CN106376597A - Multienzyme composite antibacterial agent - Google Patents
Multienzyme composite antibacterial agent Download PDFInfo
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- CN106376597A CN106376597A CN201610765402.XA CN201610765402A CN106376597A CN 106376597 A CN106376597 A CN 106376597A CN 201610765402 A CN201610765402 A CN 201610765402A CN 106376597 A CN106376597 A CN 106376597A
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- multienzyme
- compound preservative
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
Abstract
The invention relates to a multienzyme composite antibacterial agent. The multienzyme composite antibacterial agent mainly solves the problem that a common antibacterial agent in an external diagnostic kit cannot effectively inhibit mould, microzyme and actinomyces. The multienzyme composite antibacterial agent mainly comprises 0.1-20% of chitin endonuclease, 0.1-30% of beta-1, 3-glucanase, 0.01-10% of cellulase, 0.1-30% of phosphomannanase and 10-90% of EDTA (or EGTA). A bacterium-containing reagent is added into a solid composition or the bacterium-containing reagent is added into a solution so that the multienzyme composite antibacterial agent is obtained. The multienzyme composite antibacterial agent has effects of inhibiting various microbes and is sensitive to eukaryotic microbes. Compared with the antibiotic, the multienzyme composite antibacterial agent is environmentally friendly. The multienzyme composite antibacterial agent is degradable, does not produce residues, does not produce drug-resistant bacteria and does not produce toxicity and damage to chemical antiseptics such as sodium azide.
Description
Technical field
The present invention relates to a kind of multienzyme compound preservative, belong to multienzyme complex and suppress growth of microorganism field.
Background technology
Due to there is various albumen, nutrient substance in external diagnosis reagent case, so being easy to long bacterium, reagent is caused to lose
Effect, so various reagents all add antibacterial.
In test kit, conventional antibacterial has following a few class at present：
1) sodium azide, MVP, material, this kind of material is generally poisonous or even hypertoxic, and the target of suppression is mainly antibacterial, unwrapping wire
Bacterium.Inconspicuous to yeast, mycete bacteriostasis.
2) preservative such as sodium citrate, potassium sorbate, can substantially suppress the growth of yeast and mycete in acid condition,
But inhibitory action is relatively low in neutrality or alkaline environment.
3) various antibiotic are although most microorganism has corresponding antibiotic, but because antibiotic holds
It is also easy to produce fastbacteria, therefore it is not recommended that using.
In addition to a few class antibacterial above, someone is used lysozyme as antibacterial recently, and this antibacterial is only to gram
Positive bacteria has larger bacteriostasis, not good to gram negative bacteria inhibition, does not have substantially antibacterial effect to eukaryotic microorganisms
Really.
The compound enzyme antibacterial of the present invention mainly can suppress various growth of microorganism.To eukaryotic microorganisms especially yeast
Bacterium inhibition is more preferably.
Content of the invention
The technical problem to be solved in the present invention is to overcome the shortcomings of to commonly use antibacterial in mentioned reagent box, using variety classes
Enzyme different types of microorganisms is decomposed to cell wall and suppressed its growth.
The principle of institute of the present invention foundation：Saccharomycetic cell wall main component is glucosan, mannan and a small amount of fat
Class and protein, β -1,3 glucanase can decompose glucosan, and phosphomamlose carbohydrase can decompose mannan therein,
Make cell wall lysis eventually, so that cell is easily broken.The main component of fungal cell wall is chitin and cellulose, several in the present invention
Fourth matter restriction endonuclease and cellulase can decompose chitin in mycete (and partial yeast bacterium) cell wall respectively and cellulose makes
Cell is easily broken.Mg2+, Ca2+And some other trace element is the necessary prothetic group of microbial portion enzyme, it is also cell table
The ingredient in face, chelating agen EDTA or EGTA can make microorganism be difficult to grow and make cell with these ions of complexation after adding
Wall construction is loose to be more prone to rupture.
For achieving the above object, the multienzyme compound preservative main component of the present invention is：Chitin restriction endonuclease；β -1,3 Portugal
Dextranase；Cellulase；Phosphomamlose carbohydrase.Wherein, based on antibacterial gross weight, chitin restriction endonuclease 0.1%-20%；β-
1,3 glucanase 0.1%-30%；Cellulase 0.01%-10%；Phosphomamlose carbohydrase 0.1%-30%；Multienzyme is combined
It is also added with EDTA, EGTA, 10%-90% in antibacterial.
It can add reagent containing bacterium for solid-state composition, and during solid-state, additional proportion is 0.1-5g/L reagent.Can also be molten
Liquid status add reagent containing bacterium.
Brief description
Fig. 1 is different formulations of the present invention to various Antimicrobial rate figures
Fig. 2 is different formulations of the present invention to various Antimicrobial rate logarithmic charts
Specific embodiment
Embodiment 1：Multienzyme compound preservative formula 1 is to Pichia pastoris GS115, e. coli bl21, Staphylococcus aureus
Bacterium, Ma Nifei penicillium, the inhibitory action of streptomyces griseuses
Formula 1：
Weigh chitin restriction endonuclease 2mg (3.39%), β -1,3 glucanase 3mg (5.08%), cellulase 1mg respectively
(1.69%), phosphomamlose carbohydrase 3mg (5.08%), EDTA50mg (84.75%), gross weight 59mg.Add 100mM pH7.4
Kaliumphosphate buffer is dissolved to 10ml, final concentration of chitin restriction endonuclease 200mg/L, β -1,3 glucanase 300mg/L, fiber
Plain enzyme 100mg/L, phosphomamlose carbohydrase 300mg/L, EDTA 5000mg/L.This formula is formula 1.
(1) inhibitory action to Pichia pastoris GS115 for the multienzyme compound preservative formula 1
By multienzyme compound preservative (formula 1) 1ml, being added to 1ml bacteria concentration is 105CFU/ml Pichia pastoris GS115 bacterium
In liquid and 8mlYPD culture medium (antibacterial total amount is 0.59g/L), 30 degree of effect 16h, apply on YPD culture medium flat plate afterwards
Cloth, calculates bacteria concentration C1.It is 10 that matched group is added to 1ml bacteria concentration for 1ml water5CFU/ml Pichia pastoris GS115 bacterium solution and
In 8mlYPD culture medium, 30 degree of effect 16h, it is coated with YPD culture medium flat plate afterwards, calculate bacteria concentration C0.Bacteriostasis rate=C0/
C1.The final bacteriostasis rate of this experiment is 303.
(2) inhibitory action to e. coli bl21 for the multienzyme compound preservative formula 1
By multienzyme compound preservative (formula 1) 1ml, being added to 1ml bacteria concentration is 106CFU/ml e. coli bl21 bacterium solution
In 8ml LB culture medium, 37 degree of effect 5h, it is coated with LB solid medium flat board afterwards, calculate bacteria concentration C1.Matched group
Being added to 1ml bacteria concentration for 1ml water is 106In CFU/ml e. coli bl21 bacterium solution and 8ml LB culture medium, 37 degree of effect 5h,
It is coated with LB solid medium flat board afterwards, calculate bacteria concentration C0.Bacteriostasis rate=C0/C1.The final bacteriostasis rate of this experiment is 117.
(3) inhibitory action to staphylococcus aureuses for the multienzyme compound preservative formula 1
By multienzyme compound preservative (formula 1) 1ml, being added to 1ml bacteria concentration is 106CFU/ml staphylococcus aureuses bacterium
In liquid and 8ml nutrient broth medium (Carnis Bovis seu Bubali cream 3g water 1000mL peptone 5g PH 7.2～7.4), 37 degree of effect 5h, afterwards
Nutrient broth solid medium flat board is coated with, calculates bacteria concentration C1.Matched group is added to 1ml bacteria concentration for 1ml water
106In CFU/ml staphylococcus aureuses bacterium solution and 8ml nutrient broth medium, 37 degree of effect 5h, solid in nutrient broth afterwards
It is coated with body culture medium flat plate, calculate bacteria concentration C0.Bacteriostasis rate=C0/C1.The final bacteriostasis rate of this experiment is 128.
(4) inhibitory action to Ma Nifei penicillium for the multienzyme compound preservative formula 1
By multienzyme compound preservative (formula 1) 1ml, be added to 1ml OD600=0.1 Ma Nifei penicillium sp bacteria culture fluid and
8ml Czapek's medium (sodium nitrate 3g/L dipotassium hydrogen phosphate 1g/L magnesium sulfate (MgSO4 7H2O) 0.5g/L potassium chloride 0.5g/L sulfur
Acid ferrous iron 0.01g/L sucrose 30g/L), 25 degree of concussion 48h, absorbance (optical path 1cm) under measuring 600nm afterwards：OD1.Measure
Method is fermentation liquid 4000rpm centrifugation 5min, applicable PBS resuspended to 1ml, measure the absorbance under re-suspension liquid 600nm, divided by 10
It is fermentation liquid absorbance.Matched group is added to the Ma Nifei penicillium sp bacteria culture fluid of 1ml OD600=0.1 for 1ml water and 8ml examines
In family name's culture medium, 25 degree of concussion 48h, absorbance (optical path 1cm) under measuring 600nm afterwards：OD0.Bacteriostasis rate=OD0/OD1.OD
Assay method is ibid.The final bacteriostasis rate of this experiment is 267.
(5) inhibitory action to streptomyces griseuses for the multienzyme compound preservative formula 1
By multienzyme compound preservative (formula 1) 1ml, be added to 1ml OD600=0.1 streptomyces griseuses culture fluid and
8ml LA1 fluid medium (yeast extract 6.0%, glucose 4.0%, K2SO41.0%, MgSO41.0%, pH7.8,121 DEG C go out
Bacterium 30min), 30 concussion and cultivates 24h, measuring OD600 afterwards：OD1.OD assay method is with (4).Matched group adds for 1ml water
To in the streptomyces griseuses culture fluid and 8ml LA1 fluid medium of 1ml OD600=0.1,30 degree of concussion 24h, are surveying afterwards
Determine OD600：OD0.Bacteriostasis rate=OD0/OD1.OD assay method is with (4).The final suppression ratio of this experiment is 113.Embodiment 2：Multienzyme
Compound preservative formula 2 is to Pichia pastoris GS115, e. coli bl21, staphylococcus aureuses, Ma Nifei penicillium, Lycoperdon polymorphum Vitt
The inhibitory action of streptomycete
Formula 2：
Weigh chitin restriction endonuclease 10mg (17.18%), β -1,3 glucanase 8mg (13.74%), cellulase respectively
0.02mg (0.03%), phosphomamlose carbohydrase 0.2mg (0.34%), EDTA40mg (68.70%), gross weight 58.22mg.Add
100mM pH7.4 kaliumphosphate buffer is dissolved to 10ml, final concentration of chitin restriction endonuclease 1000mg/L, β -1,3 glucanase
800mg/L, cellulase 2mg/L, phosphomamlose carbohydrase 20mg/L, EDTA 4000mg/L.This formula is formula 2.
(1), to the inhibitory action method of Pichia pastoris GS115 with embodiment 1, this experiment is for multienzyme compound preservative formula 2
Whole bacteriostasis rate is 2314.
(2), to the inhibitory action method of e. coli bl21 with embodiment 1, this experiment is for multienzyme compound preservative formula 2
Whole bacteriostasis rate is 108.
(3) multienzyme compound preservative formula 2 to the inhibitory action method of staphylococcus aureuses with embodiment 1, this experiment
Final bacteriostasis rate is 105.
(4), to the inhibitory action method of Ma Nifei penicillium with embodiment 1, this experiment is for multienzyme compound preservative formula 2
Whole bacteriostasis rate is 89.
(5), to the inhibitory action method of streptomyces griseuses with embodiment 1, this experiment is final for multienzyme compound preservative formula 2
Bacteriostasis rate is 76.
Embodiment 3：Multienzyme compound preservative formula 3 is to Pichia pastoris GS115, e. coli bl21, Staphylococcus aureus
Bacterium, Ma Nifei penicillium, the inhibitory action of streptomyces griseuses
Formula 3：
Weigh chitin restriction endonuclease 0.1mg (0.17%), β -1,3 glucanase 0.5mg (0.83%), cellulase respectively
5mg (8.25%), phosphomamlose carbohydrase 15mg (24.75%), EDTA40mg (66.01%), gross weight 60.6mg.Add
100mM pH7.4 kaliumphosphate buffer is dissolved to 10ml, final concentration of chitin restriction endonuclease 10mg/L, β -1,3 glucanase
50mg/L, cellulase 500mg/L, phosphomamlose carbohydrase 1500mg/L, EDTA 4000mg/L.This formula is formula 3.
(1), to the inhibitory action method of Pichia pastoris GS115 with embodiment 1, this experiment is for multienzyme compound preservative formula 3
Whole bacteriostasis rate is 116.
(2), to the inhibitory action method of e. coli bl21 with embodiment 1, this experiment is for multienzyme compound preservative formula 3
Whole bacteriostasis rate is 99.
(3) multienzyme compound preservative formula 3 to the inhibitory action method of staphylococcus aureuses with embodiment 1, this experiment
Final bacteriostasis rate is 122.
(4), to the inhibitory action method of Ma Nifei penicillium with embodiment 1, this experiment is for multienzyme compound preservative formula 3
Whole bacteriostasis rate is 355.
(5), to the inhibitory action method of streptomyces griseuses with embodiment 1, this experiment is final for multienzyme compound preservative formula 3
Bacteriostasis rate is 72.
Embodiment 4：Reduce multienzyme compound preservative formula 1 consumption to Pichia pastoris GS115, e. coli bl21, golden yellow
Color staphylococcuses, Ma Nifei penicillium, the inhibitory action of streptomyces griseuses
Formula 1：With embodiment 1
(1) reduce the inhibitory action to Pichia pastoris GS115 for multienzyme compound preservative formula 1 consumption
By multienzyme compound preservative (formula 1) 0.2ml, being added to 1ml bacteria concentration is 105CFU/ml Pichia pastoris GS115
In bacterium solution and 8.8mlYPD culture medium (antibacterial total amount is 0.118g/L), 30 degree of effect 16h, afterwards in YPD culture medium flat plate
Upper coating, calculates bacteria concentration C1.It is 10 that matched group is added to 1ml bacteria concentration for 0.2ml water5CFU/ml Pichia pastoris GS115 bacterium solution
In 8.8mlYPD culture medium, 30 degree of effect 16h, it is coated with YPD culture medium flat plate afterwards, calculate bacteria concentration C0.Bacteriostasis rate
=C0/C1.The final bacteriostasis rate of this experiment is 260.
(2) reduce the inhibitory action to e. coli bl21 for multienzyme compound preservative formula 1 consumption
By multienzyme compound preservative (formula 1) 0.2ml, being added to 1ml bacteria concentration is 106CFU/ml e. coli bl21 bacterium
In liquid and 8.8ml LB culture medium, 37 degree of effect 5h, it is coated with LB solid medium flat board afterwards, calculate bacteria concentration C1.Right
Being added to 1ml bacteria concentration according to group for 0.2ml water is 106In CFU/ml e. coli bl21 bacterium solution and 8.8ml LB culture medium, 37
Degree acts on 5h, is coated with afterwards on LB solid medium flat board, calculates bacteria concentration C0.Bacteriostasis rate=C0/C1.This experiment is final to be pressed down
Bacterium rate is 60.
(3) reduce the inhibitory action to staphylococcus aureuses for multienzyme compound preservative formula 1 consumption
By multienzyme compound preservative (formula 1) 0.2ml, being added to 1ml bacteria concentration is 106CFU/ml staphylococcus aureuses
In bacterium solution and 8.8ml nutrient broth medium, 37 degree of effect 5h, it is coated with nutrient broth solid medium flat board afterwards, meter
Calculate bacteria concentration C1.It is 10 that matched group is added to 1ml bacteria concentration for 0.2ml water6CFU/ml staphylococcus aureuses bacterium solution and 8.8ml
In nutrient broth medium, 37 degree of effect 5h, it is coated with nutrient broth solid medium flat board afterwards, calculate bacteria concentration C0.
Bacteriostasis rate=C0/C1.The final bacteriostasis rate of this experiment is 42.
(4) reduce the inhibitory action to Ma Nifei penicillium for multienzyme compound preservative formula 1 consumption
By multienzyme compound preservative (formula 1) 0.2ml, it is added to the Ma Nifei penicillium sp bacteria culture fluid of 1ml OD600=0.1
With 8.8ml Czapek's medium, 25 degree of concussion 48h, absorbance (optical path 1cm) under measuring 600nm afterwards：OD1.Assay method is
With embodiment 1.Matched group is added to Ma Nifei penicillium sp bacteria culture fluid and the 8.8ml Cha Shi of 1ml OD600=0.1 for 0.2ml water
In culture medium, 25 degree of concussion 48h, absorbance (optical path 1cm) under measuring 600nm afterwards：OD0.Bacteriostasis rate=OD0/OD1.Experiment
Final bacteriostasis rate is 138.
(5) reduce the inhibitory action to streptomyces griseuses for multienzyme compound preservative formula 1 consumption
By multienzyme compound preservative (formula 1) 0.2ml, be added to 1ml OD600=0.1 streptomyces griseuses culture fluid and
8.8ml LA1 fluid medium (yeast extract 6.0%, glucose 4.0%, K2SO41.0%, MgSO41.0%, pH7.8,121 DEG C
Sterilizing 30min), 30 concussion and cultivates 24h, measuring OD600 afterwards：OD1.OD assay method is with embodiment 1.Matched group is
0.2ml water is added in the streptomyces griseuses culture fluid and 8.8ml LA1 fluid medium of 1ml OD600=0.1,30 degree of concussions
24h, is measuring OD600 afterwards：OD0.Bacteriostasis rate=OD0/OD1.The final suppression ratio of this experiment is 68.
Embodiment 5：Increase multienzyme compound preservative formula 1 consumption to Pichia pastoris GS115, e. coli bl21, golden yellow
Color staphylococcuses, Ma Nifei penicillium, the inhibitory action of streptomyces griseuses
Formula 1：With embodiment 1
(1) increase the inhibitory action to Pichia pastoris GS115 for multienzyme compound preservative formula 1 consumption
By multienzyme compound preservative (formula 1) 5ml, being added to 1ml bacteria concentration is 105CFU/ml Pichia pastoris GS115 bacterium
In liquid and 4ml 2 × YPD culture medium (antibacterial total amount is 2.95g/L), 30 degree of effect 16h, afterwards on YPD culture medium flat plate
Coating, calculates bacteria concentration C1.It is 10 that matched group is added to 1ml bacteria concentration for 5ml water5CFU/ml Pichia pastoris GS115 bacterium solution and
In 4ml 2 × YPD culture medium, 30 degree of effect 16h, it is coated with YPD culture medium flat plate afterwards, calculate bacteria concentration C0.Bacteriostasis rate
=C0/C1.The final bacteriostasis rate of this experiment is 4538.
(2) increase the inhibitory action to e. coli bl21 for multienzyme compound preservative formula 1 consumption
By multienzyme compound preservative (formula 1) 5ml, being added to 1ml bacteria concentration is 106CFU/ml e. coli bl21 bacterium solution
In 4ml 2 × LB culture medium, 37 degree of effect 5h, it is coated with LB solid medium flat board afterwards, calculate bacteria concentration C1.Comparison
Organizing and being added to 1ml bacteria concentration for 5ml water is 106In CFU/ml e. coli bl21 bacterium solution and 4ml 2 × LB culture medium, 37 degree of works
With 5h, it is coated with LB solid medium flat board afterwards, calculate bacteria concentration C0.Bacteriostasis rate=C0/C1.The final bacteriostasis rate of this experiment
For 266.
(3) increase the inhibitory action to staphylococcus aureuses for multienzyme compound preservative formula 1 consumption
By multienzyme compound preservative (formula 1) 5ml, being added to 1ml bacteria concentration is 106CFU/ml staphylococcus aureuses bacterium
In liquid and 2 times of concentration nutrient broth mediums of 4ml, 37 degree of effect 5h, apply on nutrient broth solid medium flat board afterwards
Cloth, calculates bacteria concentration C1.It is 10 that matched group is added to 1ml bacteria concentration for 5ml water6CFU/ml staphylococcus aureuses bacterium solution and 4ml
In 2 times of concentration nutrient broth mediums, 37 degree of effect 5h, it is coated with nutrient broth solid medium flat board afterwards, calculate bacterium
Concentration C0.Bacteriostasis rate=C0/C1.The final bacteriostasis rate of this experiment is 142.
(4) increase the inhibitory action to Ma Nifei penicillium for multienzyme compound preservative formula 1 consumption
By multienzyme compound preservative (formula 1) 5ml, be added to 1ml OD600=0.1 Ma Nifei penicillium sp bacteria culture fluid and
2 times of concentration Czapek's mediums of 4ml, 25 degree of concussion 48h, absorbance (optical path 1cm) under measuring 600nm afterwards：OD1.Mensure side
Method is same embodiment 1.Matched group is added to the Ma Nifei penicillium sp bacteria culture fluid of 1mlOD600=0.1 for 5ml water and 2 times of 4ml is dense
In degree Czapek's medium, 25 degree of concussion 48h, absorbance (optical path 1cm) under measuring 600nm afterwards：OD0.Bacteriostasis rate=OD0/
OD1.Testing final bacteriostasis rate is 592.
(5) increase the inhibitory action to streptomyces griseuses for multienzyme compound preservative formula 1 consumption
By multienzyme compound preservative (formula 1) 5ml, be added to 1ml OD600=0.1 streptomyces griseuses culture fluid and
4ml 2 times of concentration LA1 fluid mediums (yeast extract 6.0%, glucose 4.0%, K2SO41.0%, MgSO41.0%, pH7.8,
121 DEG C of sterilizing 30min), 30 concussion and cultivates 24h, measuring OD600 afterwards：OD1.OD assay method is with embodiment 1.Matched group
It is added in the streptomyces griseuses culture fluid and 2 times of concentration LA1 fluid mediums of 4ml of 1ml OD600=0.1 for 5ml water, 30
Degree concussion 24h, is measuring OD600 afterwards：OD0.Bacteriostasis rate=OD0/OD1.The final suppression ratio of this experiment is 378.
Specific embodiment described in the invention is only explanation for example to present invention spirit.The technical field of the invention
Technical staff can be made to described specific embodiment with various modifications or supplement or substituted using similar mode,
But the spirit without departing from the present invention or surmount scope defined in appended claims.
Table：Different formulations are to different microorganisms fungistatic effect table
Claims (9)
1. a kind of multienzyme compound preservative is it is characterised in that include following component：Chitin restriction endonuclease；Beta-1,3 glucanase；
Cellulase；Phosphomamlose carbohydrase.
2. multienzyme compound preservative as claimed in claim 1 it is characterised in that：Chitin restriction endonuclease is based on antibacterial gross weight
0.1%-20%.
3. multienzyme compound preservative as claimed in claim 1 it is characterised in that：Beta-1,3 glucanase is based on antibacterial gross weight
The 0.1%-30% of amount.
4. multienzyme compound preservative as claimed in claim 1 it is characterised in that：Cellulase is based on antibacterial gross weight
0.01%-10%.
5. multienzyme compound preservative as claimed in claim 1 it is characterised in that：Phosphomamlose carbohydrase is based on antibacterial gross weight
0.1%-30%.
6. multienzyme compound preservative as claimed in claim 1 it is characterised in that：Be also added with multienzyme compound preservative EDTA,
EGTA, its consumption is the 10%-90% based on antibacterial gross weight.
7. the multienzyme compound preservative described in a kind of any one of claim 1-6 purposes it is characterised in that：Multienzyme is combined suppression
Microbial inoculum is added to material containing fungus in solid form.
8. the multienzyme compound preservative described in a kind of any one of claim 1-6 purposes it is characterised in that：Multienzyme is combined suppression
Microbial inoculum is dissolved into solution form and is added to material containing fungus.
9. the multienzyme compound preservative described in a kind of any one of claim 1-6 purposes it is characterised in that：Multienzyme is combined suppression
Surface containing fungus is sprayed to after microbial inoculum dissolving.
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ID=57938461
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110250207A (en) * | 2019-06-19 | 2019-09-20 | 浙江万里学院 | A kind of citrus brown spot pathogenic bacteria bacteriostatic agent and preparation method thereof |
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| CN101040052A (en) * | 2004-09-10 | 2007-09-19 | 诺维信北美公司 | Methods for preventing, removing, reducing, or disrupting biofilm |
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Inventor after: Zhao Zhanyong Inventor after: Zheng Changlong Inventor before: Xu Zhanyong Inventor before: Zheng Changlong |
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Application publication date: 20170208 |