CN108642104A - A kind of bacteria cellulose, preparation method and applications - Google Patents
A kind of bacteria cellulose, preparation method and applications Download PDFInfo
- Publication number
- CN108642104A CN108642104A CN201810270577.2A CN201810270577A CN108642104A CN 108642104 A CN108642104 A CN 108642104A CN 201810270577 A CN201810270577 A CN 201810270577A CN 108642104 A CN108642104 A CN 108642104A
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- Prior art keywords
- bacteria cellulose
- tobacco
- prepared
- tobacco material
- culture medium
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention belongs to technology field of tobacco preparation, and in particular to a kind of bacteria cellulose, preparation method and applications patent application matters.The culture medium is based on tobacco material mixture, wherein added with not less than 0 but the carbon source no more than 10% mass fraction and inorganic salts and additive.Prepared bacteria cellulose can partially or completely substitute the cellulose in existing stem, reconstituted tobacco even tobacco leaf in cigarette preparation.Compared with horn of plenty and relatively inexpensive tobacco material or tobacco waste is utilized in the application novelty, it is prepared for the specifically tobacco material culture medium for producing bacteria cellulose using it, in conjunction with more special bacterial strain, fermentation is prepared for the bacterial cellulose product with certain nicotine content.Preliminary Applications show that prepared bacterial cellulose product can directly substitute the application of stem, reconstituted tobacco in prepared by existing cigarette etc., have preferable application value.
Description
Technical field
The invention belongs to technology field of tobacco preparation, and in particular to a kind of bacteria cellulose, preparation method and applications are special
Profit application matters.
Background technology
Bacteria cellulose is a kind of high purity cellulose product of bacterial fermentation production, because it is with excellent characteristic, quilt
It is widely used in including the fields such as medical treatment, papermaking and food.
Raw material currently used for producing bacteria cellulose includes mainly two class of synthetic media and natural medium.Synthesis training
Base is supported based on the ingredients such as sucrose, glucose, cost is higher, and commercial Application is relatively limited.Natural medium is by coconut water, macaque
Peach, grape, watermelon juice and synanthrin etc. are prepared, and Material Cost is cheap, thus are main preparation methods.In addition, having been reported that
Using enzymatic treatment corncob or stalk for producing bacteria cellulose.In short, since natural material amount is big and price is excellent honest and clean,
Reducing has apparent advantage in fermented-producing bacteria fiber cost, thus with the hot culture medium in day come to prepare bacteria cellulose be existing
Row main production process.
In leaf tobacco production, the annual yield of tobacco in China is more than 2,000,000 tons, wherein 25% tobacco leaf is for various reasons, finally
Cigarette industry is not used to directly to produce.In addition, also will produce a large amount of offal in tobacco processing, offal, tobacco rod, crude opium, giving up
Abandon the tobacco wastes such as cigarette.In general, produced tobacco does not utilize resource compared with horn of plenty in tobacco preparation process, except being partly used for
Reconstituted tobacco production is outer, still has a large amount of tobacco resources to be underutilized, and has larger value of exploiting and utilizing.
In tobacco component, key component is the multiple nutritional components such as carbohydrate, nitrogenous class, inorganic salts and organic acid, is carried
It takes object pH value of solution generally also between 5 ~ 6, can be used as natural microculture culture medium without complex process.But
In the prior art, there is not yet using tobacco material as fermentation culture medium for microbe, it is thin that tobacco material is especially used as production
The related report of fungin.
Invention content
The application main purpose be to provide it is a kind of using tobacco material as the bacterial cellulose product prepared by culture medium, should
Bacterial cellulose product contains a certain amount of nicotine component, and the stem, reconstituted tobacco when can preferably be prepared as cigarette even pass
The substitute of system cigarette.
Details are as follows for the technical solution that the application is taken.
A kind of bacteria cellulose preparation tobacco material culture medium, based on tobacco material mixture, wherein adding
Have not less than 0 but the carbon source no more than 10% mass fraction and inorganic salts and additive, pH 4 ~ 7；
The carbon source is one or more of sucrose, glucose, pectin etc. arbitrary proportion mixture, total addition level（Quality point
Number）Preferably 4 ~ 6%（Specific is, for example, 5%）；
The inorganic salts are：One or two kinds of arbitrary proportion mixtures, total addition level in magnesium sulfate, ferrous sulfate（Mass fraction）
Preferably 0.1 ~ 0.2%（Specific is, for example, 0.15%）；
The additive is：One or two kinds of arbitrary proportion mixtures, total addition level in ethyl alcohol, acetic acid（Mass fraction）Preferably
0.2~1.0%（Specific is, for example, 0.5%）；
The tobacco material is：Tobacco leaf, offal, cigarette foam, tobacco rod, offal, scrap one kind in the tobacco materials such as cigarette, flue dust soil or
Several arbitrary proportion mixtures；
The tobacco material mixture is：In mass ratio, 1 ~ 20 times of deionized water of tobacco material quality, 45 DEG C of conditions are added
10 min of lower stirring.
Using bacteria cellulose preparation bacteria cellulose prepared by tobacco material culture medium, make as follows
It is standby to obtain：
（1）Seed liquor is prepared, for one ring of oese picking bacterial strain inclined-plane in seed liquid culture medium, prepared by 28 ~ 30 DEG C of 48 h of culture
Seed liquor；
Seed liquid culture medium：5 g of polyprotein peptone, 5 g of yeast extract, 5 g of glucose, mannitol 5 g, 7H2O·
MgSO41 g, deionized water is settled to 1 L, and after sterilizing, 5 mL absolute ethyl alcohols are added；
The bacterial strain is：Acetobacter xylinum category, Agrobacterium, rhizobium, Aerobacter, alcaligenes etc. can be in tobaccos
Middle production, and the microorganism of bacteria cellulose can be generated；
The acetobacter xylinum that specifically such as preserving number is ATCC23767（Acetobacter xylinum）；
（2）Fermentation prepares bacteria cellulose, by 5 ~ 10% volume ratios, by step（1）Middle seed liquor is inoculated in tobacco material culture medium
In, 8 ~ 18 days are stood at 28 DEG C ~ 30 DEG C, collects the bacteria cellulose film that media surface fermentation generates, deionized water first washs 1
~ 2 times, then washed 3 ~ 4 times with 0.1% NaOH, it is finally cleaned repeatedly with deionized water to pH 7.0 or so, vacuum freeze drying
It is bacteria cellulose finished product afterwards；
In prepared bacteria cellulose, nicotine content is 0.01% ~ 1% mass fraction.
Prepared bacteria cellulose is applied in cigarette preparation, can partially or completely substitute existing stem, reconstituted tobacco
Cellulose even in tobacco leaf.
Compared with horn of plenty and relatively inexpensive tobacco material or tobacco waste is utilized in the application novelty, utilizes its system
For specifically being used to produce the tobacco material culture medium of bacteria cellulose, in conjunction with more special bacterial strain, fermentation is prepared for having
There is the bacterial cellulose product of certain nicotine content.Preliminary Applications show that prepared bacterial cellulose product can be replaced directly
The application of stem, reconstituted tobacco in being prepared for existing cigarette etc. has preferable application value.
Specific implementation mode
Explanation is further explained to the application with reference to embodiment, before introducing specific embodiment, with regard to following realities
Part Experiment background involved in example is applied to be briefly discussed below.
Tobacco material：Tobacco material employed in following embodiments be production of cigarettes during tobacco leaf, offal, cigarette foam,
Tobacco rod, offal scrap the tobacco wastes such as cigarette, flue dust soil, are provided by China Tobacco Henan Industrial Co., Ltd；
Fermentation strain：The Acetobacter xylinus strain of preserving number ATCC23767 employed in following embodiments can be disclosed for one kind and be obtained
Obtain bacterial strain；
Nicotine detects in following embodiments, is detected using high performance liquid chromatography.
Embodiment 1
The there is provided bacteria cellulose preparation tobacco material culture medium of the present embodiment, based on the aqueous solution of tobacco material,
When preparation, after tobacco material is crushed and adds water, 10 min are stirred under the conditions of 40 DEG C, are then allowed to stand 2 h and are naturally cooling to room
Temperature；The glucose of mass fraction 5%, 0.15% magnesium sulfate are added, 0.5% absolute ethyl alcohol is uniformly mixed.
The tobacco material, by quality ratio, tobacco material：Sterile distilled water=1:8；
The tobacco material is：Tobacco leaf, cigarette foam, tobacco rod, scraps the tobacco materials mixtures such as cigarette, flue dust soil at offal, before use,
Grind into powder.
Using bacteria cellulose preparation bacteria cellulose prepared by tobacco material culture medium, make as follows
It is standby to obtain：
（1）Seed liquor is prepared, bacterial strain is accessed in seed liquid culture medium, 30 DEG C of 48 h of culture prepare seed liquor；
Seed culture medium：5 g of polyprotein peptone, 5 g of yeast extract, 5 g of glucose, mannitol 5 g, 7H2O·
MgSO41 g, deionized water is settled to 1 L, and after sterilizing, 5 mL absolute ethyl alcohols are added.
（2）Fermentation prepares bacteria cellulose, by 10% volume ratio, by step（1）Middle seed liquor is inoculated in tobacco material culture
In base, 18 days are stood at 30 DEG C, collects the bacteria cellulose film product that media surface fermentation generates；
Deionized water is first washed 1 ~ 2 time after collected bacteria cellulose film clear water is washed, then washs 3 ~ 4 with 0.1% NaOH
It is secondary, it is finally cleaned repeatedly with deionized water to pH 7.0 or so, is bacteria cellulose finished product after vacuum freeze drying.
In prepared bacteria cellulose, detection shows that nicotine content is 1.1% mass fraction.
Embodiment 2
The preparation method is the same as that of Example 1 with bacteria cellulose for tobacco material culture medium provided herein, and only adjustment member parameter is such as
Under：
Prepared by bacteria cellulose uses tobacco material culture medium, and the tobacco material is offal and scraps cigarette offal 1:1 mass ratio
Mixture, tobacco material：Sterile distilled water=1:10；10min is stirred under the conditions of 45 DEG C；The sulfuric acid for adding 0.05% mass fraction is sub-
Iron adds the absolute ethyl alcohol of 0.5% mass fraction.
When preparing bacteria cellulose, step（2）In, stand 15 days at 28 DEG C.
In prepared bacteria cellulose, detection shows that nicotine content is 0.7% mass fraction.
Embodiment 3
The preparation method is the same as that of Example 1 with bacteria cellulose for tobacco material culture medium provided herein, and only adjustment member parameter is such as
Under：
Prepared by bacteria cellulose uses tobacco material culture medium, and the tobacco material is cigarette primary processing and rolls up what packet workshop was collected
1:The flue dust soil and offal powder of 3 mass ratioes, tobacco material：Sterile distilled water=1:10；10min is stirred under the conditions of 45 DEG C；Addition
The ferrous sulfate of 0.05% mass fraction adds the absolute ethyl alcohol of 4% mass fraction；8% mass fraction sucrose of additional addition is as carbon
Source.
When preparing bacteria cellulose, step（2）In, it is inoculated with seed liquor by 12% volume ratio, 18 days are stood at 28 DEG C.
In prepared bacteria cellulose, detection shows that nicotine content is 0.05% mass fraction.
Embodiment 4
The preparation method is the same as that of Example 1 with bacteria cellulose for tobacco material culture medium provided herein, and only adjustment member parameter is such as
Under：
Prepared by bacteria cellulose uses tobacco material culture medium, and the tobacco material is the tobacco rod that 80-200 mesh crushes, tobacco material：
Sterile distilled water=1:15；40 min are stirred under the conditions of 85 DEG C；
The tobacco leaf powder for adding 5% stirs 5 min, stands cooling.
When preparing bacteria cellulose, step（2）In, it is inoculated with seed liquor by 12% volume ratio, 18 days are stood at 30 DEG C.
In prepared bacteria cellulose, detection shows that nicotine content is 0.08% mass fraction.
Embodiment 5
The preparation method is the same as that of Example 1 for bacteria cellulose provided herein, and tobacco material culture medium and partial parameters adjustment are as follows：
Prepared by bacteria cellulose uses tobacco material culture medium, and the tobacco material is that mass ratio is 6:4 offal is mixed with offal
Object, tobacco material：Sterile distilled water=1:8；15min is stirred under the conditions of 50 DEG C；Filtering is extracted with filter cloth, collects leaching liquor, repeatedly
After three times, the leaching liquor of collection is merged, 0.15% magnesium sulfate, 0.5% absolute ethyl alcohol are added in leaching liquor.
When preparing bacteria cellulose, step（2）In, it is inoculated with seed liquor by 8% volume ratio, 12 d are stood at 30 DEG C.
In prepared bacteria cellulose, detection shows that nicotine content is 0.80% mass fraction.
Embodiment 6
The preparation method is the same as that of Example 1 for bacteria cellulose provided herein, and tobacco material culture medium and partial parameters adjustment are as follows
Prepared by bacteria cellulose uses tobacco material culture medium, and the tobacco material is that mass ratio is 6:4 offal and offal mixture, are used
15 min of ethyl alcohol soak extraction of 10 times of quality, after with filter cloth extract filtering, collect leaching liquor, repeatedly for three times after, by collection
Leaching liquor merges, and after rotary evaporation removes ethyl alcohol, distilled water to original volume is added to obtain the aqueous solution of ethanol extract；
0.15% magnesium sulfate, 9% glucose are added in leaching liquor.
When preparing bacteria cellulose, step（2）In, it is inoculated with seed liquor by 5% volume ratio, 10 days are stood at 30 DEG C.
In prepared bacteria cellulose, detection shows that nicotine content is 0.50% mass fraction.
In general, the application more fully and complete is utilized that raw material sources are abundant and low-cost tobacco is former
Material or tobacco waste, have widened bacteria cellulose raw materials for production range, can more effectively reduce bacterial cellulose product
Production cost.And since the tobaccos such as nicotine endemic element can be wrapped, be adsorbed in bacteria cellulose finished product, to final
It obtains unique properties, with the bacterial cellulose product compared with high added value, has preferably in fact for improving cigarette product quality
With value and promote and apply meaning.
Claims (8)
1. prepared by a kind of bacteria cellulose use tobacco material culture medium, which is characterized in that based on tobacco material mixture,
It is wherein added with not less than 0 but the carbon source no more than 10% mass fraction and inorganic salts and additive, pH 4 ~ 7；
The carbon source is one or more of sucrose, glucose, pectin etc. arbitrary proportion mixture；
The inorganic salts are：One or two kinds of arbitrary proportion mixtures in magnesium sulfate, ferrous sulfate；
The additive is：One or two kinds of arbitrary proportion mixtures in ethyl alcohol, acetic acid；
The tobacco material is：Tobacco leaf, offal, cigarette foam, tobacco rod, to scrap one or more of cigarette, flue dust soil arbitrary proportion mixed
Close object；
The tobacco material mixture is：In mass ratio, the water of 1 ~ 20 times of tobacco material quality is added.
2. prepared by bacteria cellulose as described in claim 1 use tobacco material culture medium, which is characterized in that carbon source total addition level is
4~6%。
3. prepared by bacteria cellulose as described in claim 1 use tobacco material culture medium, which is characterized in that inorganic salts are total addition
Amount is 0.1 ~ 0.2%.
4. prepared by bacteria cellulose as described in claim 1 use tobacco material culture medium, which is characterized in that additive total addition level
It is 0.2 ~ 1.0%.
5. preparing the bacterial fibers prepared by tobacco material culture medium using any one of claim 1 ~ 4 bacteria cellulose
Element, which is characterized in that acquisition is made by the steps：
（1）Seed liquor is prepared, for inoculating strain in seed liquid culture medium, 28 ~ 30 DEG C of cultures prepare seed liquor；
The bacterial strain is：It can be in tobacco in acetobacter xylinum category, Agrobacterium, rhizobium, Aerobacter, alcaligenes
Middle production, and the microorganism of bacteria cellulose can be generated；
（2）Fermentation prepares bacteria cellulose, by 5 ~ 10% volume ratios, by step（1）Middle seed liquor is inoculated in tobacco material culture medium
In, 8 ~ 18 days are stood at 28 DEG C ~ 30 DEG C, the bacteria cellulose film that media surface fermentation generates is collected, is after washing, drying
Bacteria cellulose finished product.
6. bacteria cellulose as claimed in claim 5, which is characterized in that step（1）In, the seed liquid culture medium：Poly egg
White peptone 5 g, 5 g of yeast extract, 5 g of glucose, mannitol 5 g, 7H2O·MgSO41 g, deionized water are settled to 1
After sterilizing, 5 mL absolute ethyl alcohols are added in L.
7. bacteria cellulose as claimed in claim 5, which is characterized in that step（1）In, the bacterial strain is that preserving number is
The acetobacter xylinum of ATCC23767.
8. any one of claim 5 ~ 8 bacteria cellulose is applied in cigarette preparation, which is characterized in that partially or completely replace
For the cellulose in existing stem, reconstituted tobacco or tobacco leaf.
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| CN201810270577.2A CN108642104B (en) | 2018-03-29 | 2018-03-29 | Bacterial cellulose, preparation method and application thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110432533A (en) * | 2019-08-22 | 2019-11-12 | 河南中烟工业有限责任公司 | A kind of heating using micro-wave drying method preparation is not burnt tobacco slice |
| CN110432535A (en) * | 2019-08-22 | 2019-11-12 | 河南中烟工业有限责任公司 | A kind of buccal cigarette of tobacco extract fermentation preparation |
| CN110432534A (en) * | 2019-08-22 | 2019-11-12 | 河南中烟工业有限责任公司 | A kind of preparation method heating tobacco slice of not burning |
| CN111269953A (en) * | 2020-04-27 | 2020-06-12 | 武汉纺织大学 | Method for preparing bacterial cellulose by using waste polyester fabric |
| CN111826994A (en) * | 2020-07-13 | 2020-10-27 | 河南中烟工业有限责任公司 | Preparation method of cigar-flavor cigarette paper |
| CN112056609A (en) * | 2020-09-10 | 2020-12-11 | 河南中烟工业有限责任公司 | Production method of heating cigarette paper-making method sheet |
| CN113528376A (en) * | 2021-06-16 | 2021-10-22 | 河南中烟工业有限责任公司 | Acetobacter gluconicum and application thereof |
| CN115678919A (en) * | 2021-07-26 | 2023-02-03 | 中国科学院青岛生物能源与过程研究所 | Tobacco culture medium, preparation thereof and method for producing bio-based chemicals through direct fermentation |
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| US5327917A (en) * | 1990-08-15 | 1994-07-12 | R. J. Reynolds Tobacco Company | Method for providing a reconstituted tobacco material |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110432533A (en) * | 2019-08-22 | 2019-11-12 | 河南中烟工业有限责任公司 | A kind of heating using micro-wave drying method preparation is not burnt tobacco slice |
| CN110432535A (en) * | 2019-08-22 | 2019-11-12 | 河南中烟工业有限责任公司 | A kind of buccal cigarette of tobacco extract fermentation preparation |
| CN110432534A (en) * | 2019-08-22 | 2019-11-12 | 河南中烟工业有限责任公司 | A kind of preparation method heating tobacco slice of not burning |
| CN111269953A (en) * | 2020-04-27 | 2020-06-12 | 武汉纺织大学 | Method for preparing bacterial cellulose by using waste polyester fabric |
| CN111826994A (en) * | 2020-07-13 | 2020-10-27 | 河南中烟工业有限责任公司 | Preparation method of cigar-flavor cigarette paper |
| CN112056609A (en) * | 2020-09-10 | 2020-12-11 | 河南中烟工业有限责任公司 | Production method of heating cigarette paper-making method sheet |
| CN113528376A (en) * | 2021-06-16 | 2021-10-22 | 河南中烟工业有限责任公司 | Acetobacter gluconicum and application thereof |
| CN113528376B (en) * | 2021-06-16 | 2022-07-26 | 河南中烟工业有限责任公司 | Acetobacter gluconicum and application thereof |
| CN115678919A (en) * | 2021-07-26 | 2023-02-03 | 中国科学院青岛生物能源与过程研究所 | Tobacco culture medium, preparation thereof and method for producing bio-based chemicals through direct fermentation |
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