CN108004184B - Bacillus and method for producing isovaleric acid by using same - Google Patents
Bacillus and method for producing isovaleric acid by using same Download PDFInfo
- Publication number
- CN108004184B CN108004184B CN201810009043.4A CN201810009043A CN108004184B CN 108004184 B CN108004184 B CN 108004184B CN 201810009043 A CN201810009043 A CN 201810009043A CN 108004184 B CN108004184 B CN 108004184B
- Authority
- CN
- China
- Prior art keywords
- bacillus
- isovaleric acid
- tobacco
- strain
- cgmcc
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- 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
-
- 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
- C12N1/02—Separating microorganisms from their culture media
-
- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
Landscapes
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to bacillus and a method for producing isovaleric acid by using the same. The bacillus (A), (B) and (C)Bacillus) Is obtained by separating from tobacco leaves in the middle stage of alcoholization through a selective culture medium, and is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms (CGMCC No. 14609) in 2017, 9 and 13. The strain can synthesize isovaleric acid by taking tobacco industrial waste as a raw material, the isovaleric acid is colorless viscous liquid, and the isovaleric acid has sweet fruit flavor and Vaccinium uliginosum-like flavor after being highly diluted. The fermentation medium used in the process of synthesizing the isovaleric acid by using the bacillus is low in cost, can treat a large amount of waste generated in the tobacco industry, has positive effects on protecting the environment and reasonably utilizing resources, and has good application prospects.
Description
Technical Field
The invention belongs to the technical field of biology, particularly relates to bacillus and application thereof, and particularly relates to the bacillus obtained by screening and a method for producing isovaleric acid by utilizing the strain through fermentation.
Background
Isovaleric acid, also known as 3-methylbutyric acid, is a colorless viscous liquid with a melting point of-29.3 ℃ and a boiling point of 176.5 ℃, and can be mutually dissolved with alcohol, ether, chloroform and the like.
Isovaleric acid has an irritating rancid flavour, a sweet fruity flavour after high dilution, and a vaccinium uliginosum-like flavour. The isovaleric acid is naturally present in valerian oil, vanilla oil, hop oil, bay leaf oil, spearmint oil and the like, and is a food flavor allowed to be used in national standards. The isovaleric acid is used for preparing cheese essence and cream essence and is used for preparing fruit essence in small amount. It can also be used for preparing medicines, spices, seasonings, etc. It can also be used for baking food and meat product. At present, isovaleric acid is mainly used for producing the sedative hypnotic bromoisoylurea. But more are used for the production of fragrances. The isovalerate esters which can be used as perfume are mainly hexyl isovalerate, propyl isovalerate, isoamyl isovalerate, geranyl isovalerate, benzyl isovalerate and cinnamyl isovalerate. The lower isovalerate can be used as edible perfume, and the higher isovalerate can be used in cosmetics.
The current method for industrially producing the isovaleric acid is to oxidize isoamyl alcohol or isovaleraldehyde to obtain the isovaleric acid. The catalyst is mainly used, isoamyl alcohol or isovaleraldehyde is used as a raw material and is prepared through oxidation reaction, and the method not only needs the catalyst, but also has complicated reaction steps and long time. The biosynthesis of isovaleric acid has been reported, probably because the microorganisms for synthesizing isovaleric acid have not been explored or because the microorganisms have low efficiency for synthesizing isovaleric acid, so that the application of isovaleric acid biosynthesis in industry is limited. Therefore, how to overcome the defects of the prior art and improve the application of the microorganism in the production of the isovaleric acid is a problem which needs to be solved in the field of biotechnology at present.
Disclosure of Invention
The invention aims to solve the defects of the prior art and provide bacillus and a method for producing isovaleric acid by utilizing tobacco industry waste, wherein the strain does not need to be transformed, and is easy to separate and culture.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a Bacillus (A), (B) and (C)Bacillus) And the strain has been preserved in China general microbiological culture Collection center (CGMCC) on 9 and 13 months in 2017, and the preservation number is CGMCC 14609.
Bacillus of the present invention: (Bacillus) The preparation method comprises the following steps:
1) separation medium and culture conditions:
diluting an LB liquid culture medium by 8-12 times: 1000mL of distilled water; tryptone 1 g; yeast extract 0.5 g; 1g of NaCl; sterilizing at 121 deg.C for 20 min with natural pH;
2) strain screening: adding agar of 20g/L into the liquid culture medium to prepare a solid culture medium for strain screening;
3) the separation method comprises the following steps:
randomly extracting 2g of tobacco leaf samples in the middle stage of alcoholization, cutting into pieces under aseptic condition, soaking in 150mL of sterile physiological saline, performing shaking culture for 60 minutes under the conditions of 37 ℃ and 180 r/min of rotation speed, filtering with sterile single-layer gauze, taking filtrate, centrifuging, discarding supernatant, re-suspending with 5mL of sterile water to obtain original bacterial suspension, and diluting 100 mu L of original bacterial solution to 10 mu L respectively0，10-1，10-2，10-3，10-4，10-5And respectively sucking 200 mu L of the bacillus subtilis and coating the bacillus subtilis on an LB culture medium diluted by 8-12 times, carrying out inverted culture at 37 ℃ for 24 hours, and purifying the cultured bacterial colonies by adopting a plate marking method until single bacillus colonies are obtained.
Using Bacillus bacteria (A), (B)Bacillus) The method for producing the isovaleric acid comprises the following steps: inoculating the bacillus suspension into a fermentation culture medium according to the inoculation amount of 5-10%, placing the bacillus suspension into a shaking table, culturing and fermenting for 5 days at 37 ℃, and extracting by ethyl acetate to obtain isovaleric acidDetection was by GC-MS.
The fermentation medium is a culture medium taking tobacco industrial waste as nutrients and comprises the following components: 20g/L of glucose, 5g/L of tobacco industrial waste and 7.0-7.4 of pH value.
The tobacco industry waste is one or a combination of more of tobacco stems, tobacco fragments, tobacco powder and cigarette ash rods.
The shaker was 140 rpm.
The invention relates to bacillus for producing isovaleric acid by using tobacco industrial waste as raw material (B)Bacillus) Is obtained by separating from tobacco leaves in the middle stage of alcoholization through selective culture medium.
The bacillus is gram-positive bacillus, has spores, produces nearly-neutral elliptic spores, expands cysts, and has the size of 0.5 mu mx (1.0-1.5) mu m observed under a microscope. The somatic cells are short rod-shaped. On the surface of an LB solid medium, the thalli form semitransparent milky microcolonies, the surfaces of the microcolonies are moist and smooth, and the edges of the microcolonies are neat. The 16SrDNA sequence of the strain obtained by the general primer PCR has the highest similarity with the 16SrDNA sequence of the rhizobium M-5 (shown in figure 1).
Wherein the PCR upstream primer is 27F: 5'-agagtttgatcctggctcag-3' (SEQ ID NO. 2);
the downstream primer is 1492R: 5'-cggctaccttgttacgactt-3' (SEQ ID NO. 3).
The method provides a new source of isovaleric acid, the strain of the invention does not need to be transformed, and is easy to separate and culture, besides, the used fermentation medium has low cost, and can treat a large amount of waste generated in the tobacco industry, which has positive effects on environmental protection and reasonable resource utilization.
Compared with the prior art, the invention has the beneficial effects that:
1) the bacillus obtained by screening can utilize the tobacco industry waste as a main nutrient source, can treat a large amount of waste generated in the tobacco industry, and has positive effects on protecting the environment and reasonably utilizing resources.
2) The bacillus can ferment tobacco industrial wastes to produce isovaleric acid. The isovaleric acid is a colorless viscous liquid, and has sweet fruit flavor and Vaccinium uliginosum-like flavor after being highly diluted. The method can be used for producing spices mainly used for preparing cheese and cream essence and trace fruit essence on one hand, and can also be used for producing the sedative hypnotic bromoisovaleryl urea on the other hand.
3) The invention provides a new approach for synthesizing isovaleric acid, the strain of the strain is not required to be modified, and is easy to separate and culture, so that the problem of producing isovaleric acid by a biological method is expected to be solved, and the industrial production and the application of isovaleric acid are promoted.
Drawings
FIG. 1 is a phylogenetic tree of Bacillus bacteria selected in the present invention;
FIG. 2 is a mass spectrogram of isovaleric acid obtained by fermentation according to the present invention;
bacillus (A), (B)Bacillus) The microbial inoculum is preserved in the China general microbiological culture Collection center (CGMCC) at 13.9.2017 with the preservation number of CGMCC number 14609, and the preservation address is No.3 of Xilu 1 of Beijing university facing Yang, China academy of sciences.
Detailed Description
The present invention will be described in further detail with reference to examples.
It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples do not specify particular techniques or conditions, and are performed according to the techniques or conditions described in the literature in the art or according to the product specifications. The materials or instruments used are not indicated by manufacturers, and are all conventional products available by purchase.
EXAMPLE 1 isolation of microorganisms for producing Isovaleric acid Using tobacco Industrial waste
1. Separation medium and culture conditions:
10-fold (or 8-fold, or 12-fold) dilution of LB liquid medium: 1000mL of distilled water; tryptone 1 g; yeast extract 0.5 g; 1g of NaCl; natural pH and sterilization;
the above medium was sterilized at 121 ℃ for 20 minutes.
20g/L agar was added to the above medium to prepare a solid medium for strain screening.
2. The separation method comprises the following steps:
randomly extracting 2g of tobacco leaf samples in the middle stage of alcoholization, cutting into pieces under aseptic condition, soaking in 150mL of sterile physiological saline, performing shaking culture for 60 minutes under the conditions of 37 ℃ and 180 r/min of rotation speed, filtering with sterile single-layer gauze, taking filtrate, centrifuging, discarding supernatant, re-suspending with 5mL of sterile water to obtain original bacterial suspension, and diluting 100 mu L of original bacterial solution to 10 mu L respectively0，10-1，10-2，10-3，10-4，10-5And respectively sucking 200. mu.L of LB culture medium diluted 10 times (or 8 times, or 12 times), performing inverted culture at 37 ℃ for 24 hours, and purifying the cultured colonies by using a plate-streaking method until single colonies are obtained.
Example 2 identification of species of isolated microorganisms
(1) And (3) culturing the bacteria separated from the selected culture medium in LB liquid culture medium diluted by 10 times, and obtaining bacterial suspension of the corresponding strain after 12 hours.
(2) Taking 500 mu L of bacterial suspension, centrifuging, removing supernatant, extracting genome DNA, and performing Polymerase Chain Reaction (PCR) by taking the extracted genome DNA as a template.
The PCR upstream primer is 27F: 5'-agagtttgatcctggctcag-3' (SEQ ID NO. 2);
the downstream primer is 1492R: 5'-cggctaccttgttacgactt-3' (SEQ ID NO. 3).
The PCR reaction system and procedure were as follows:
PCR amplification System (25. mu.L):
PCR amplification procedure:
30 cycles.
(3) 5 mu L of PCR reaction solution and DNA marker 2000 are respectively taken and gel electrophoresis is applied to carry out PCR product verification.
(4) PCR products with fluorescence bands appearing at about 1600bp are sent to a sequencing company for sequencing, and after the sequencing results are spliced, the 16SrDNA sequence has 99% similarity with the 16SrDNA sequence of the bacillus M-5 and is input into BLSAT (https:// blast. The bacillus is gram-positive bacillus, has spores, produces nearly-neutral elliptic spores, expands cysts, and has the size of 0.5 mu mx (1.0-1.5) mu m observed under a microscope. The somatic cells are short rod-shaped. On the surface of an LB solid medium, the thalli form semitransparent milky microcolonies, the surfaces of the microcolonies are moist and smooth, and the edges of the microcolonies are neat. The strain is preliminarily identified to be the rhizopus M-5, the strain is preserved to the China general microbiological culture Collection center, and the preservation number is as follows: CGMCC number 14609. The sequencing result is shown as SEQ ID NO. 1.
Example 3 production of Isovaleric acid from tobacco industry waste
The culture medium taking the tobacco industry waste as the nutrient comprises the following components: 20g/L of glucose, 5g/L of tobacco industrial waste (tobacco stem/fragment: tobacco powder is 50:30: 20), and pH value is 7.0-7.4;
under aseptic conditions, bacillus was inoculated into sterilized LB liquid medium and cultured until OD600 was 1. The above-mentioned strain was inoculated in a 250mL shake flask containing 50mL of a sterilized tobacco industrial waste nutrient medium at an inoculum size of 3%, a control group not inoculated with the strain was placed, and the strain was cultured in a shaker at 140 rpm at 37 ℃ for 5 days, followed by detection by GC-MS.
Qualitative detection of isovaleric acid: centrifuging the fermentation liquor at 12000 rpm for 10 min, removing precipitate, extracting the supernatant with ethyl acetate to obtain ethyl acetate extract, removing part of solvent by rotary evaporation, removing water with anhydrous sodium sulfate, filtering with 0.22 μm membrane, and performing qualitative detection by GC-MS. An Innovax capillary chromatographic column (30 m x 0.25mm, 0.25 μm.) was used, the injection port temperature was 250 ℃, the ion source temperature was 230 ℃, the temperature program was 40 ℃ for 6 minutes, then the temperature program was 3 ℃/minute to 100 ℃, and then the temperature program was 5 ℃/minute to 230 ℃. The mass spectrum obtained is shown in figure 1, and is compared with a standard library to obtain isovaleric acid.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Sequence listing
<110> tobacco industry Limited liability company in Yunnan
<120> a bacillus and a method for producing isovaleric acid by using the same
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1402
<212> DNA
<213> Bacillus
<400> 1
cttcggcggt ggctcataaa ggttacctca ccgacttcgg gtgttgcaaa ctctcgtggt 60
gtgacgggcg gtgtgtacaa ggcccgggaa cgtattcacc gcggcatgct gatccgcgat 120
tactagcgat tccagcttca cgcagtcgag ttgcagactg cgatccgaac tgagaacaga 180
tttatgggat tggctaaacc ttgcggtctt gcagcccttt gttctgtcca ttgtagcacg 240
tgtgtagccc aggtcataag gggcatgatg atttgacgtc atccccacct tcctccggtt 300
tgtcaccggc agtcacctta gagtgcccaa ctgaatgctg gcaactaaga tcaagggttg 360
cgctcgttgc gggacttaac ccaacatctc acgacacgag ctgacgacaa ccatgcacca 420
cctgtcactc tgtccccgaa gggaaagccc tatctctagg gttgtcagag gatgtcaaga 480
cctggtaagg ttcttcgcgt tgcttcgaat taaaccacat gctccaccgc ttgtgcgggc 540
ccccgtcaat tcctttgagt ttcagtcttg cgaccgtact ccccaggcgg agtgcttaat 600
gcgttagctg cagcactaag gggcggaaac cccctaacac ttagcactca tcgtttacgg 660
cgtggactac cagggtatct aatcctgttc gctccccacg ctttcgctcc tcagcgtcag 720
ttacagacca gagagtcgcc ttcgccactg gtgttcctcc acatctctac gcatttcacc 780
gctacacgtg gaattccact ctcctcttct gcactcaagt ttcccagttt ccaatgaccc 840
tccccggttg agccgggggc tttcacatca gacttaagaa accgcctgcg agccctttac 900
gcccaataat tccggacaac gcttgccacc tacgtattac cgcggctgct ggcacgtagt 960
tagccgtggc tttctggtta ggtaccgtca aggtgcgagc agttactctc gcacttgttc 1020
ttccctaaca acagagcttt acgatccgaa aaccttcatc actcacgcgg cgttgctccg 1080
tcagactttc gtccattgcg gaagattccc tactgctgcc tcccgtagga gtctgggccg 1140
tgtctcagtc ccagtgtggc cgatcaccct ctcaggtcgg ctacgcatcg tcgccttggt 1200
gagccattac cccaccaact agctaatgcg ccgcgggtcc atctgtaagt gacagccgaa 1260
accgtctttc atccttgaac catgcggttc aaggaactat ccggtattag ctccggtttc 1320
ccggagttat cccagtctta caggcaggtt acccacgtgt tactcacccg tccgccgcta 1380
acatccggga gcaagctccc tt 1402
<210> 2
<211> 20
<212> DNA
<213> Artificial sequence ()
<400> 2
agagtttgat cctggctcag 20
<210> 3
<211> 20
<212> DNA
<213> Artificial sequence ()
<400> 3
cggctacctt gttacgactt 20
Claims (4)
1. A Bacillus (A), (B) and (C)Bacillus) And the strain has been preserved in China general microbiological culture Collection center (CGMCC) on 9 and 13 months in 2017, and the preservation number is CGMCC 14609.
2. The Bacillus of claim 1 (Bacillus) The application in the production of isovaleric acid is characterized by comprising the following steps: inoculating the bacillus suspension into a fermentation culture medium according to the inoculation amount of 5-10%, placing the bacillus suspension into a shaking table, culturing for 5 days at 37 ℃, and extracting by ethyl acetate to obtain isovaleric acid; the fermentation medium is a culture medium taking tobacco industrial waste as nutrients, and comprises the components of 20g/L of glucose, 5g/L of tobacco industrial waste and 7.0-7.4 of pH value.
3. The Bacillus of claim 2 (A)Bacillus) The application of the isovaleric acid in the production of isovaleric acid is characterized in that the tobacco industry waste comprises one or a combination of more of tobacco stems, tobacco fragments, tobacco powder and tobacco ash rods.
4. The Bacillus of claim 2 (A)Bacillus) Use in the production of isovaleric acid, characterized in thatThe shaker of (1) is 140 revolutions per minute.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810009043.4A CN108004184B (en) | 2018-01-04 | 2018-01-04 | Bacillus and method for producing isovaleric acid by using same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810009043.4A CN108004184B (en) | 2018-01-04 | 2018-01-04 | Bacillus and method for producing isovaleric acid by using same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108004184A CN108004184A (en) | 2018-05-08 |
| CN108004184B true CN108004184B (en) | 2021-01-29 |
Family
ID=62049765
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810009043.4A Active CN108004184B (en) | 2018-01-04 | 2018-01-04 | Bacillus and method for producing isovaleric acid by using same |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108004184B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109295114A (en) * | 2018-10-29 | 2019-02-01 | 南方医科大学 | Application of the staphylococcus epidermis in synthesis isobutyric acid |
| CN110776412B (en) * | 2019-11-12 | 2022-04-22 | 万华化学集团股份有限公司 | Method for preparing isovaleric acid, ligand, complex and application thereof in catalytic system |
| CN113774072B (en) * | 2021-07-15 | 2023-05-02 | 湖南省农业科学院 | Citrate synthase gene, citrate synthase, use as target and herbicide |
| CN114350540B (en) * | 2021-11-30 | 2024-04-09 | 浙江中烟工业有限责任公司 | Bacillus and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000020597A1 (en) * | 1998-10-02 | 2000-04-13 | Societe Des Produits Nestle S.A. | Novel strains of the bacillus subtilis group for food fermentation |
| CN105969703A (en) * | 2016-07-27 | 2016-09-28 | 云南中烟工业有限责任公司 | Bacillus licheniformis and application thereof |
| US20170067064A1 (en) * | 2004-06-29 | 2017-03-09 | Basf Se | Gene Products of Bacillus Licheniformis Which Form Odorous Substances and Improved Biotechnological Production Methods Based Thereon |
| CN107418922A (en) * | 2017-09-12 | 2017-12-01 | 云南中烟工业有限责任公司 | A kind of rhizosphere bacillus and its method using tobacco industry discarded object production hexyl acetate |
-
2018
- 2018-01-04 CN CN201810009043.4A patent/CN108004184B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000020597A1 (en) * | 1998-10-02 | 2000-04-13 | Societe Des Produits Nestle S.A. | Novel strains of the bacillus subtilis group for food fermentation |
| US20170067064A1 (en) * | 2004-06-29 | 2017-03-09 | Basf Se | Gene Products of Bacillus Licheniformis Which Form Odorous Substances and Improved Biotechnological Production Methods Based Thereon |
| CN105969703A (en) * | 2016-07-27 | 2016-09-28 | 云南中烟工业有限责任公司 | Bacillus licheniformis and application thereof |
| CN107418922A (en) * | 2017-09-12 | 2017-12-01 | 云南中烟工业有限责任公司 | A kind of rhizosphere bacillus and its method using tobacco industry discarded object production hexyl acetate |
Non-Patent Citations (1)
| Title |
|---|
| 烟叶产香菌的分离及其在烟用香料制备中的应用;吕品等;《烟草化学》;20091231(第258期);37-42 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108004184A (en) | 2018-05-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108004184B (en) | Bacillus and method for producing isovaleric acid by using same | |
| CN106085889B (en) | Pichia guilliermondii 3-J15 and its application | |
| CN107904177B (en) | Dendrobium officinale endophytic fungus strain, exopolysaccharide produced by same, and extraction method and application of exopolysaccharide | |
| CN109722393B (en) | Tobacco yellow soil-borne bacterium | |
| CN103725632B (en) | The subtilis SM that one strain is separated from offal concentrated solution and the application papermaking-method reconstituted tobaccos thereof | |
| CN109797114B (en) | Micrococcus and application thereof in production of 2, 3-butanediol | |
| CN109554318A (en) | Gluconic acid acetobacter and its application in a kind of fermented tea | |
| CN112080435A (en) | Purple lilac spore strain and application thereof in degrading chicken feather | |
| CN110079470B (en) | Pseudomonas with antibacterial activity | |
| US10364446B2 (en) | Streptomyces psammoticus and methods of using the same for vanillin production | |
| CN114540252A (en) | Microbacterium P6 for converting livestock and poultry breeding waste and application | |
| CN110669683A (en) | Saccharomyces cerevisiae and application thereof in preparation of tobacco flavor | |
| CN107418922B (en) | Rhizosphere bacillus and method for producing hexyl acetate by utilizing tobacco industrial waste | |
| CN104498372B (en) | Fusarium oxysporum BM201, its compound pectinase produced and compound pectinase preparation method and application | |
| CN106967612B (en) | Method for producing five ustilaginoidea virens toxins by using ustilaginoidea virens | |
| CN105969703B (en) | A kind of bacillus licheniformis and its application | |
| CN105063016A (en) | Method for extracting microorganism total DNA from yellow rice wine wheat koji | |
| CN109136146B (en) | Tobacco soil bacterium, and separation method and application thereof | |
| CN104073453B (en) | Bacillus amyloliquefaciens and application thereof in control over geosmin smell in white spirit | |
| CN104087526B (en) | A kind of bacillus licheniformis is utilized to control the method for earthy in white wine | |
| CN104004693A (en) | Bacillus pumilus and application of bacillus pumilus in controlling over geosmin in white spirit | |
| CN113308384B (en) | Cordyceps sinensis strain and application thereof | |
| CN114752546A (en) | Method for inducing callus to generate shikimic acid by using aniseed seeds | |
| CN108741208A (en) | One plant of flavouring bacterium detached from reproduced tobacco concentrated liquid and its application | |
| CN107699531A (en) | A kind of hippophae plant Frankia isolated culture method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |