CN110093279A - The manufacturing method of new aspergillus niger A-T1 bacterial strain and the natural antimicrobial substance with this - Google Patents
The manufacturing method of new aspergillus niger A-T1 bacterial strain and the natural antimicrobial substance with this Download PDFInfo
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- CN110093279A CN110093279A CN201910193605.XA CN201910193605A CN110093279A CN 110093279 A CN110093279 A CN 110093279A CN 201910193605 A CN201910193605 A CN 201910193605A CN 110093279 A CN110093279 A CN 110093279A
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- bacterial strain
- aspergillus niger
- flavonoids
- antimicrobial substance
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Classifications
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- C—CHEMISTRY; METALLURGY
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- 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
- C12N1/14—Fungi; Culture media therefor
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- A—HUMAN NECESSITIES
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- A23K20/111—Aromatic compounds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3481—Organic compounds containing oxygen
- A23L3/3508—Organic compounds containing oxygen containing carboxyl groups
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/42—Hydroxy-carboxylic acids
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- 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
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/66—Aspergillus
- C12R2001/685—Aspergillus niger
Abstract
The present invention relates to the manufacturing methods of new aspergillus niger A-T1 bacterial strain and the natural antimicrobial substance with this, specifically, have effects that new aspergillus niger (Aspergillus niger) A-T1 bacterial strain (deposit number KCCM 12147P) that plant-derived flavonoids (Flavonoids) is biologically converted into antibiotic property phenolic acid class (Phenolic acids) compound after hydrolysis) and, natural antimicrobial substance is manufactured using the A-T1 bacterial strain, preferably, generate the Phloroglucinol carboxylic acid (Phloroglucinol carboxylic acid) for manufacturing phenolic acid series and protocatechuic acid (Protocatechuic acid) and gallic acid (Gallic a Cid method).
Description
Technical field
The present invention relates to new aspergillus niger (Aspergillus niger) A-T1 bacterial strain and with this natural antimicrobial substance
Manufacturing method specifically has plant-derived flavonoids (Flavonoids) being biologically converted into antibiotic property phenol by hydrolysis
New aspergillus niger (Aspergillus niger) A-T1 bacterial strain of the effect of acids (Phenolic acids) compound (compile by deposit
Number KCCM 12147P)) and, utilize the A-T1 bacterial strain to manufacture natural antimicrobial substance, it is preferable that generate manufacture phenolic acid series
Phloroglucinol carboxylic acid (Phloroglucinol carboxylic acid) and protocatechuic acid (Protocatechuic acid)
With the method for gallic acid (Gallic acid).
Background technique
Influence of the antibiotic to treatment human diseases is very big, but asks at present because antibiotic residue and patience in human body occurs
The problems such as topic and the incurable disease that can not be treated with existing antibiotic, the natural antimicrobial substance aspect with novel mechanism
Research carry out relatively active.The natural antimicrobial substance is the secondary metabolites matter of traditional medicinal plant, including saponin(e,
Tannic acid, tannic acid, alkyl phenol, glycoalkaloid, flavonoids, sesquialter terpene lactone (Susquiterpenes lactones), terpene
(the Journal of Applied such as compound (Terpenoids), phorbol ester (Phorbol esters)
Pharmaceutical Science (applying pharmaceutical journal) 01 (06)；2011,16-20).
Wherein terpenoid (Flavonoids) is largely existed in the medicinal plants such as green tea, is had antibacterial, is resisted
Cancer, antiviral, antiallergy and anti-inflammatory isoreactivity, it was reported that substantially non-toxic.The terpenoid is by two phenyl
It is formed using the structure of pyranoid ring or three carbon atoms of structure similar with its in conjunction with medium, according to three carbon atoms
Structure is divided into flavones, flavonols, catechin etc..
The catechin (Catechins) is to belong in flavonoids flavan-3-alcohol (flavan-3- ol), antibacterium,
Antiviral and antimycotic and other effects prominent (Current Opinion in Biotechnology in September, 2012,23:174
~181, www.sciencedirect.com).The main compound of the catechin has epigallocatechin gallic acid
Ester (EGCG, 59%), epigallocatechin (EGC, 19%), L-Epicatechin gallate (ECG, 13.6%) and table
Theine (EC, 6.4%).
The compound of flavones series as described above passes through especially shown in catechin compounds for example following [chemical formula 1]
Biological hydrolysis is broken down into various phenolic acid compounds, further specifically, is broken down into Phloroglucinol carboxylic acid
(Phloroglucinol carboxylic acid) and protocatechuic acid (Protocatechuic acid) and gallic acid
(Gallic acid)。
Chemical formula 1
The Phloroglucinol carboxylic acid is also known as 2,4,6- trihydroxybenzoic acid (2,4,6- trihydroxybenzoic
Acid), efficacy against bacteria is prominent, according to report, to campylobacter jejuni (C.jejuni), colibacillus (E.coli), Dan Zeng
Listeria (L.monocytogens), enteron aisle detection of Salmonella (S.enterica) etc. have effects that powerful inhibition (Journal
Of Food Protection, Vol.66, No.10,2003, pp 1811~1821).
The protocatechuic acid is also known as 3,4-Dihydroxybenzoic acid (3,4-dihydroxybenzoic acid), from medicine
There is anti-inflammatory, anti-oxidant, anti-hypercalcinuria, antibacterium, antiviral, anticancer, anti-aging, anti-artery symptom, anti-swollen in Neo-Confucianism
Tumor, anti-asthma, antiulcer, anti-spasm and other effects.Wherein antibacterial actions mechanism is the decomposition function of bacterial membrane, antiviral work
It is that hepatitis B virus DNA is inhibited to synthesize and inhibit the discharge (Acta of HBsAg and HBeAg in HepG2 exquisiteness strain with mechanism
Poloniae Pharmaceutica-Drug Research (Polish Acta pharmacy-drug research), Vol. 72No.4pp-643
~650,2015).
The last gallic acid is also referred to as 3,4,5-trihydroxy benzoic acid (3,4,5- trihydroxybenzoic
Acid), exist in the form of phytochemicals (phytochemicals) in various plants, be applied not only to the receipts of human body
It holds back and stops blooding, also there is anti-nascent tumor and bacteriostatic activity, there is anti-melanin, anti-oxidant and anticancer activity (Molecules
2010,15,7985~8005).The gallic acid is in the concentration of 3~12ppm to methicillin-resistant staphylococcus grape
Coccus (MRSA) have antibiotic effect (American science communication, 2012；8(2), www.americanscience.org).
In addition described in existing KR published patent the 10-2014-0068351st (on 06 09th, 2014) by
Epigallo-catechin gallate (EGCG) is converted to the grass rhodotorula of the biotransformation capacity of L-Epicatechin gallate
(Rhodotorula graminis) JAG-12 [deposit number: KFCC11539P] and with this green-tea extract fermentation material
Manufacturing method.Utilize the JAG-12 bacterial strain, it is possible to produce bitter taste is reduced, the fermentation green tea that taste is promoted, further from table
Nutgall catechin gallic acid ester effectively produces L-Epicatechin gallate.
KR published patent the 10-2011-0096648th (on August 31st, 2011) describes one kind and will be deemed as micro- life
Training in the several bacterial strains of the fermentation strain of object fermented tea as the aspergillus niger of sociales (Aspergillus niger) bacterial strain
After nutrient solution is inoculated into green tea, the fermentation in Constant Temperature and Humidity Chambers (30 DEG C, 80%), and then have and conventionally manufactured fermented tea phase
As quality and shorten fermentation time microbial fermentation tea manufacturing method.
Prior art document
Patent document
(patent document 1) KR published patent the 10-2014-0068351st (on 06 09th, 2014)；
(patent document 2) KR published patent the 10-2011-0096648th (on August 31st, 2011).
Non-patent literature
(non-patent literature 1) Current Opinion in Biotechnology (the current commentary of biotechnology) 2012
Year September, 23:174~181.
(non-patent literature 2) Molecules (molecules) 2010,15,7985~8005.
(non-patent literature 3) Journal of Food Protection (food protection magazine), Vol. 66, No.10,
2003, pp 1811~1821.
(non-patent literature 4) Journal of American Science (American science communication), 2012；8(2).
Summary of the invention
Technical problem
Antibiotic property phenol is converted by biological hydrolysis by plant-derived flavonoids the purpose of the present invention is to provide a kind of
The novel microorganism bacterial strain of acids (Phenolic acids) compound.
Natural antibacterial is manufactured from plant-derived flavonoids using microorganism another object of the present invention is to provide a kind of
The method of substance.
Technical solution
The present invention, which provides to have, is biologically converted into antibacterial after hydrolysis for plant-derived flavonoids (Flavonoids)
The effect of property phenolic acid class (Phenolic acids) compound new aspergillus niger (Aspergillus niger) A-T1 bacterial strain.It is described
Aspergillus niger A-T1 bacterial strain (Aspergillus niger A-T1) arrives Zhong Jun association, (society) South Korea in deposit on November 06th, 2017
(KCCM), preservation address is the interior No. 2 street branch academic circle mansions of the great Ji in the western door zone in South Korea Seoul 45 buildings；Deposit number is KCCM
12147P。
The manufacturing method of present invention offer natural antimicrobial substance, characterized in that include: to utilize aspergillus niger
(Aspergillus niger) A-T1 bacterial strain (deposit number KCCM 12147P) is by plant-derived flavonoids
(Flavonoids) technique for obtaining antibiotic property phenolic acid class (Phenolic acids) compound by biological hydrolysis.
The present invention provides agriculture water livestock products compositions of additives, characterized in that includes: to utilize aspergillus niger
The natural antimicrobial substance of (Aspergillus niger) A-T1 bacterial strain (deposit number KCCM 12147P) production.
Beneficial effect
The beneficial effects of the present invention are,
Using aspergillus niger A-T1 bacterial strain of the invention, natural antimicrobial substance can be produced from plant-derived flavonoids, it is excellent
Selection of land produces antibiotic property phenolic acid compound, it is highly preferred that safely and effectively producing Phloroglucinol carboxylic acid and former catechu
Acid and gallic acid；
Phenolic acid compound made according to the present invention is natural antimicrobial substance, substantially non-toxic to body and environment, suppression
The growth of harmful bacteria processed and virus, antimycotic and anti-inflammatory effect is prominent, can be used as pesticide or animal feed, cosmetics, food
Product or drug material are widely applied.
Detailed description of the invention
Fig. 1 is the phylogenetic tree (phylogenic tree) of aspergillus niger A-T1 bacterial strain of the invention.
Specific embodiment
The present invention is described in detail below according to preferred embodiment.But following embodiment is only to illustrate structure and work of the invention
With being not to limit protection scope of the present invention.
Even implementing structure essential to the invention, but have been introduced in the prior art, or to technical field
Those of ordinary skill for obvious content be no longer to be described in detail.
Aspergillus niger A-T1 bacterial strain of the invention be from pick up from green tea cultivate separated in sample in regional corrosive ground after
It is cultivated on PDA agar medium, substrate mycelium (substrate mycelium) is white, aerial hyphae (aerial
It mycelium is) with the morphological feature for gradually becoming fulvescent by bottle green.
The aspergillus niger A-T1 is grown in the culture medium containing flavonoids, is had and is passed through the flavone compound
The effect of low molecule aromatic compound of phenolic acid series is converted to after biological hydrolysis.
The phenolic acid compound specifically includes Phloroglucinol carboxylic acid, protocatechuic acid, gallic acid, from pharmacologically having
There are strong antibacterial, antimycotic, antiviral and anti-inflammatory effect.
Therefore the antibiotic property phenolic acid compound manufactured using the aspergillus niger A-T1 bacterial strain by the flavonoids derived from plant
Various agriculture water livestock products compositions of additives, cosmetic composition, food or composite medicine etc. can be developed.
[embodiment]
(1) microbe to screen
In order to obtain the microorganism for decomposing flavonoids, using the corrosive ground for growing tea area of South Korea's Jizhou Island as sample
It obtains, this is diluted stage by stage in 0.9% sterile physiological saline solution.To microorganism separation culture medium be using
It is trained in YM (5.0g peptone, 3.0g yeast extract, 3.0g malt extract, 5.0g glucose, 15.0g agar, pH 6.5) agar
Support the agar medium for mixing in base and dividing after 1.0% flavonoids (catechin containing 60%) sterilizing.YM agar containing flavonoids
The sample is smeared on culture medium, under the conditions of 30 DEG C after stationary culture five days, by 105The micro- life of the characteristic of cfu/g or more
Object isolates and purifies in same culture medium.
(2) microorganism separates
In order to screen the excellent microorganism of flavonoids capacity of decomposition, the flavonoids agar medium containing indicator is manufactured
(20.0g catechin (60%), 15.0g agar, 0.01g bromophenol blue), is vaccinated with the purifying microorganism separated before this herein.It is yellow
After ketone decomposes, indicator, that is, bromophenol blue is turned yellow strongly by the acid ingredient of phenolic acid class, screens flavones using this phenomenon
The excellent microorganism of class capacity of decomposition, wherein one plant of separating mould, is ' A-T1 ' by the Strain Designation.
(3) 18S rRNA analysis and identification
Using Wizard genomic DNA purification kit (guide genomic DNA purification kit,
Promega, USA) the separation A-T1 bacterial strain chromosomal DNA (chromosomal DNA), using being used for 18S rDNA sequence
Universal primer (universal primer) i.e. NS1 (5'-GTAGTCATATGCTTGTCTC-3') and NS8 (5'- of column
TCCGCAGGTTCACC TACGGA- 3') primer amplification PCR.Then using Wizard SV Gel (guide SV gel) and
PCR clean-up system (PCR purification system, Promega, USA) refines the PCR product of amplification, utilizes ABI
PRISM3730XL DNA Analyzer (DNA analysis device) analyzes the PCR product sequence of refining.The A-T1 bacterial strain
18S rDNA be totally 1,645bp, be indexed in attachment sequence catalogue 1.
Using BLASTN program by the ribosomal DNA sequence (ribosomes of the sequencing results and GENEBANK
DNA sequence dna) it is compared.The similitude of sequence is using Clustal X and Mega 2program comparative analysis as a result, institute
Shown in the phylogenetic tree (phylogenic tree) for stating A-T1 bacterial strain such as attached drawing 1, it is accredited as aspergillus niger.
(reference: 1 thompson, J.D., John Higgins, D.G. and Ji Busen, T.J. (1994).Cruise tal fibre W:
improving the sensitivity of progressive multiple sequence alignment through
Sequence weighting (improves the sensitivity of progressive Multiple Sequence Alignment by sequence weighting), position-specific
Gap penalies and weight matrix choice (position specified difference away from reveal and weight matrix selection) .Nucleic
Acids Res. (nucleic acids research) 22,4673-4680.)
(4) Liquid Culture and solid culture of A-T1 bacterial strain
In order to analyze flavonoids and its catabolism substance, 2% flavonoids is put into YM broth bouillon (containing 60% youngster
Theine) mixture, after being inoculated with the A-T1 bacterial strain, liquid culture is obtained after cultivating four days under the conditions of 30 DEG C with 200rpm.
In addition, the moisture of camellia tree (Camellia) residue for extracting flavonoids (containing 15% catechin) is adjusted to
43%, it is inoculated with the A-T1 bacterial strain herein, after cultivating four days under the conditions of 25 DEG C, adds same amount of water, adds under the conditions of 50 DEG C
Pulverulent solids culture is obtained after heat drying.
(5) phenolic acid compound content analysis
A) analysis sample preparation
The liquid culture of the A-T1 bacterial strain takes supernatant liquor to be used as sample, and solid culture is mentioned with 70% alcohol
After taking and being concentrated under reduced pressure, the sample dissolved in the distilled water of same amount is used to analyze.Use contains in culture sample liquid or extracting solution
The distilled water (5 μ g/20 μ l) of 3,4- mesitylenic acid is diluted to 1ml.Diluted sample is put into the methyl hydroxylamine salt of 1mg
After hydrochlorate, reacted 30 minutes with 60 DEG C under alkaline condition.
Reaction mixture is acidified using 10% sulfuric acid solution of sodium chloride-containing as after 2 or less pH, with 4 μ l ether
It is extracted with 2 μ l ethyl acetates.It is put into the TEA of 15 μ l in extracting sample, after being concentrated with 40 DEG C using vaporized nitrogen, puts herein
Enter 20 μ l toluene, 20 μ l MTBSTFA, after mixed liquor is heated 30 minutes with 60 DEG C, prepares MO/ before GC-MS analysis
The sample of TBDMS form.
B) GC-MS is analyzed
Analysis instrument uses Agilent 6890N gas chromatographic analysis, and detector uses the selection inspection of Agilent 5975B mass
It surveys device (70eV, electron impact ion source).This outer analysis sample is analyzed with 50~650 μm of ranges with 0.99scans/s.
Analysis temperature is 260 DEG C of sample injector (injector), 300 DEG C of interface (interface), ion source (ion
Source) 230 DEG C, analytical column is to have used Ultra-2, cross-linked capillary column coated with
5%phenyl (cross-linked capillary column for being coated with 5% phenyl) -95%methyl polysiloxane boned phase
(95% methyl polysiloxane frame phase) (25m x 0.2 ㎜ I.D., 0.11 ㎜ film thickness Agilent
Technologies (0.11 ㎜ film thickness Agilent technology), Santa Clara, CA, USA).
Carrier gas uses helium, and flow (flow rate) is 0.5ml/min, uses the sample of 1 μ l.The temperature of baking oven is out
Begin to increase at 100~250 DEG C by 5 DEG C/min for two minutes, is then adjusted to 300 DEG C points by 20 DEG C/min within last five minutes
Analysis.
C) to the analysis result of Liquid Culture sample
[table 1]
To the Liquid Culture sample, from starting culture until by 90 hours, classification contains each phenolic acid according to the time period
Amount analyzed as a result, as shown above, with the passage of incubation time, 4-HBA (4-Hydroxybenzoic
Acid), Phloroglucinol carboxylic acid (Phlorog lucinol carboxylic acid) and protocatechuic acid (Protocatechuic
Acid it) is dramatically increased with the content of gallic acid (Gallic acid).
D) to the analysis result of solid culture sample
[table 2]
For the solid culture sample, after each personal 50% ethyl alcohol extracts before culture and after culture, with the liquid
The same Method Comparison of body culture sample as a result, shown in table 2 as above, including 4- hydroxybenzoic acid (4-
Hydroxybenzoic acids) the content of all kinds phenolic acid class (Phenolic acids) compound increase.
(6) antibacterial ability of amount of flavonoid metabolite matter is assessed
For the antibacterial ability for analyzing flavonoids and its metabolite, it is put into 2% flavonoids in YM broth bouillon and (contains
60% catechin), it is inoculated with after the A-T1 bacterial strain herein, cultivates four days under the conditions of 30 DEG C and 100~200rpm, then
Drying preparation analysis sample.
In addition, being inoculated with pathogenic bacteria i.e. colibacillus (KCCM 11234) and enteron aisle Salmonella in advance to the LB culture solution of 5ml
Bacterium (ATCC 53648) and vibrio parahemolyticus (KCCM 11965) are cultivated 17 hours under the conditions of 37 DEG C and 200rpm.
After adding the prepared analysis sample respectively by concentration in the LB culture solution of each 5ml, in 37 DEG C and 200rpm item
It is cultivated 16 hours under part, then measures absorbance (unit respectively under the conditions of 600nm；OD600nm), by the result be indexed to
Under in<table 3>.Control group is not add the analysis sample.
[table 3]
As above shown in<table 3>, analysis sample made according to the present invention inhibits the pathogenic bacteria with the increase of concentration
The increase of the effect of growth is significant.Therefore assert and provided by the invention hydroxyl is biologically converted by flavones by aspergillus niger A-T1
The low molecule aromatic compound of benzoic acid series all has excellent antibacterial effect.
Claims (4)
1. a kind of new aspergillus niger A-T1 bacterial strain, which is characterized in that
Flavonoids will be originated from by, which having effects that, is biologically converted into antibiotic property phenolic acid compound after hydrolysis.
2. a kind of manufacturing method of natural antimicrobial substance characterized by comprising
Plant-derived flavonoids is obtained into antibiotic property phenolic acid compound by biological hydrolysis using aspergillus niger A-T1 bacterial strain
Technique.
3. the manufacturing method of natural antimicrobial substance according to claim 2, which is characterized in that
The antibiotic property phenolic acid compound includes Phloroglucinol carboxylic acid, protocatechuic acid and gallic acid.
4. a kind of agriculture water livestock products compositions of additives characterized by comprising
The natural antimicrobial substance produced using aspergillus niger A-T1 bacterial strain.
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