness and improve pleasant impression and have good effect.After adding microorganism of the present invention with certain proportion, can effectively reduce protein, nicotine equal size in upper tobacco leaf and the discarded tobacco leaf, flue gas is more coordinated, fragrance manifests, and improves pleasant impression.Microorganism of the present invention can make in the tobacco stock's upper tobacco leaf accelerate it and can make and property, saving warehouse, minimizing substantial contribution overstock, the loss that reduces stock's tobacco leaf and go mouldy and cause because of the long storage insect pest can cooperate the different cigarette of inhaling flavor, style brand of exploitation simultaneously.
Specific embodiments
Microorganism of the present invention is to separate the bacillus pumilus No.3 that obtains from tobacco leaf, and its preservation registration number is CGMCC NO.0675.
Its cultural method is: adopt solid bean sprouts medium YBA earlier, with above-mentioned bacterium on the test tube slant substratum inoculation after, 27-29 ℃ of following activation culture, after two days, adopt fluid bean sprouts medium YB again, bacillus megaterium in the test tube is seeded in the juice broth of Erlenmeyer flask YB bean sprouts 28 ℃ of shaking tables cultivated 24 hours.
Its liquid fermenting is the preparation method carry out according to the following steps:
1) test tube substratum: adopt sterilization solid bean sprouts juice (YBA) substratum, bacillus pumilus is seeded on the test tube substratum, 27-29 ℃ of following activation culture in inoculation back 48 hours:
2) enlarged culturing base: adopt sterilising liq YB bean sprouts medium, bacillus pumilus in the test tube substratum is seeded in the Erlenmeyer flask, place on the shaking table 28 ℃, shaking culture 24-46 hour, as seed liquor;
3) liquid fermentation medium: in the fermentation jar, adopt liquid YB bean sprouts juice nutrient solution, 121 ℃ of autoclavings are about 30 minutes, when nutrient solution subject to sterilization is cooled to 27.5 ℃, with the culture in the Erlenmeyer flask in 6% ratio inoculation fermentation liquid, cultivated 48 hours, and can obtain bacillus pumilus bacterium liquid for 27-28 ℃.
The separation method of mentioned microorganism is: offal is added among the sterilized bean sprouts juice nutrient solution nutrient solution YB, after 25 ℃ of shaking tables are cultivated 24 hours, draw nutrient solution with suction pipe, adopt the dilution coating method on sterilization solid bean sprouts medium YBA, to carry out spread plate, after treating that bacterium grows, on another sterilization solid bean sprouts medium YBA, carry out method of scoring with inoculating needle and separate, must list bacterium colony pure culture bacterium after the purified cultivation.
Claims (4)
1, a kind of microorganism is characterized in that it is to separate the bacillus pumilus No.3 (Bacillus pumilus) that obtains from tobacco leaf, and its preservation registration number is CGMCC NO.0675.
2, method for culturing microbes according to claim 1, it is characterized in that adopting earlier solid bean sprouts medium YBA, with above-mentioned bacterium on the test tube slant substratum inoculation after, 27-29 ℃ of following activation culture, after two days, adopt fluid bean sprouts medium YB again, bacillus pumilus in the test tube is seeded in the juice broth of Erlenmeyer flask YB bean sprouts 28 ℃ of shaking tables cultivated 24 hours.
3, the liquid fermenting preparation method of microorganism according to claim 1 is characterized in that carrying out according to the following steps:
1) test tube substratum: adopt sterilization solid bean sprouts juice (YBA) substratum, bacillus pumilus is seeded on the test tube substratum, 27-29 ℃ of following activation culture in inoculation back 48 hours;
2) enlarged culturing base: adopt sterilising liq YB bean sprouts medium, bacillus pumilus in the test tube substratum is seeded in the Erlenmeyer flask, place on the shaking table 28 ℃, shaking culture 24-46 hour, as seed liquor;
3) liquid fermentation medium: in the fermentation jar, adopt liquid YB bean sprouts juice nutrient solution, 121 ℃ of autoclavings are about 30 minutes, when nutrient solution subject to sterilization is cooled to 27.5 ℃, with the ratio inoculation fermentation liquid of the culture in the Erlenmeyer flask in 5-7%, cultivated 48 hours, and can obtain bacillus pumilus bacterium liquid for 27-28 ℃.
4, the separation method of microorganism according to claim 1, it is characterized in that offal is added among the sterilized bean sprouts juice nutrient solution nutrient solution YB, after 25 ℃ of shaking tables are cultivated 24 hours, draw nutrient solution with suction pipe, adopt the dilution coating method on sterilization solid bean sprouts medium YBA, to carry out spread plate, after treating that bacterium grows, on another sterilization solid bean sprouts medium YBA, carry out method of scoring with inoculating needle and separate, must list bacterium colony pure culture bacterium after the purified cultivation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02113391 CN1439713A (en) | 2002-02-23 | 2002-02-23 | Microbe (No.3) and production thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN 02113391 CN1439713A (en) | 2002-02-23 | 2002-02-23 | Microbe (No.3) and production thereof |
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| CN1439713A true CN1439713A (en) | 2003-09-03 |
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| CN 02113391 Pending CN1439713A (en) | 2002-02-23 | 2002-02-23 | Microbe (No.3) and production thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100343385C (en) * | 2004-11-19 | 2007-10-17 | 上海爱普香料有限公司 | Parvobacteria of high-yield 3-hydroxy-butanoic butanone |
| CN102217794A (en) * | 2011-02-23 | 2011-10-19 | 红云红河烟草(集团)有限责任公司 | Tobacco leaf biochemical additive and preparation method and application thereof |
| CN102217780A (en) * | 2011-02-23 | 2011-10-19 | 红云红河烟草(集团)有限责任公司 | Tobacco flavor extract and preparation method and application thereof |
| CN102250813A (en) * | 2011-07-17 | 2011-11-23 | 红云红河烟草(集团)有限责任公司 | Bacillus pumilus preparation for tobacco stem treatment |
| CN102250793A (en) * | 2011-06-20 | 2011-11-23 | 中国农业科学院烟草研究所 | Bacillus pumilus with anti-mildew function |
| CN102266118A (en) * | 2011-06-03 | 2011-12-07 | 川渝中烟工业公司 | Preparation method of tobacco leaves of cigars |
| CN102816710A (en) * | 2011-12-22 | 2012-12-12 | 红塔烟草(集团)有限责任公司 | Strain capable of generating protease, method for preparing protease liquid and method for improving quality of low-grade tobaccos by using protease liquid |
| CN103667120A (en) * | 2013-11-27 | 2014-03-26 | 云南省烟草农业科学研究院 | Bacillus pumilus, method for acquiring strain and application of strain in orientated degradation of nitrosoamine specifically in tobacco |
-
2002
- 2002-02-23 CN CN 02113391 patent/CN1439713A/en active Pending
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100343385C (en) * | 2004-11-19 | 2007-10-17 | 上海爱普香料有限公司 | Parvobacteria of high-yield 3-hydroxy-butanoic butanone |
| CN102217794A (en) * | 2011-02-23 | 2011-10-19 | 红云红河烟草(集团)有限责任公司 | Tobacco leaf biochemical additive and preparation method and application thereof |
| CN102217780A (en) * | 2011-02-23 | 2011-10-19 | 红云红河烟草(集团)有限责任公司 | Tobacco flavor extract and preparation method and application thereof |
| CN102266118A (en) * | 2011-06-03 | 2011-12-07 | 川渝中烟工业公司 | Preparation method of tobacco leaves of cigars |
| CN102266118B (en) * | 2011-06-03 | 2013-04-03 | 川渝中烟工业有限责任公司 | Preparation method of tobacco leaves of cigars |
| CN102250793A (en) * | 2011-06-20 | 2011-11-23 | 中国农业科学院烟草研究所 | Bacillus pumilus with anti-mildew function |
| CN102250793B (en) * | 2011-06-20 | 2013-09-25 | 中国农业科学院烟草研究所 | Bacillus pumilus with anti-mildew function |
| CN102250813A (en) * | 2011-07-17 | 2011-11-23 | 红云红河烟草(集团)有限责任公司 | Bacillus pumilus preparation for tobacco stem treatment |
| CN102816710A (en) * | 2011-12-22 | 2012-12-12 | 红塔烟草(集团)有限责任公司 | Strain capable of generating protease, method for preparing protease liquid and method for improving quality of low-grade tobaccos by using protease liquid |
| CN102816710B (en) * | 2011-12-22 | 2014-05-07 | 红塔烟草(集团)有限责任公司 | Strain capable of generating protease, method for preparing protease liquid and method for improving quality of low-grade tobaccos by using protease liquid |
| CN103667120A (en) * | 2013-11-27 | 2014-03-26 | 云南省烟草农业科学研究院 | Bacillus pumilus, method for acquiring strain and application of strain in orientated degradation of nitrosoamine specifically in tobacco |
| CN103667120B (en) * | 2013-11-27 | 2015-07-22 | 云南省烟草农业科学研究院 | Bacillus pumilus, method for acquiring strain and application of strain in orientated degradation of nitrosoamine specifically in tobacco |
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