CN116286478A - Novel QD amycolatopsis species and application - Google Patents
Novel QD amycolatopsis species and application Download PDFInfo
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- CN116286478A CN116286478A CN202310070382.4A CN202310070382A CN116286478A CN 116286478 A CN116286478 A CN 116286478A CN 202310070382 A CN202310070382 A CN 202310070382A CN 116286478 A CN116286478 A CN 116286478A
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Abstract
The invention discloses a novel QD amycolatopsis species and application thereof. The preservation number of the QD amycolatopsis (Amycolatopsis qudensis) D2301 provided by the invention in the China general microbiological culture Collection center (CGMCC) is CGMCC No.26389. The strain has antibacterial activity of Staphylococcus aureus. The invention has important significance for controlling gram positive bacterial infection, safely producing and storing green agricultural products, maintaining healthy ecological environment of crops and protecting animal and plant health.
Description
Technical Field
The invention relates to the field of microorganisms, in particular to a novel QD amycolatopsis species and application thereof.
Background
Amycolatopsis (Amycolatopsis) is a large branch of the phylum actinomycetes, belonging to the Pseudonocardiaceae (Pseudomonas). Amycolatopsis was originally proposed by Lechevalier et al in 1986 to establish (LECHEVALIER MP, PRAUSER H, LABODAP DP, RUAN J-S.Two New Genera of Nocardioform Actinomycetes: amycolata gen. Nov. And Amycolatopsis gen. Nov. International Journal of Systematic and Evolutionary Microbiology 1986;36 (1): 29-37.), after which Lee and Tang et al have revised descriptions of the genus outlined in 2009 and 2010, respectively (Lee SD. Amycolatopsis ultiminotia sp. Nov., isolated from rhizosphere soil, and emended description of the genus Amycolatopsis. International Journal of Systematic and Evolutionary Microbiology; 59 (6): 1401-1404; tang S-K, wang Y, guan T-W, lee J-C, kim C-J, li W-J. Amycolatops halophilia. Nov., a halophilic actinomycete isolated from a salt lake. International Journal of Systematic and Evolutionary Microbiology; 60 (5): 1073-1078.). Currently, there are 86 species described effectively for Amycolatopsis species, with Amycolatopsis orientalis Amycolatopsis orientates as a model species (https:// lpsn. Dsmz. De/genus/amycolatopsis). The basic taxonomic features of amycolatopsis are: gram positive, not acid fast, moderate temperature, and aerobic; breaking the vegetative hyphae into tetragonal bodies; the aerial hyphae are or is not broken into tetragonal or spore-like chains, and do not swim; no endospores and mycelium bundles; cell wall type IV, meso-DAP containing, whole cell hydrolysate containing arabinose and galactose (saccharide type a), no mycolic acid; phospholipid type II (containing PE and PME): the major menaquinones are MK-9 (H2, H4): the (G+C) mol% content of the genomic DNA is 66 to 69%.
Amycolatopsis (Amycolatopsis) strains are a class of actinomycetes that have important academic research and commercial application values. Amycolatopsis strains have been reported to produce over a hundred bioactive compounds, some of which have been developed as pharmaceuticals, mostly with antibacterial, antitumor bioactivity (Berdy, J., bioactive microbial metatiles.J Antibiot (Tokyo), 2005.58 (1): p.1-26.). Vancomycin produced by amycolatopsis is considered one of the strongest antibiotics against most gram-positive bacterial infections. Most Amycolatopsis was isolated from a wide variety of soil habitats as Streptomyces (Saintpierre-Bonacccio, D., et al, amycolatopsis sp.nov., a novel bioactive actinomycete isolated from a New-Caledonian brown hypermagnesian ultramafic soil. Int J Syst Evol Microbiol,2005.55 (Pt 5): p.2057-61; kim, B., et al, amycolatopsis eurytherma sp.nov., a thermophilic actinomycete isolated from soil.int J Syst Evol Microbiol,2002.52 (Pt 3): p.889-94), and also parts of the seed strain were isolated from clinical samples (Labeda, D.P., et al, amycolatopsis kentuckyensis sp.Amycolatopsis lexemensis sp.nov.and Amycolatopsis pretoriensis sp.nov., isolatedframequinone places.int J Syst Evol Microbiol,2003.53 (Pt 5): p.1601-5; huang Y., et al, amycolatopsis palatopharyngis sp.nov., a potentially pathogenic actinomycete isolated from a human clinical source.int J Syst Evol Microbiol,2004.54 (Pt 2): p.359-63), plant tissue (Goodfe, M., et al, amycolatopsis sactric, a moderately thermophilic actinomycete isolated from vegetable matter: int J SystEvol Microbiol,2001.51 (1): p.187-93), and even Gregotia 3, et al, 54.97-93, et al, and so on.
Disclosure of Invention
The invention aims to provide a novel QD amycolatopsis species and application thereof.
In a first aspect, the invention claims a new species of amycolatopsis.
The new species of amycolatopsis claimed by the invention is specifically QD amycolatopsis (Amycolatopsis qudensis) D2301, and the preservation number of the novel species in the China general microbiological culture Collection center is CGMCC No.26389.
The QD amycolatopsis (Amycolatopsis qudensis) D2301 is a gram-positive aerobic bacteria. On ISP2 culture medium, the mycelia are developed to form abundant basal mycelia, the basal mycelia are often broken into short rods, aerial mycelia are sparse and white, the aerial mycelia are not branched and broken into irregular rods, the aerial mycelia are differentiated to form straight and short spore chains, and the spores are short, columnar and do not move. No soluble pigment is produced. Strain D2301 has tolerance ranges of 15-40deg.C, 0-7% NaCl, and pH 6.0-9.0 for temperature, naCl and pH, and the optimal growth conditions are 28deg.C, 0% NaCl, and pH7.0.
In a second aspect, the invention claims a culture.
The culture claimed in the present invention is the culture of the QD amycolatopsis (Amycolatopsis qudensis) D2301 described in the first aspect above, which is a substance obtained by culturing the QD amycolatopsis (Amycolatopsis qudensis) D2301 in actinomycete medium.
Among the above cultures, the substances include the culture of the QD amycolatopsis (Amycolatopsis qudensis) D2301 (cell itself) and the metabolite of the culture of the QD amycolatopsis (Amycolatopsis qudensis) D2301.
In the above culture, the actinomycete culture medium may be a solid medium or a liquid medium.
The term "culture" refers to a collective term for a liquid or solid medium in which a population of microorganisms has grown after artificial inoculation and cultivation. I.e. the product obtained by growing and/or amplifying the microorganism, which may be a biologically pure culture of the microorganism, or may contain a certain amount of medium, metabolites or other components produced during the culture. The term "culture" also includes subcultures obtained by passaging microorganisms, which may be a culture of a certain generation or a mixture of several generations.
In a specific embodiment of the invention, the actinomycete medium is specifically ISP2 medium.
In a third aspect, the invention claims a metabolite.
The claimed metabolites of the invention are those of QD amycolatopsis (Amycolatopsis qudensis) D2301 described in the first aspect above.
The term "metabolite" refers to a primary metabolite and/or a secondary metabolite produced during metabolism of a microorganism. Primary metabolism refers to a process in which microorganisms absorb various nutrients from the outside and produce substances and energy that maintain vital activities through catabolism and anabolism. The primary metabolite is primary metabolite such as monosaccharide or monosaccharide derivative, nucleotide, vitamin, amino acid, fatty acid, and various macromolecular polymers composed of the same, such as protein, nucleic acid, polysaccharide, and lipid. Secondary metabolism refers to the process of synthesizing substances which have no definite function on the life activities of microorganisms by taking primary metabolites as precursors in a certain growth period of microorganisms. The secondary metabolite is the secondary metabolite, and most of the secondary metabolites are compounds with relatively complex molecular structures. Depending on their action, they can be classified into antibiotics, hormones, alkaloids, toxins, etc.
In a fourth aspect, the invention claims a microbial agent.
The claimed microbial agents of the invention contain QD amycolatopsis (Amycolatopsis qudensis) D2301 as described in the first aspect above, a culture as described in the second aspect above and/or a metabolite as described in the third aspect above.
Wherein the microbial inoculum is a microbial inoculum for inhibiting staphylococcus aureus.
In the microbial inoculum, the microbial inoculum contains a carrier in addition to the active ingredient. The carrier may be a carrier commonly used in the pesticide arts and which is biologically inert. The carrier may be a solid carrier or a liquid carrier; the solid carrier can be mineral material, plant material or high molecular compound; the mineral material may be at least one of clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica, and diatomaceous earth; the plant material may be at least one of corn flour, soy flour and starch; the polymer compound may be polyvinyl alcohol and/or polyglycol; the liquid carrier may be an organic solvent, vegetable oil, mineral oil, or water; the organic solvent may be decane and/or dodecane.
Among the above-mentioned bacterial agents, the formulation of the bacterial agent can be various formulations, such as liquid, emulsion, suspending agent, powder, granule, wettable powder or water dispersible granule.
Surfactants (such as Tween 20, tween 80, etc.), binders, stabilizers (such as antioxidants), pH regulators, etc. can also be added into the microbial inoculum according to the need.
In a fifth aspect, the invention claims the use of QD amycolatopsis (Amycolatopsis qudensis) D2301 as described in the first aspect above or the culture as described in the second aspect above or the metabolite as described in the third aspect above or the microbial inoculum as described in the fourth aspect above in any one of the following:
(A1) Inhibiting staphylococcus aureus;
(A2) Preparing a product for inhibiting staphylococcus aureus;
(A3) Preparing a medicament for treating and/or preventing diseases caused by staphylococcus aureus infection;
(A4) Anti-gram positive bacteria;
(A5) Preparing an active product against gram-positive bacteria;
(A6) Preparing a medicament for the treatment and/or prophylaxis of diseases caused by gram-positive bacterial infections;
(A7) Antibacterial;
(A8) Preparing an antibacterial active product;
(A9) Preparing a medicament for the treatment and/or prevention of a disease caused by a bacterial infection;
(A10) Safe production and/or storage of green agricultural products;
(A11) Preparing a product for safe production and/or storage of green agricultural products;
(A12) Maintaining the healthy ecological environment of crops;
(A13) Preparing a product for maintaining a healthy ecological environment of crops;
(A14) Protecting animal and plant health;
(A15) The product for protecting animal and plant health is prepared.
In a sixth aspect, the invention claims a product for inhibiting staphylococcus aureus.
The product for inhibiting staphylococcus aureus claimed in the present invention has the active ingredient of QD amycolatopsis (Amycolatopsis qudensis) D2301 described in the first aspect, or the culture described in the second aspect, or the metabolite described in the third aspect, or the microbial inoculum described in the fourth aspect.
In a seventh aspect, the invention claims a product for combating gram positive bacteria.
The product for combating gram positive bacteria claimed in the present invention has as active ingredient QD amycolatopsis (Amycolatopsis qudensis) D2301 as described in the first aspect hereinbefore or a culture as described in the second aspect hereinbefore or a metabolite as described in the third aspect hereinbefore or a microbial inoculum as described in the fourth aspect hereinbefore.
In an eighth aspect, the invention claims a product for use in combating bacteria.
The product for antibacterial claimed in the present invention, whose active ingredient is QD amycolatopsis (Amycolatopsis qudensis) D2301 described in the first aspect above or the culture described in the second aspect above or the metabolite described in the third aspect above or the microbial inoculum described in the fourth aspect above.
In a ninth aspect, the invention claims a method of inhibiting staphylococcus aureus.
The method for inhibiting staphylococcus aureus claimed by the invention can comprise the following steps: the sample is treated with QD amycolatopsis (Amycolatopsis qudensis) D2301 as described in the first aspect above or the culture as described in the second aspect above or the metabolite as described in the third aspect above or the microbial inoculum as described in the fourth aspect above.
Wherein the sample to be tested is a sample containing staphylococcus aureus.
The method is a non-disease diagnostic therapeutic method.
In a tenth aspect, the invention claims a method of combating gram positive bacteria.
The method of combating gram positive bacteria as claimed in the present invention may comprise the steps of: the sample is treated with QD amycolatopsis (Amycolatopsis qudensis) D2301 as described in the first aspect above or the culture as described in the second aspect above or the metabolite as described in the third aspect above or the microbial inoculum as described in the fourth aspect above.
Wherein the test sample is a sample containing gram positive bacteria.
The method is a non-disease diagnostic therapeutic method.
In an eleventh aspect, the invention claims an antibacterial method.
The antibacterial method claimed in the present invention may comprise the steps of: the sample is treated with QD amycolatopsis (Amycolatopsis qudensis) D2301 as described in the first aspect above or the culture as described in the second aspect above or the metabolite as described in the third aspect above or the microbial inoculum as described in the fourth aspect above.
Wherein the sample to be tested is a sample containing bacteria.
The method is a non-disease diagnostic therapeutic method.
In a specific embodiment of the invention, the staphylococcus aureus is staphylococcus aureus ATCC29213.
In a twelfth aspect, the invention claims the use of QD amycolatopsis (Amycolatopsis qudensis) D2301 described in the first aspect above for the preparation of a culture described in the second aspect above or a metabolite described in the third aspect above or a bacterial agent described in the fourth aspect above.
Experiments have shown that the strain D2301 of the invention represents a new species of Amycolatopsis, designated QD Amycolatopsis (Amycolatopsis qudensis). The strain has antibacterial activity of Staphylococcus aureus. The invention has important significance for controlling gram positive bacterial infection, safely producing and storing green agricultural products, maintaining healthy ecological environment of crops and protecting animal and plant health.
Preservation description
Classification naming: amycolatopsis qudensis;
biological materials according to: d2301;
preservation mechanism: china general microbiological culture Collection center (China Committee for culture Collection);
the preservation organization is abbreviated as: CGMCC;
address: beijing, chaoyang area, north Chenxi Lu No.1, 3;
preservation date: 2023, 1, 6;
accession numbers of the preservation center: CGMCC No.26389.
Drawings
FIG. 1 shows a polar lipid TLC profile of strain D2301.
FIG. 2 is a phylogenetic tree constructed based on the 16S rRNA gene sequences of strain D2301 and related strains.
FIG. 3 shows the results of a screening for inhibition of Staphylococcus aureus by strain D2301. A is escherichia coli fermentation broth; b is the fermentation liquor of the strain D2301; c is bacillus subtilis fermentation broth; d is vancomycin drug sensitive paper.
Detailed Description
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Example 1 isolation screening and identification of Strain D2301
1. Isolation of Strain D2301
Strain isolation medium: sodium propionate 2g/L, NH 4 NO 3 0.1g/L，KCl 0.1g/L，MgSO 4 .7H 2 O 0.05g/L，FeSO 4 .7H 2 O0.05 g/L, marine trace salt 0.38g/L, agar 15g/L, pH7.5.
Aztreonam (50 μg/L) and potassium dichromate (50 μg/L) were added as inhibitors. Inhibit the growth of fungi and gram negative bacteria in soil samples.
Soil samples for strain separation are collected from the rhizosphere soil of pseudo-ginseng in Yunnan mountain.After the fresh soil sample was air-dried at room temperature for 2 weeks, it was dry-heated at 120℃for 15min. Taking 2g of dry and hot soil, adding into 18mL of sterile physiological saline, placing into a shaking table at 28 ℃ for 40min at a rotating speed of 200r/min to fully suspend soil particles, and performing gradient dilution to prepare 10 -4 Dilutions of soil sample suspension.
Separating and purifying strains: 0.2mL of the culture medium was spread on a plate of the isolation medium, and the culture was inverted at 28℃for 4 weeks. After 4 weeks, different single colonies were picked on PYG medium plates (medium composition: peptone 3g/L, yeast extract powder 5g/L, glycerol 10g/L, betaine 1.25g/L, sodium pyruvate 1.25g/L, agar 15g/L, pH 7.5) according to colony characteristics (shape, color, size, surface gloss, etc.), and the cultures were purified by the four-fold streaking method. The obtained pure strain is preserved in liquid nitrogen and frozen at-80 ℃ by taking 20% (v/v) glycerol as a protective agent.
In the experiment, the strain D2301 is obtained through separation and purification.
2. Culture characteristic observation and physiological and biochemical characteristic detection of strain D2301
Strain D2301 was cultured on modified ISP2 solid medium (formulation: malt extract 10g/L, yeast extract 4g/L, glucose 4g/L, calcium carbonate 2g/L, agar 15g/L, pH 7.5) at 28℃for 14 days, and the colony morphology and growth characteristics of hyphae were observed on days 2, 4, 7, 10, and 14, respectively. The growth temperature detection ranges are 4, 10, 15, 20, 25, 28, 30, 37, 40, 42 and 45 ℃; 12 (0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 and 15) concentration gradients ranging from 0-10% and 15% (0-10 g/100mL and 15g/100 mL) of growth salt concentration (NaCl); the growth pH detection range was 7 (4, 5, 6, 7, 8, 9, 10) gradients between 4 and 10 (Xu P, li WJ, tang SK, zhang YQ, chen GZ, et al Naxibacter alkilitolians gen. Nov., sp. Nov., a novel member of the family Oxalobacteraceae isolated from China. Int J Syst Evol Microbiol 2005; 55:1149-1153). The physiological and biochemical functions of the strain were performed using the detection kit API 50CH, API ZYM, manufactured by Meriera, france, and the detection system GEN III, manufactured by BiOLOG, USA. Other strain physiological characteristics including gram stain properties, motility, oxygen demand, thixotropic activity, starch hydrolysis, gelatin liquefaction, indole production、H 2 S production and cellulolytic Activity are mainly carried out by reference to the "actinomycetes System identification Manual" (Xu L H. Actinomyces systems: principles, methods and practices [ M)].Beijing:Science Press,2007,93-108.)。
The identification result shows that: strain D2301 is a gram-positive aerobic bacterium. On ISP2 culture medium, the mycelia are developed to form abundant basal mycelia, the basal mycelia are often broken into short rods, aerial mycelia are sparse and white, the aerial mycelia are not branched and broken into irregular rods, the aerial mycelia are differentiated to form straight and short spore chains, and the spores are short, columnar and do not move. No soluble pigment is produced. Strain D2301 has tolerance ranges of 15-40deg.C, 0-7% NaCl, and pH 6.0-9.0 for temperature, naCl and pH, and the optimal growth conditions are 28deg.C, 0% NaCl, and pH7.0. Positive for thixotropic enzyme, esterase (C4), lipoid esterase (C8), lipoid enzyme (C14), leucine arylamine enzyme, valine arylamine enzyme and acid phosphatase; oxidase, trypsin, cystine arylamidase, beta-galactosidase, alpha-glucosidase, beta-glucosidase, alkaline phosphatase, and naphthol-AS-BI-phosphate hydrolase; cellulose hydrolysis and starch hydrolysis are positive; gelatin liquefaction, indole production reaction and H 2 The S-production test was negative. Can assimilate D-maltose, D-serine, D-fructose, D-mannose, glycerol, etc. as the only carbon source and energy source.
3. Cytochemical characterization of strain D2301
Cell wall amino acids, polar lipids, fatty acids, and quinones of strain D2301 were detected by TLC thin layer chromatography, GC gas chromatography, HPLC liquid chromatography (Lechevalier, M.P).&Lechevalier,H.A.(1980).The chemotaxonomy of actinomycetes.In Actinomycete Taxonomy,pp.227–291.SIM Special Publication no.6.Edited by A.Dietz&D.W.Thayer.Fairfax,VA:Society for Industrial Microbiology；Sasser M.Identification of bacteria by gas ghromatography of cellular fatty acids,MIDI Technical Note 101.Newark,DE:MIDI inc；1990.Minnikin DE,O’Donnell AG,Goodfellow M,Alderson G,Athalye M et al.An integrated procedure for the extraction of bacterial isoprenoid quinones and polar lipids.J Microbiol Methods 1984; 2:233-241.). The amino acid characteristic of the cell wall of strain D2301 is meso-DAP; phospholipids (PL), biphospholipidyl glycerol (dpp), phosphatidylinositol (PI) and Phosphatidylethanolamine (PE) of unknown structure are the major polar lipid components (fig. 1); the dominant fatty acid is iso-C 16:0 (13.4%) and iso-C 16:0 2OH (29.8%), the fatty acid detailed composition of strain D2301 is given in table 1; the major respiratory quinone in the respiratory chain is MK-9 (H 4 ) A small amount of MK-9 (H) 2 )。
Table 1, strain D2301 and its related bacteria alkaline-resistant amycolatopsis KCTC 49024T cell fatty acid component
| Fatty acid (%) | Strain D2301 | Alkali- |
| iso-C 16:0 2OH | 29.8 | 19.1 |
| iso-C 16:0 | 13.4 | 39.5 |
| C 12:0 | 9.7 | 1.2 |
| C 16:1 w 7C | 7.8 | 4.1 |
| iso-C15:02OH/C 16:1 w 7C | 6.6 | 0.9 |
| C16:0 | 5.6 | 4.2 |
| iso-C 16:1 H | 5.0 | 0.8 |
| iso-C 15:0 | 4.7 | 2.8 |
| C 14:0 | 3.7 | 0.9 |
| anteiso-C 17:0 2OH | 2.3 | 1.9 |
| anteiso-C 17:0 | 2.0 | 9.0 |
| iso-C 14:0 | 2.0 | 6.8 |
| iso-C 17:0 | 1.3 | 0.8 |
4. Determination of the phylogenetic status of Strain D2301
Genomic DNA of strain D2301 was extracted and sequenced, and the 16S rRNA gene sequence (SEQ ID No. 1) therein was aligned on-line in the International authoritative bacterial taxonomic analysis database (http:// www.ezbiocloud.net /) (KimOS, cho YJ, lee K, et al 2012, introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species.Int J Syst Evol Microbiol,62:716-721.). The results show that: the present invention strain D2301 has a recent relationship with species of Amycolatopsis of the family Pseudonocardiaceae, the phylum actinomycetes. Wherein, the 16S rRNA gene sequence of the strain D2301 of the invention and alkali-resistant amycolatopsis Amycolapotsis alkalitolerans KCTC 49024 in a database T The highest similarity of (2) was 95.6%. This value is well below the limit of 98.65% for differentiating prokaryotic species (Kim M, oh HS, park SC, chun J. Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes.Int J Syst Evol Microbiol2014;64:346-351.). The results initially suggest that the strain D2301 of the present invention may represent a new species. And (3) taking the 16S rRNA gene sequence of the related strain of Pseudonocardiaceae in the database and the 16S rRNA gene sequence of the strain D2301, and constructing a phylogenetic tree for the strain by using MEGA software through an N-J method. The results show that strain D2301 falls within the amycolatopsis clade. On the basis, 16S rRNA genes of amycolatopsis and a strain D2301 related strain are selected, and a phylogenetic tree is constructed. Strain D2301 and alkali-resistant amycolatopsis Amycolapotsis alkalitolerans KCTC 49024 T Amycolapotsis pithecellobii RM 579 and 579 T Stably gather on a sub-branch and react with alkali-resistant amycolatopsis Amycolapotsis alkalitolerans KCTC 49024 T Is the closest to evolution (fig. 2). Further comparing the strain D2301 with the alkali-resistant amycolatopsis Amycolapotsis alkalitolerans KCTC 49024 T Genomic sequences, ANI and dDDH values were calculated. Strain D2301 and alkali-resistant amycolaapotsis alkalitolerans KCTC 49024 T ANI of (3) was 80.3% and dDDH was 23.4%. Both of these values are less than the defined value (ANI) for distinguishing between prokaryotic microorganism species<95-96％，dDDH<70%) (Kim, m.; oh, H.S.; park, s.c.; chun, J.Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes.Int J Syst Evol Microbiol2014,64,346-351.) confirms that strain D2301 represents a novel species of Amycolatopsis. The G+C content of strain D2301 was 69.5% calculated from the genome sequence. The genome of the strain D2301 is analyzed by anti-SMASH, and the D2301 is found to contain rich biosynthesis gene clusters and has the potential of synthesizing terpenes, polyketides and glycopeptides.
By combining the culture characteristics, physiological and biochemical characteristics, cytochemical classification data (Table 1) and 16S rRNA gene sequence information and systematic evolution analysis of the strain D2301, we confirm that the strain D2301 is a new species of amycolatopsis, named Latin Amycolatopsis qudensis and Chinese named QD amycolatopsis. Strain D2301 is a model strain.
Amycolatopsis qudensis D2301 is preserved in China general microbiological culture collection center (CGMCC) No.26389 in 2023, 1 and 6.
4. Determination of antibacterial Activity of Strain D2301
The antibacterial activity of strain D2301 was tested and the test strain was Staphylococcus aureus (Staphylococcus aureus subsp. Aureus Rosenbach) ATCC29213. The antibacterial activity was tested by the paper diffusion method (Zhou Deqing. Handbook of microbiology [ M ]. Shanghai: shanghai science and technology Press, 1986). Positive control: vancomycin drug sensitive experiment paper sheet is a 6mm filter paper sheet, and the drug dosage is as follows: vancomycin 30 μg. The negative controls were E.coli ATCC25922 and B.subtilis ATCC6633, both under the same experimental conditions and experimental procedures as those for strain D2301.
The assay results showed that strain D2301 had significant inhibitory activity against staphylococcus aureus (fig. 3).
The present invention is described in detail above. It will be apparent to those skilled in the art that the present invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with respect to specific embodiments, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.
Claims (10)
1. The amycolatopsis is characterized in that: the amycolatopsis is Amycolatopsis qudensis, the strain number is D2301, and the preservation number of the amycolatopsis in China general microbiological culture Collection center is CGMCC No.26389.
2. The culture of amycolatopsis according to claim 1, wherein the amycolatopsis according to claim 1 is cultured in an actinomycete medium.
3. A metabolite of amycolatopsis according to claim 1.
4. A microbial inoculum, characterized in that: the microbial inoculum comprises the amycolatopsis of claim 1, the culture of claim 2, and/or the metabolite of claim 3.
5. The microbial agent of claim 4, wherein: the microbial inoculum is used for inhibiting staphylococcus aureus.
6. Use of a amycolatopsis according to claim 1 or a culture according to claim 2 or a metabolite according to claim 3 or a bacterial agent according to claim 4 or 5 in any of the following:
(A1) Inhibiting staphylococcus aureus;
(A2) Preparing a product for inhibiting staphylococcus aureus;
(A3) Preparing a medicament for treating and/or preventing diseases caused by staphylococcus aureus infection;
(A4) Anti-gram positive bacteria;
(A5) Preparing an active product against gram-positive bacteria;
(A6) Preparing a medicament for the treatment and/or prophylaxis of diseases caused by gram-positive bacterial infections;
(A7) Antibacterial;
(A8) Preparing an antibacterial active product;
(A9) Preparing a medicament for the treatment and/or prevention of a disease caused by a bacterial infection;
(A10) Safe production and/or storage of green agricultural products;
(A11) Preparing a product for safe production and/or storage of green agricultural products;
(A12) Maintaining the healthy ecological environment of crops;
(A13) Preparing a product for maintaining a healthy ecological environment of crops;
(A14) Protecting animal and plant health;
(A15) The product for protecting animal and plant health is prepared.
7. A product for inhibiting staphylococcus aureus, the active ingredient of which is amycolatopsis according to claim 1 or the culture according to claim 2 or the metabolite according to claim 3 or the microbial inoculum according to claim 4 or 5;
or (b)
A product for combating gram positive bacteria, the active ingredient of which is a amycolatopsis according to claim 1 or a culture according to claim 2 or a metabolite according to claim 3 or a bacterial agent according to claim 4 or 5;
or (b)
A product for antibacterial use, comprising as an active ingredient the amycolatopsis of claim 1 or the culture of claim 2 or the metabolite of claim 3 or the microbial agent of claim 4 or 5.
8. A method of inhibiting staphylococcus aureus comprising the steps of: treating a sample with the amycolatopsis of claim 1 or the culture of claim 2 or the metabolite of claim 3 or the microbial inoculum of claim 4 or 5;
or (b)
A method of combating gram positive bacteria comprising the steps of: treating a sample with the amycolatopsis of claim 1 or the culture of claim 2 or the metabolite of claim 3 or the microbial inoculum of claim 4 or 5;
or (b)
An antibacterial method comprising the steps of: treatment of a sample with a amycolatopsis according to claim 1 or a culture according to claim 2 or a metabolite according to claim 3 or a bacterial agent according to claim 4 or 5.
9. The microbial agent of claim 5 or the use of claim 6 or the product of claim 7 or the method of claim 8, characterized in that: the staphylococcus aureus is staphylococcus aureus ATCC29213.
10. Use of amycolatopsis according to claim 1 for the preparation of a culture according to claim 2 or a metabolite according to claim 3 or a bacterial agent according to claim 4 or 5.
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