CN109221174B - Composition for synergistically inhibiting growth of bacteria and fungi - Google Patents
Composition for synergistically inhibiting growth of bacteria and fungi Download PDFInfo
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- CN109221174B CN109221174B CN201811152236.1A CN201811152236A CN109221174B CN 109221174 B CN109221174 B CN 109221174B CN 201811152236 A CN201811152236 A CN 201811152236A CN 109221174 B CN109221174 B CN 109221174B
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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
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/72—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms
- A01N43/80—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms five-membered rings with one nitrogen atom and either one oxygen atom or one sulfur atom in positions 1,2
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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
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
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Abstract
The invention discloses a composition for synergistically inhibiting the growth of bacteria and fungi. The composition for synergistically inhibiting the growth of bacteria and fungi comprises 1, 2-benzisothiazolin-3-one and toosendanin. Experimental research shows that the 1, 2-benzisothiazolin-3-one and the toosendanin are compounded to inhibit the growth of bacteria and fungi, the inhibition effect has better effect than that of any one component used alone, and the compound has obvious bacteriostasis and synergism in a certain range, can well improve the drug effect, reduce the using amount of the 1, 2-benzisothiazolin-3-one, and is beneficial to human health and environmental protection. The invention has simple formula, easy preparation, obvious effect and wide application range and prospect.
Description
The technical field is as follows:
the invention belongs to the field of chemistry and microorganisms, and particularly relates to a composition for synergistically inhibiting the growth of bacteria and fungi.
Background art:
various high-efficiency chemical bactericides such as quaternary ammonium salts, parabens (p-hydroxybenzoate esters), methylisothiazolinone and chlorinated kaempferon (kaempferon) thereof have good effect on inhibiting the growth of microorganisms, and in particular the isothiazolinone bactericides are widely added into the fields of cosmetics, washing products, personal care products and the like and have good antibacterial and bactericidal effects.
The preparation method of isothiazolinone bactericide is published by Goerdeler and Mittle in 1963, and developed and researched by Rohm and Haas company in 1972, and is first introduced into China and popularized and applied after 2015. Common isothiazolinone fungicides are: 1, 2-benzisothiazolin-3-one (BIT), 2-n-octyl-4-isothiazolin-3-One (OIT), 2-methyl-4-isothiazolin-3-one (MIT), 5-chloro-2-methyl-4-isothiazolin-3-one (CMIT). Isothiazolinone fungicides are chemically synthesized products of heterocyclic structures which inhibit or kill microorganisms by destroying DNA molecules within the microbial cell by virtue of active moieties on the heterocyclic ring. Isothiazolinone germicides are widely used in water treatment, cosmetics, paper making, light textile, water paint and other fields. The functions of the compounds are characterized by strong antibacterial ability, small application dosage, good compatibility, low toxicity and the like, and the compounds have strong antibacterial action on various bacteria and fungi; has the advantages of high efficiency, broad spectrum, low toxicity, good compatibility, wider pH application range, natural biodegradation and the like.
However, due to the extensive application and even abuse of the chemical bactericides, the drug resistance of pathogenic bacteria is becoming a common problem facing human beings, and at present, multi-drug resistant strains evolve to increase the probability that 'super bacteria' cannot be treated by available drugs. As fungal or bacterial skin diseases become increasingly serious and may endanger human life safety, human beings have been called to restrict the use of chemical bactericides on a large scale, new products have been actively developed, and new treatment methods have been sought to reduce or replace the use of chemical bactericides in daily use products. Therefore, innovative methods are imminent to develop new products for inhibiting bacterial or fungal infection, some special plants with inherent bactericidal capacity are concerned, and the potential trend of finding natural bactericides from plants for compounding and synergism is provided.
In searching for natural plantsIn the research on substances having bactericidal and insecticidal activities, a great deal of intensive research on neem has been conducted in the united states, india and other countries, and about 1600 or more plants have been found to have excellent bactericidal and insecticidal activities worldwide, among which the most studied are the plants of the family meliaceae, mainly neem, toosendan and neem. The toosendan fruit is a common green plant in Sichuan of China, and has the functions of dust retention, bacteriostasis and sterilization. The researchers of China began to research the chemical components, pharmacology, sterilization and disinsection effects and the like of the chinaberry (including the toosendan fruit) from the 50 th century. Toosendanin (Toosendanin) is a furantriterpenoid extracted from bark and fruit of Melia azedarach, and is composed of C, H, O, molecular weight is 574, and molecular formula is: c30H38O11The structure is shown as formula (I).
The toosendanin has high activity to fungi and agricultural pests as a natural bactericide and insecticide, and is easily decomposed in natural environment and is safe and harmless to human and livestock.
The invention content is as follows:
the invention aims to provide a composition for synergistically inhibiting the growth of bacteria and fungi. The novel compound is obtained by compounding the 1, 2-benzisothiazolin-3-one (BIT) and the natural plant bactericide, and the compound has the effect of improving the growth inhibition effect on bacteria and fungi, so that the use amount of the BIT is reduced.
The composition for synergistically inhibiting the growth of bacteria and fungi comprises 1, 2-benzisothiazolin-3-one and toosendanin.
The ratio of the amount of the 1, 2-benzisothiazolin-3-one to the amount of the toosendanin is preferably 1: 20-40.
The concentration of the 1, 2-benzisothiazolin-3-one is preferably 0.1-0.2 mM, and the concentration of the toosendanin is preferably 2-8 mM.
The bacteria in the composition for synergistically inhibiting the growth of bacteria and fungi are preferably bacteria of the genus Escherichia (Enterobacteriaceae), and the fungi are preferably fungi of the genus Aspergillus (Aspergillus) of the class Deuteromycetes (Sphaeropsidales).
The bacterium is preferably Escherichia coli (Escherichia coli)8099, and the fungus is preferably Aspergillus niger (Aspergillus niger) ATCC 16404.
The invention also provides application of the composition in preparing a medicament for inhibiting the growth of bacteria or fungi.
Preferably, the bacterium is a bacterium of the genus Escherichia (Escherichia) and the fungus is a fungus of the genus Aspergillus (Aspergillus).
The bacterium is Escherichia coli (Escherichia coli)8099, and the fungus is Aspergillus niger (Aspergillus niger) ATCC 16404.
Experimental research shows that the 1, 2-benzisothiazolin-3-one and the toosendanin are compounded to inhibit the growth of bacteria and fungi, the inhibition effect has better effect than that of any one component used alone, and the compound has obvious bacteriostasis and synergism in a certain range, can well improve the drug effect, reduce the using amount of the 1, 2-benzisothiazolin-3-one, and is beneficial to human health and environmental protection.
The invention has simple formula, easy preparation, obvious effect and wide application range and prospect.
The specific implementation mode is as follows:
the following examples are further illustrative of the present invention and are not intended to be limiting thereof.
Example test method
When the test strain in the following examples is Escherichia coli (Escherichia coli)8099, the test method is: the test strains were incubated for 24h in a 37 ℃ incubator with the normal NA slant medium (10 g of peptone, 5g of yeast powder, 10g of NaCl, 20g of agar powder, pH 7.0 per liter) of this example. Then washing the bacteria with sterile water, and diluting to OD600And (4) preparing a bacterial suspension working solution as 1.0. Pouring about 20mL of NA culture medium into a sterilized flat plate with the diameter of 90mm, horizontally standing and solidifying, inoculating 0.1mL of bacterial suspension working solution, and uniformly coating for later use.The test plate was punched with a sterilized punch having a diameter of 5.0mm, 50. mu.L of a sample to be tested was injected into the hole, incubated at 37 ℃ for 24 hours, and the size of the zone of inhibition was measured.
When the test strain in the following examples is Aspergillus niger ATCC 16404, the test method is: the test strains were cultured in a 30 ℃ incubator for 72 hours using PDA (Potato dextrose agar) slant medium in this example. Then washing the bacteria with sterile water, and diluting to OD600And (4) preparing a bacterial suspension working solution as 1.0. Pouring about 20mL of PDA culture medium into a sterilized flat plate with the diameter of 90mm, horizontally standing and solidifying, inoculating 0.1mL of bacterial suspension working solution, and uniformly coating for later use. The test plate was punched with a sterilized punch having a diameter of 5.0mm, 50. mu.L of a sample to be tested was injected into the hole, incubated at 30 ℃ for 72 hours, and the size of the zone of inhibition was measured.
Example 1:
a synergistic bacterial and fungal growth inhibiting composition comprises 0.1mM BIT and 2mM azadirachtin. The preparation method comprises the following steps: adding BIT and toosendanin into deionized water to make the final concentration of BIT 0.1mM and toosendanin 2mM, and mixing to obtain the composition for synergistically inhibiting the growth of bacteria and fungi.
Using Escherichia coli (Escherichia coli)8099 as a test strain, the test was conducted according to the test method described in the test method example above, and the measurement was made such that 0.2mM BIT solution (0.2mM BIT solution was prepared by adding BIT to deionized water to give a final concentration of 0.2mM BIT solution, and mixing to give 0.2mM BIT solution) had a bacteriostatic ring size against Escherichia coli (Escherichia coli)8099 of: 7.3 mm; the measured size of the inhibition cycle of a 4mM azadirachtin solution (4mM azadirachtin solution is prepared by adding azadirachtin to deionized water to a final concentration of 4mM, and mixing to obtain a 4mM azadirachtin solution) to Escherichia coli (Escherichia coli)8099 is as follows: 5.8 mm. The size of the inhibition ring of the composition for synergistically inhibiting the growth of bacteria and fungi to Escherichia coli (Escherichia coli)8099 in the embodiment is as follows: 8.5 mm.
It can be seen that the synergistic bacterial and fungal growth inhibiting composition of the present invention acts on Escherichia coli 8099, and has a better inhibiting effect than that of 0.2mM BIT solution or 4mM azadirachtin solution alone.
Example 2:
a synergistic bacterial and fungal growth inhibiting composition comprises 0.15mM BIT and 4mM azadirachtin. The preparation method comprises the following steps: adding BIT and toosendanin into deionized water to make the final concentration of BIT 0.15mM and toosendanin 4mM, and mixing to obtain the composition for synergistically inhibiting the growth of bacteria and fungi.
The measurement of the inhibition zone size of 0.3mM BIT solution (0.3mM BIT solution prepared by adding BIT to deionized water to give a final concentration of 0.3mM BIT solution and mixing to give 0.3mM BIT solution) to Escherichia coli (Escherichia coli)8099 was carried out by using Escherichia coli (Escherichia coli)8099 as a test strain according to the test method described in the test method example: 8.5 mm; the measured 8mM azadirachtin solution (8mM azadirachtin solution preparation method comprises adding azadirachtin into deionized water to make its final concentration be 8mM, and mixing to obtain 8mM azadirachtin solution), and the size of the inhibition ring of the solution on Escherichia coli (Escherichia coli)8099 is as follows: 7.0 mm. The size of the inhibition ring of the composition for synergistically inhibiting the growth of bacteria and fungi to Escherichia coli (Escherichia coli)8099 in the embodiment is as follows: 10.5 mm.
It can be seen that the synergistic bacterial and fungal growth inhibiting composition of the present invention acts on Escherichia coli 8099, and has a better inhibiting effect than that of 0.3mM BIT solution or 8mM azadirachtin solution alone.
Example 3:
a synergistic bacterial and fungal growth inhibiting composition comprises 0.2mM BIT and 8mM azadirachtin. The preparation method comprises the following steps: adding BIT and toosendanin into deionized water to make the final concentration of BIT 0.2mM and toosendanin 8mM, and mixing to obtain the composition for synergistically inhibiting the growth of bacteria and fungi.
Using Escherichia coli (Escherichia coli)8099 as a test strain, the test was conducted according to the test method described in the test method example above, and the measurement was made such that 0.4mM BIT solution (0.4mM BIT solution was prepared by adding BIT to deionized water to give a final concentration of 0.4mM BIT solution and mixing to give 0.4mM BIT solution) had a bacteriostatic ring size against Escherichia coli (Escherichia coli)8099 of: 11.3 mm; the measured size of the inhibition cycle of a 16mM azadirachtin solution (16mM azadirachtin solution is prepared by adding azadirachtin to deionized water to a final concentration of 16mM, and mixing to obtain a 16mM azadirachtin solution) to Escherichia coli (Escherichia coli)8099 is as follows: 7.6 mm. The size of the inhibition ring of the composition for synergistically inhibiting the growth of bacteria and fungi to Escherichia coli (Escherichia coli)8099 in the embodiment is as follows: 12.8 mm.
It can be seen that the synergistic bacterial and fungal growth inhibiting composition of the present invention acts on Escherichia coli 8099, and has a better inhibiting effect than that of the 0.4mM BIT solution or 16mM azadirachtin solution alone.
Example 4:
a synergistic bacterial and fungal growth inhibiting composition comprises 0.1mM BIT and 2mM azadirachtin. The preparation method comprises the following steps: adding BIT and toosendanin into deionized water to make the final concentration of BIT 0.1mM and toosendanin 2mM, and mixing to obtain the composition for synergistically inhibiting the growth of bacteria and fungi.
Aspergillus niger (Aspergillus niger) ATCC 16404 was used as the test strain and tested according to the test method of the test method example above, and the measured 0.2mM BIT solution (0.2mM BIT solution was prepared by adding BIT to deionized water to a final concentration of 0.2mM BIT solution and mixing to obtain 0.2mM BIT solution) had the following bacteriostatic ring size for Aspergillus niger (Aspergillus niger) ATCC 16404: 5.6 mm; the size of the bacteriostatic ring of 4mM azadirachtin solution (4mM azadirachtin solution) to Aspergillus niger (Aspergillus niger) ATCC 16404 was determined by adding azadirachtin to deionized water to a final concentration of 4mM and mixing to give a 4mM azadirachtin solution: 5.5 mm. The size of the inhibition loop of the synergistic bacterial and fungal growth inhibiting composition of this example to Aspergillus niger (Aspergillus niger) ATCC 16404 is: 7.0 mm.
It can be seen that Aspergillus niger ATCC 16404, which is treated with the synergistic bacterial and fungal growth inhibiting composition of the present invention, shows a better inhibiting effect than the 0.2mM BIT solution or the 4mM azadirachtin solution alone.
Example 5:
a synergistic bacterial and fungal growth inhibiting composition comprises 0.15mM BIT and 4mM azadirachtin. The preparation method comprises the following steps: adding BIT and toosendanin into deionized water to make the final concentration of BIT 0.15mM and toosendanin 4mM, and mixing to obtain the composition for synergistically inhibiting the growth of bacteria and fungi.
A test was performed according to the test method of the test method example above using Aspergillus niger (Aspergillus niger) ATCC 16404 as the test strain, and the bacteriostatic ring size of the 0.3mM BIT solution (0.3mM BIT solution was prepared by adding BIT to deionized water to a final concentration of 0.3mM BIT solution and mixing to give a 0.3mM BIT solution) against Aspergillus niger (Aspergillus niger) ATCC 16404 was determined as: 6.6 mm; the measured 8mM azadirachtin solution (8mM azadirachtin solution was prepared by adding azadirachtin to deionized water to a final concentration of 8mM, and mixing to give a 8mM azadirachtin solution) had a bacteriostatic ring size of Aspergillus niger (Aspergillus niger) ATCC 16404: 6.2 mm. The size of the inhibition loop of the synergistic bacterial and fungal growth inhibiting composition of this example to Aspergillus niger (Aspergillus niger) ATCC 16404 is: 7.6 mm.
It can be seen that Aspergillus niger ATCC 16404, which is treated with the synergistic bacterial and fungal growth inhibiting composition of the present invention, shows a better inhibiting effect than the 0.3mM BIT solution or the 8mM azadirachtin solution alone.
Example 6:
a synergistic bacterial and fungal growth inhibiting composition comprises 0.2mM BIT and 8mM azadirachtin. The preparation method comprises the following steps: adding BIT and toosendanin into deionized water to make the final concentration of BIT 0.2mM and toosendanin 8mM, and mixing to obtain the composition for synergistically inhibiting the growth of bacteria and fungi.
Aspergillus niger (Aspergillus niger) ATCC 16404 was used as the test strain and tested according to the test method of the test method example above, and the measured 0.4mM BIT solution (0.4mM BIT solution was prepared by adding BIT to deionized water to a final concentration of 0.4mM BIT and mixing to obtain 0.4mM BIT solution) had the following bacteriostatic ring size: 8.5 mm; the size of the bacteriostatic ring of a 16mM azadirachtin solution (16mM azadirachtin solution is prepared by adding azadirachtin to deionized water to a final concentration of 16mM, and mixing to obtain 16mM azadirachtin solution) to Aspergillus niger (Aspergillus niger) ATCC 16404 is as follows: 7.6 mm. The size of the inhibition loop of the synergistic bacterial and fungal growth inhibiting composition of this example to Aspergillus niger (Aspergillus niger) ATCC 16404 is: 10.0 mm.
It can be seen that Aspergillus niger ATCC 16404, which is treated with the synergistic bacterial and fungal growth inhibiting composition of the present invention, shows a better inhibiting effect than the 0.4mM BIT solution or the 16mM azadirachtin solution alone.
Claims (2)
1. The composition for synergistically inhibiting the growth of bacteria and fungi is characterized by comprising 1, 2-benzisothiazolin-3-one and toosendanin, wherein the concentration of the 1, 2-benzisothiazolin-3-one is 0.1-0.2 mM, and the concentration of the toosendanin is 2-8 mM; the bacterium is Escherichia coli (Escherichia coli)8099, and the fungus is Aspergillus niger (Aspergillus niger) ATCC 16404.
2. Use of a composition according to claim 1 in the manufacture of a medicament for inhibiting the growth of bacteria or fungi; the bacterium is Escherichia coli (Escherichia coli)8099, and the fungus is Aspergillus niger (Aspergillus niger) ATCC 16404.
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| CN106566443A (en) * | 2016-10-28 | 2017-04-19 | 天长市永泰密封材料有限公司 | Mouldproof and mothproof container waterborne sealant and preparing method thereof |
| CN107439617A (en) * | 2017-07-28 | 2017-12-08 | 张月婵 | A kind of bactericidal composition containing plant extracts |
| CN107468549A (en) * | 2017-10-13 | 2017-12-15 | 武汉百思凯瑞纳米科技有限公司 | A kind of bitter oily veepa oil nano-composition and its preparation method and application |
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