CN109294939A - A method of the antibacterial action of enhancing Bacillus natto metabolite - Google Patents
A method of the antibacterial action of enhancing Bacillus natto metabolite Download PDFInfo
- Publication number
- CN109294939A CN109294939A CN201811015168.4A CN201811015168A CN109294939A CN 109294939 A CN109294939 A CN 109294939A CN 201811015168 A CN201811015168 A CN 201811015168A CN 109294939 A CN109294939 A CN 109294939A
- Authority
- CN
- China
- Prior art keywords
- bacillus natto
- antibacterial action
- metabolite
- seed liquor
- culture medium
- 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.)
- Pending
Links
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
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- 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
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Beans For Foods Or Fodder (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of methods of antibacterial action for enhancing Bacillus natto metabolite, belong to technical field of bioengineering.Bacillus natto was cultivated 8-15h before this by the present invention in seed culture medium, continue in 0 DEG C of low-temperature treatment 24-28h, again in fermentation medium culture 24-36h, the antibacterial action of obtained Bacillus natto metabolite is got well than without the antibacterial action of the metabolite of low-temperature treatment.The method of the present invention is simple, is convenient for operation, can be used for enhancing the antibacterial action of Bacillus natto metabolite.
Description
Technical field
The present invention relates to a kind of methods of antibacterial action for enhancing Bacillus natto metabolite, belong to biotechnology neck
Domain.
Background technique
Preservative used in food industry is chemical preservative mostly, there is certain toxic side effect to human body.Develop day
Right preservative is the hot spot of current food antiseptic research, is found from the metabolite of the microorganism of traditional fermented food natural raw
Object preservative is even more a safely and effectively method.
The metabolite of Bacillus natto has broad-spectrum antibacterial action, has antagonism to the microorganism of common contaminated food products.
It is studied through Japanese expert, the metabolite of Bacillus natto all has certain antibacterial activity to bacterium, saccharomycete and mould, has suppression
The effect of the pathogenic bacteria such as salmonella, typhoid fever bacterium, dysentery bacillus and O-157, H7 Escherichia coli processed.Contain in the mucus of Bacillus natto metabolism
Primary Escherichia coli and salmonella are had stronger antibacterial action by bacitracin, polymyxins and 2,6- pyridine shuttle acid.It is mostly viscous
Rhzomorph is strong to the antibacterial activity of Gram-negative bacteria, and sensitive prime bacterium has Escherichia coli, salmonella, Pasteurella, cloth Lu Shi
Bacterium, vibrios, shigella dysenteriae, Pseudomonas aeruginosa etc. especially have powerful bactericidal effect to Pseudomonas aeruginosa.Gramicidins and bacitracin
Mainly act on gram-positive bacteria.Bacillus skin is effective to resistant Staphylococcus aureus, enterococcus, streptococcus, to conveyor screw
It is also effective with actinomyces but invalid to Gram-negative bacteria.
The antibacterial material that Bacillus natto generates undergoes microbial fermentation and generates, and belongs to pure biological agent, equal to people, animal
Nonhazardous effect, therefore these antibacterial materials can be separated by biologically pure technology, it is anti-for producing natural biological
Rotten agent, in terms of it is used in food additives.Its effect can be equivalent to preservative, and without any toxicity and side effects, be met with this
Requirement of the consumer to antiseptics for natural food.
Summary of the invention
Object of the present invention is to solve the problems, such as that Bacillus natto metabolite bacteriostasis is weaker, a kind of enhancing Bacillus natto generation is provided
The method for thanking to the antibacterial action of product.
Technical solution of the present invention is summarized as follows:
A method of the antibacterial action of enhancing Bacillus natto metabolite includes the following steps:
(1) bafillus natto (Bacillus natto) is seeded in seed culture medium, on the shaking table of 220r/min
37 DEG C of culture 8-15h, obtain seed liquor；
(2) Cold pretreatment: collecting thallus for cultured seed liquor with (4 DEG C) centrifugation 15min of 12000r/min low temperature,
Thallus is resuspended in 0 DEG C of Cold pretreatment 24-48h of progress in fresh seed culture medium.
(3) step (2) pretreated seed liquor is inoculated in fermentation medium by the inoculum concentration of 3%-5%, 37 DEG C
24-36h is cultivated on the shaking table of 220r/min.
Beneficial effects of the present invention:
The present invention obtains the Cold pretreatment of thallus progress 24-48h to 8-15h is cultivated in seed culture medium, and thallus is again
(locate in advance without low temperature in the antibacterial action effect for the metabolite that fermentation medium top fermentation 24-36h is obtained than control group
Reason) it to get well.Method of the invention, it is simple and easy, it can effectively and quickly enhance the antibacterial action of Bacillus natto metabolite.
Specific embodiment
Bafillus natto (Bacillus natto) belongs to bacillus subtilis Pseudomonas, deposit number BNCC185324,
In in June, 2017 structure receive biological (www.bnbio.com) from north, address: the Chaoyang District, Beijing City garden Shuan Hui No. 5 building of first.
Escherichia coli (Escherichia coli), deposit number BNCC336902, in March, 2017 structure it is raw from Bei Na
Object (www.bnbio.com), address: the Chaoyang District, Beijing City garden Shuan Hui No. 5 building of first.
Seed culture medium: peptone 1wt%, beef extract 0.3wt%, NaCl 0.5wt%, glucose 0.5wt%, yeast
Cream 0.5wt%, PH 7.0.
Fermentation medium: glucose 2wt%, peptone 0.5wt%, NaCl 0.5wt%, K2HPO40.4wt%,
KH2PO40.2wt%, pH 7.0.
Embodiment 1 (G1)
A method of the antibacterial action of enhancing Bacillus natto metabolite includes the following steps:
(1) bafillus natto (Bacillus natto) is seeded in above-mentioned seed culture medium, 220r/min's shakes
The upper 37 DEG C of cultures 8h of bed, obtains seed liquor；
(2) Cold pretreatment: by cultured seed liquor with 12000r/min, 4 DEG C of centrifugation 15min collect thallus, by bacterium
Weight is suspended from fresh seed culture medium and carries out 0 DEG C of Cold pretreatment for 24 hours；
(3) step (2) pretreated seed liquor is inoculated in fermentation medium by 3% inoculum concentration, 37 DEG C
It is cultivated for 24 hours on the shaking table of 220r/min.
Reference examples 1 (CK1)
(1) bafillus natto (Bacillus natto) is seeded in seed culture medium, on the shaking table of 220r/min
37 DEG C of culture 8h, obtain seed liquor；
(2) to illumination pretreatment method: cultured seed liquor is collected into thallus with (4 DEG C) centrifugation 15min of 12000r/min,
Thallus is resuspended in fresh seed culture medium and carries out room temperature pretreatment for 24 hours；
(3) step (2) pretreated seed liquor is inoculated in fermentation medium by 3% inoculum concentration, 37 DEG C
It is cultivated for 24 hours on the shaking table of 220r/min.
Take the bacteria suspension of step (3) after fermentation that supernatant, as Bacillus natto metabolite solution is collected by centrifugation；It takes
106~107cfu/m L indicator bacteria (Escherichia coli) suspension 0.2m L is added in 2 plating mediums prepared, and coating is equal
It is even, then 3 sterile Oxford cups are put into every ware with aseptic nipper, it is drawn in 0.2m L embodiment 1 with Sterile pipette respectively
The metabolite solution of CK1 is put into the Oxford cup of respective markers in G1 and reference examples 1, and each antibacterial circle diameter is measured after 48h, is taken
Mean value.(being shown in Table 1)
Embodiment 2 (G2)
A method of the antibacterial action of enhancing Bacillus natto metabolite includes the following steps:
(1) bafillus natto (Bacillus natto) is seeded in seed culture medium, on the shaking table of 220r/min
37 DEG C of culture 12h, obtain seed liquor；
(2) Cold pretreatment: cultured seed liquor is collected into thallus with (4 DEG C) centrifugation 15min of 12000r/min, by bacterium
Weight is suspended from 0 DEG C of Cold pretreatment 36h of progress in fresh seed culture medium；
(3) step (2) pretreated seed liquor is inoculated in fermentation medium by 4% inoculum concentration, 37 DEG C
30h is cultivated on the shaking table of 220r/min.
Reference examples 2 (CK2)
(1) bafillus natto (Bacillus natto) is seeded in seed culture medium, on the shaking table of 220r/min
37 DEG C of culture 12h, obtain seed liquor；
(2) to illumination pretreatment method: cultured seed liquor is collected into thallus with (4 DEG C) centrifugation 15min of 12000r/min,
Thallus is resuspended in fresh seed culture medium and carries out room temperature pretreatment 36h；
(3) step (2) pretreated seed liquor is inoculated in fermentation medium by 4% inoculum concentration, 37 DEG C
30h is cultivated on the shaking table of 220r/min.
Take bacteria suspension after fermentation that supernatant, as Bacillus natto metabolite solution is collected by centrifugation；Take 106~
107cfu/m L indicator bacteria (Escherichia coli) suspension 0.2m L is added in 2 plating mediums prepared, and coating is uniform, then
3 sterile Oxford cups are put into every ware with aseptic nipper, draw the metabolism of 0.2m L G2 and CK2 with Sterile pipette respectively
Reaction mixture is put into the Oxford cup of respective markers, and each antibacterial circle diameter is measured after 48h, takes mean value.(being shown in Table 1)
Embodiment 3 (G3)
A method of the antibacterial action of enhancing Bacillus natto metabolite includes the following steps:
(1) bafillus natto (Bacillus natto) is seeded in seed culture medium, on the shaking table of 220r/min
37 DEG C of culture 15h, obtain seed liquor；
(2) Cold pretreatment: cultured seed liquor is collected into thallus with (4 DEG C) centrifugation 15min of 12000r/min, by bacterium
Weight is suspended from 0 DEG C of Cold pretreatment 48h of progress in fresh seed culture medium；
(3) step (2) pretreated seed liquor is inoculated in fermentation medium by 5% inoculum concentration, 37 DEG C
36h is cultivated on the shaking table of 220r/min.
Reference examples 3 (CK3)
(1) bafillus natto (Bacillus natto) is seeded in seed culture medium, on the shaking table of 220r/min
37 DEG C of culture 15h, obtain seed liquor；
(2) to illumination pretreatment method: cultured seed liquor is collected into thallus with (4 DEG C) centrifugation 15min of 12000r/min,
Thallus is resuspended in fresh seed culture medium and carries out room temperature pretreatment 48h；
(3) step (2) pretreated seed liquor is inoculated in fermentation medium by 5% inoculum concentration, 37 DEG C
36h is cultivated on the shaking table of 220r/min.
Take bacteria suspension after fermentation that supernatant, as Bacillus natto metabolite solution is collected by centrifugation；Take 106~
107cfu/m L indicator bacteria (Escherichia coli) suspension 0.2m L is added in 2 plating mediums prepared, and coating is uniform, then
3 sterile Oxford cups are put into every ware with aseptic nipper, draw the metabolism of 0.2m L G3 and CK3 with Sterile pipette respectively
Reaction mixture is put into the Oxford cup of respective markers, and each antibacterial circle diameter is measured after 48h, takes mean value.(being shown in Table 1)
Table 1
| Experiment numbers | Bacteriostatic diameter (mm) |
| CK1 | 9.2 |
| G1 | 11.6 |
| CK2 | 10.3 |
| G2 | 12.7 |
| CK3 | 10.9 |
| G3 | 13.2 |
As can be seen from Table 1 after 4 DEG C of Cold pretreatments, antibacterial circle diameter increases 26.07% in embodiment 1, in fact
It applies antibacterial circle diameter in example 2 and increases 23.30%, antibacterial circle diameter increases 21.10% in embodiment 3.From above three reality
It applies in example as can be seen that the bacteriostasis of Bacillus natto metabolite is improved after 4 DEG C of Cold pretreatments.
Claims (1)
1. a kind of method for the antibacterial action for enhancing Bacillus natto metabolite, includes the following steps:
(1) bafillus natto (Bacillus natto) is seeded in seed culture medium, 37 DEG C on the shaking table of 220r/min
8-15h is cultivated, seed liquor is obtained；
(2) Cold pretreatment: by cultured seed liquor with 12000r/min, 4 DEG C of low-temperature centrifugation 15min collect thallus, by bacterium
Weight is suspended from 0 DEG C of Cold pretreatment 24-48h of progress in fresh seed culture medium；
(3) step (2) pretreated seed liquor is inoculated in fermentation medium by the inoculum concentration of 3%-5%, 37 DEG C
24-36h is cultivated on the shaking table of 220r/min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811015168.4A CN109294939A (en) | 2018-08-31 | 2018-08-31 | A method of the antibacterial action of enhancing Bacillus natto metabolite |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811015168.4A CN109294939A (en) | 2018-08-31 | 2018-08-31 | A method of the antibacterial action of enhancing Bacillus natto metabolite |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109294939A true CN109294939A (en) | 2019-02-01 |
Family
ID=65165965
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811015168.4A Pending CN109294939A (en) | 2018-08-31 | 2018-08-31 | A method of the antibacterial action of enhancing Bacillus natto metabolite |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109294939A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6016569A (en) * | 1983-07-08 | 1985-01-28 | Osaka Gas Co Ltd | Preparation of fermented soybean |
| JPH067156A (en) * | 1992-06-25 | 1994-01-18 | Gold Kosan Kk | Proliferation and treatment of microorganism producing tacky substance |
-
2018
- 2018-08-31 CN CN201811015168.4A patent/CN109294939A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6016569A (en) * | 1983-07-08 | 1985-01-28 | Osaka Gas Co Ltd | Preparation of fermented soybean |
| JPH067156A (en) * | 1992-06-25 | 1994-01-18 | Gold Kosan Kk | Proliferation and treatment of microorganism producing tacky substance |
Non-Patent Citations (1)
| Title |
|---|
| 纪宁等: "纳豆菌产抗菌物质培养条件的研究", 《中国乳品工业》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Vincent et al. | Antibacterial activity associated with Lactobacillus acidophilus | |
| CN103911306B (en) | Lactobacillus gasseri bacterial strain and application thereof | |
| CN102690771B (en) | Functional lactobacillus plantarum and application thereof | |
| CN103911305B (en) | Lactobacillus crispatus bacterial strain and uses thereof | |
| CN111269865B (en) | A strain of Brevibacterium lateralis S62-9 and use thereof | |
| CN103421715A (en) | Lactobacillus rhamnosus and application thereof | |
| CN111548968A (en) | Bacillus coagulans BC99 and screening method thereof | |
| CN109439577A (en) | One plant of broad-spectrum antiseptic bacillus amyloliquefaciens and its application | |
| CN109679882A (en) | One Enterococcus faecalis DT1-1 and its application | |
| CN104140937A (en) | Lactobacillus plantarum KLDS1001 and application of lactobacillus plantarum KLDS1001 to inhibition of streptococcus mutans | |
| CN107312732A (en) | A kind of probiotic feed additive | |
| CN111647533A (en) | Bacillus subtilis, preparation and application thereof | |
| CN115317522A (en) | Application of lactobacillus rhamnosus R7970 in preparation of pathogenic bacterium inhibition product | |
| CN103911309B (en) | Lactobacillus gasseri bacterial strain and application thereof | |
| CN103911308A (en) | Lactobacillus crispatus strain and its uses | |
| CN108410763B (en) | Bifidobacterium longum TC01 and application and product using same | |
| CN103911307B (en) | Lactobacillus crispatus bacterial strain and uses thereof | |
| CN105385729A (en) | Lactococcus garviea bacteriocin, preparing method thereof and application thereof | |
| CN112725222B (en) | Bacillus subtilis Q3 for producing complex enzyme, culture method and application | |
| CN105018395B (en) | One bacillus pumilus and its application in alternaria leaf spot of apple is prevented | |
| CN110452857B (en) | A Lactobacillus plantarum producing non-protein small molecule antibacterial metabolites and its application | |
| CN109294939A (en) | A method of the antibacterial action of enhancing Bacillus natto metabolite | |
| CN117757682A (en) | Enterococcus faecium and application thereof | |
| CN111567807A (en) | Rhamnose-containing lactobacillus composition for inhibiting inflammatory reaction and resisting vaginitis and application thereof | |
| CN113215067B (en) | Lactobacillus breve CSHRR5-3 strain in VBNC state and its application |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20190201 |
|
| WD01 | Invention patent application deemed withdrawn after publication |