Qd nov. and application thereof

Abstract

The application discloses a new Amycolatopsis qudensis species and application. The Amycolatopsis qudensis D2301 provided by the application has a preservation number of CGMCC No. 26389 in the China General Microbiological Culture Collection Center, and has staphylococcus aureus inhibiting activity. The application has important significance for controlling gram-positive bacterial infection, safe production and storage of green agricultural products, maintaining a healthy ecological environment of crops, and protecting the health of animals and plants.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microorganisms, in particular to a new species of QD Amycolata and application thereof. BACKGROUND

[0002] Amycolatopsis is a large branch of the phylum Actinobacteria, belonging to the family Pseudonocardiaceae. Amycolatopsis was initially proposed by Lechevalier et al. in 1986 (LECHEVALIER MP, PRAUSER H, LABEDA 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.) and later revised by Lee and Tang et al. 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 2009; 59(6): 1401-1404; Tang S-K, Wang Y, Guan T-W, Lee J-C, Kim C-J, Li W-J. Amycolatopsis halophila sp. nov., a halophilic actinomycete isolated from a salt lake. International Journal of Systematic and Evolutionary Microbiology 2010; 60(5): 1073-1078.). Currently, there are 86 validly described species of Amycolatopsis, with Amycolatopsis orientates as the type species (https: / / lpsn.dsmz.de / genus / amycolatopsis).The basic taxonomic features of the genus Amycolatopsis are: gram-positive, non-acid-fast, mesophilic, aerobic; vegetative hyphae fragment into cuboidal elements; aerial hyphae present or absent, fragment into cuboidal elements or chains of spore-like elements, non-motile; no endospores and no mycelium; cell wall type IV, containing meso-DAP, whole-cell hydrolysate contains arabinose and galactose (sugar type A), no mycolic acids; phospholipid type II (containing PE and PME); major menaquinones MK-9 (H2, H4); (G+C) mol% of genomic DNA is 66-69%.

[0003] Amycolatopsis strains are a group of actinomycetes with important academic research value and commercial application value. It has been reported that Amycolatopsis strains can produce more than 100 kinds of bioactive compounds, some of which have been developed as drugs, and most of which have antibacterial and antitumor bioactivity (Berdy, J., Bioactive microbial metabolites. J Antibiot (Tokyo), 2005. 58(1): p. 1-26.). Vancomycin produced by Amycolatopsis is considered to be one of the strongest antibiotics against most gram-positive bacterial infections. Most of the Amycolatopsis strains are isolated from various soil habitats like Streptomyces (Saintpierre-Bonaccio, D., et al., Amycolatopsis plumensis 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 some species are isolated from clinical samples (Labeda, D.P., et al., Amycolatopsis kentuckyensis sp. nov. Amycolatopsis lexingtonensis sp. nov. and Amycolatopsis pretoriensis sp. nov, isolated from equine placentas. 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.(Goodfellow, M., et al., Amycolatopsis sacchari sp. nov., a moderately thermophilic actinomycete isolated from vegetable matter: Int J Syst Evol Microbiol, 2001. 51 (Pt 1): p. 187-93.), even ancient Roman catacombs (Groth, I., et al., Amycolatopsis nigrescens sp. nov., an actinomycete isolated from a Roman catacomb. Int J Syst Evol Microbiol, 2007. 57 (Pt 3): p. 513-9.). SUMMARY

[0004] The object of the present application is to provide a new Amycolatopsis sp. and application thereof.

[0005] In a first aspect, the present application claims a new Amycolatopsis sp.

[0006] The new Amycolatopsis sp. claimed in the present application is specifically Amycolatopsis qudensis D2301, which has a preservation number of CGMCC No. 26389 at China General Microbiological Culture Collection Center.

[0007] The Amycolatopsis qudensis D2301 is a gram-positive aerobic bacteria. On ISP2 medium, it develops rich substrate hyphae, which are often broken into short rods. The aerial hyphae are sparse, white, unbranched, broken into irregular rods, and differentiate into straight and short spore chains. The spores are short columnar and non-motile. No soluble pigment is produced. The strain D2301 has a growth tolerance range of 15-40℃, 0-7% NaCl, and pH 6.0-9.0, and the optimal growth conditions are 28℃, 0% NaCl, and pH 7.0.

[0008] In a second aspect, the present application claims a culture.

[0009] The culture claimed in the present application is a culture of the QD Amycolatopsis qudensis D2301 described in the first aspect above, which is a substance obtained by culturing the QD Amycolatopsis qudensis D2301 in an actinomycete culture medium.

[0010] In the culture described above, the substance includes the culture (the cell itself) of the QD Amycolatopsis qudensis D2301 and the metabolite of the culture of the QD Amycolatopsis qudensis D2301.

[0011] In the culture described above, the actinomycete culture medium can be a solid culture medium or a liquid culture medium.

[0012] The term "culture" refers to a general term for a liquid or solid culture medium in which a microbial population has been artificially inoculated and cultured. That is, a product obtained by growing and / or amplifying microorganisms, which can be a biologically pure culture of microorganisms, or can contain a certain amount of culture medium, metabolites, or other components produced during the culture process. The term "culture" also includes a subculture obtained by passing a microorganism, which can be a culture of a certain generation, or a mixture of several generations.

[0013] In the detailed description of the present application, the actinomycete culture medium is specifically an ISP2 culture medium.

[0014] In a third aspect, the present application claims a metabolite.

[0015] The metabolite claimed in the present application is a metabolite of the QD Amycolatopsis qudensis D2301 described in the first aspect above.

[0016] The term "metabolite" refers to primary metabolites and / or secondary metabolites produced during the metabolism of microorganisms. Primary metabolism refers to the process in which microorganisms absorb various nutrients from the outside world, and through catabolism and anabolism, generate substances and energy to maintain life activities. The product of primary metabolism is the primary metabolite, such as monosaccharides or monosaccharide derivatives, nucleotides, vitamins, amino acids, fatty acids, and various macromolecular polymers composed of them, such as proteins, nucleic acids, polysaccharides, and lipids. Secondary metabolism refers to the process in which microorganisms synthesize substances that have no clear function for life activities during a certain growth period, using primary metabolites as precursors. The product of secondary metabolism is the secondary metabolite, which is mostly a compound with a complex molecular structure. According to its function, it can be divided into antibiotics, hormones, alkaloids, and toxins.

[0017] In a fourth aspect, the present application claims a bacterial agent.

[0018] The bacterial agent claimed in the present application contains the QD Amycolatopsis qudensis D2301 described in the first aspect above, the culture described in the second aspect above and / or the metabolite described in the third aspect above.

[0019] In the above bacterial agent, the bacterial agent is a bacterial agent for inhibiting Staphylococcus aureus.

[0020] In the above bacterial agent, the bacterial agent contains a carrier in addition to the active ingredient. The carrier can be a carrier commonly used in the field of pesticides and biologically inert. The carrier can be a solid carrier or a liquid carrier; the solid carrier can be a mineral material, a plant material or a high molecular compound; the mineral material can be at least one of clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica and diatomite; the plant material can be at least one of corn meal, soybean meal and starch; the high molecular compound can be polyvinyl alcohol and / or polyglycol; the liquid carrier can be an organic solvent, a vegetable oil, a mineral oil or water; the organic solvent can be decane and / or dodecane.

[0021] In the above bacterial agent, the dosage form of the bacterial agent can be various dosage forms, such as liquid, emulsion, suspension, powder, granule, wettable powder or water dispersible granule.

[0022] If necessary, a surfactant (such as Tween 20, Tween 80, etc.), a binder, a stabilizer (such as an antioxidant), a pH adjuster, etc. can also be added to the bacterial agent.

[0023] In a fifth aspect, the present application claims the use of the QD Amycolatopsis qudensis D2301 described in the first aspect above, the culture described in the second aspect above or the metabolite described in the third aspect above or the bacterial agent described in the fourth aspect above in any one of the following:

[0024] (A1) inhibiting Staphylococcus aureus;

[0025] (A2) preparing a product for inhibiting Staphylococcus aureus;

[0026] (A3) preparing a medicament for treating and / or preventing a disease caused by Staphylococcus aureus infection;

[0027] (A4) against Gram-positive bacteria;

[0028] (A5) preparing an active product against Gram-positive bacteria;

[0029] (A6) preparing a medicament for treating and / or preventing a disease caused by a Gram-positive bacterial infection;

[0030] (A7) antibacterial;

[0031] (A8) preparing a product with antibacterial activity;

[0032] (A9) preparing a medicament for treating and / or preventing a disease caused by a bacterial infection;

[0033] (A10) safe production and / or storage of green agricultural products;

[0034] (A11) preparing a product for safe production and / or storage of green agricultural products;

[0035] (A12) maintaining a healthy ecological environment for crops;

[0036] (A13) preparing a product for maintaining a healthy ecological environment for crops;

[0037] (A14) protecting the health of animals and plants;

[0038] (A15) preparing a product for protecting the health of animals and plants.

[0039] In a sixth aspect, the present application claims a product for inhibiting Staphylococcus aureus.

[0040] The product for inhibiting Staphylococcus aureus claimed in the present application has an active ingredient of QD 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 inoculum as described in the fourth aspect above.

[0041] In a seventh aspect, the present application claims a product for anti-Gram-positive bacteria.

[0042] The product for anti-Gram-positive bacteria claimed in the present application has an active ingredient of QD 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 inoculum as described in the fourth aspect above.

[0043] In an eighth aspect, the present application claims a product for antibacterial.

[0044] The present invention claims a product for use against bacteria, the active ingredient of which is the QD Amycolatopsis qudensis D2301 of the first aspect above or the culture of the second aspect above or the metabolite of the third aspect above or the inoculum of the fourth aspect above.

[0045] In a ninth aspect, the present invention claims a method of inhibiting Staphylococcus aureus.

[0046] The method of inhibiting Staphylococcus aureus claimed by the present invention can comprise the step of treating a test sample with the QD Amycolatopsis qudensis D2301 of the first aspect above or the culture of the second aspect above or the metabolite of the third aspect above or the inoculum of the fourth aspect above.

[0047] The test sample is a sample containing Staphylococcus aureus.

[0048] The method is a non-diagnostic therapeutic method.

[0049] In a tenth aspect, the present invention claims a method of combating Gram-positive bacteria.

[0050] The method of combating Gram-positive bacteria claimed by the present invention can comprise the step of treating a test sample with the QD Amycolatopsis qudensis D2301 of the first aspect above or the culture of the second aspect above or the metabolite of the third aspect above or the inoculum of the fourth aspect above.

[0051] The test sample is a sample containing Gram-positive bacteria.

[0052] The method is a non-diagnostic therapeutic method.

[0053] In an eleventh aspect, the present invention claims a method of combating bacteria.

[0054] The method of combating bacteria claimed by the present invention can comprise the step of treating a test sample with the QD Amycolatopsis qudensis D2301 of the first aspect above or the culture of the second aspect above or the metabolite of the third aspect above or the inoculum of the fourth aspect above.

[0055] The test sample is a sample containing bacteria.

[0056] The method is a non-diagnostic therapeutic method.

[0057] In the detailed description of the present application, the Staphylococcus aureus is Staphylococcus aureus ATCC29213.

[0058] In the twelfth aspect, the present application claims the use of the QD Amycolatopsis D2301 in the first aspect above in the preparation of the culture in the second aspect above or the metabolite in the third aspect above or the microbial agent in the fourth aspect above.

[0059] Experiments have proved that the strain D2301 represents a new species of Amycolatopsis, named Amycolatopsis qudensis. The strain has Staphylococcus aureus inhibitory activity. The present application has important significance for controlling gram-positive bacterial infection, safe production and storage of green agricultural products, maintaining the healthy ecological environment of crops, and protecting the health of animals and plants.

[0060] Deposit Description

[0061] Classification and naming: Amycolatopsis qudensis;

[0062] Reference biological material: D2301;

[0063] Preservation agency: China General Microbiological Culture Collection Center;

[0064] Abbreviation of preservation agency: CGMCC;

[0065] Address: No. 3, Beichen West Road, Chaoyang District, Beijing;

[0066] Preservation date: January 6, 2023;

[0067] Preservation center registration number: CGMCC No. 26389. BRIEF DESCRIPTION OF DRAWINGS

[0068] Figure 1 Polar lipid TLC development map of strain D2301.

[0069] Figure 2 Phylogenetic tree constructed based on 16S rRNA gene sequences of strain D2301 and related strains.

[0070] Figure 3 Strain D2301 inhibits Staphylococcus aureus screening results. A is Escherichia coli fermentation broth; B is strain D2301 fermentation broth; C is Bacillus subtilis fermentation broth; D is vancomycin sensitive paper. DETAILED DESCRIPTION

[0071] The present application will be further described in conjunction with the specific embodiments, and the examples given are only to illustrate the present application, but not to limit the scope of the present application. The examples provided below can serve as a guide for further improvement by those of ordinary skill in the art, and do not in any way constitute a limitation on the present application.

[0072] The experimental methods in the following examples are all conventional methods, and are carried out according to the techniques or conditions described in the literature in the art or according to the product instructions, unless otherwise specified. The materials, reagents, etc. used in the following examples can be obtained commercially, unless otherwise specified.

[0073] Example 1, Isolation and screening and identification of strain D2301

[0074] I. Isolation of strain D2301

[0075] Strain isolation medium: sodium propionate 2 g / L, NH4NO3 0.1 g / L, KCl 0.1 g / L, MgSO4.7H2O 0.05 g / L, FeSO4.7H2O 0.05 g / L, marine trace salt 0.38 g / L, agar 15 g / L, pH 7.5.

[0076] Add amikacin (50 μg / L) and potassium dichromate (50 μg / L) as inhibitors. The growth of fungi and gram-negative bacteria in soil samples is inhibited.

[0077] The soil sample for strain isolation was collected from the rhizosphere of Panax notoginseng in Wenshan, Yunnan. The fresh soil sample was air-dried at room temperature for 2 weeks and then dried at 120°C for 15 min. 2 g of the dried soil was added to 18 mL of sterile normal saline and placed in a 28°C shaking bed for 40 min at a speed of 200 r / min to fully suspend the soil particles. Gradient dilution was performed to prepare 10 -4 Diluted soil sample suspension.

[0078] Strain isolation and purification: 0.2 mL was taken and spread on the isolation medium plate, which was incubated at 28°C for 4 weeks. After 4 weeks, different single colonies were picked according to the colony characteristics (shape, color, size, surface luster, etc.) to PYG medium plates (medium composition: peptone 3 g / L, yeast extract powder 5 g / L, glycerol 10 g / L, betaine 1.25 g / L, sodium pyruvate 1.25 g / L, agar 15 g / L, pH 7.5), and the culture was purified by the four-way streaking method. The obtained pure strain was preserved in liquid nitrogen and frozen at -80°C with 20% (v / v) glycerol as a protective agent.

[0079] In this experiment, strain D2301 was isolated and purified.

[0080] II. Observation of culture characteristics and detection of physiological and biochemical characteristics of strain D2301

[0081] Strain D2301 was cultured on modified ISP2 solid medium (formula: malt extract powder 10 g / L, yeast extract powder 4 g / L, glucose 4 g / L, calcium carbonate 2 g / L, agar 15 g / L, pH 7.5) at 28°C for 14 days, and the colony morphology and mycelial growth characteristics were observed on the 2nd, 4th, 7th, 10th, and 14th days, respectively. The growth temperature detection range was 4, 10, 15, 20, 25, 28, 30, 37, 40, 42, and 45°C; the growth salt concentration (NaCl) detection range was 0-10% and 15% (0-10 g / 100 mL and 15 g / 100 mL) in 12 concentration gradients (0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and 15); and the growth pH detection range was 4-10 in 7 gradients (4, 5, 6, 7, 8, 9, and 10) (Xu P, Li WJ, Tang SK, Zhang YQ, Chen GZ, et al. Naxibacter alkalitolerans 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 completed using the detection kits API 50CH and API ZYM produced by France Merier Company and the detection system GEN III produced by the United States BiOLOG Company. Other physiological characteristics of the strain, including gram staining properties, motility, oxygen demand, catalase activity, starch hydrolysis, gelatin liquefaction, indole production, H2S production, and cellulose hydrolysis activity, were mainly referred to Actinomycete Systematics: Principles, Methods and Practices (Xu L H. Actinomycete systematics: principles, methods and practices [M]. Beijing: Science Press, 2007, 93-108.).

[0082] The identification results show that the strain D2301 is a gram-positive aerobic bacteria. In the ISP2 medium, it develops rich intracellular hyphae, which is often broken into short rods. The aerial hyphae is sparse, white, unbranched, broken into irregular rods, and differentiates into straight and short spore chains. The spores are short columnar and non-motile. It does not produce soluble pigments. The growth tolerance range of the strain D2301 to temperature, NaCl and pH is 15-40℃, 0-7% NaCl, and pH 6.0-9.0, respectively, and the optimum growth conditions are 28℃, 0% NaCl, and pH 7.0. The strain D2301 is positive for catalase, esterase (C4), lipase (C8), lipase (C14), leucine arylamidase, valine arylamidase, acid phosphatase, and negative for oxidase, trypsin, cystine arylamidase, β-galactosidase, α-glucosidase, β-glucosidase, alkaline phosphatase, and naphthol-AS-BI-phosphohydrolase. It is positive for cellulose hydrolysis and starch hydrolysis, and negative for gelatin liquefaction, indole production reaction, and H2S production experiment. It can assimilate D-maltose, D-serine, D-fructose, D-mannose, glycerol, etc. as the sole carbon source and energy source.

[0083] III. Cell chemical characteristics detection of the strain D2301

[0084] The cell chemical components of strain D2301, such as cell wall amino acids, polar lipids, fatty acids and quinones were detected by TLC, GC and HPLC (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 chromatography 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 cell wall characteristic amino acid of strain D2301 was meso-DAP; the main polar lipid components were unknown phospholipids (PL), diphosphatidylglycerol (DPG), phosphatidylinositol (PI) and phosphatidylethanolamine (PE) Figure 1 ) and iso-C 16:0 (13.4%) and iso-C 16:0 2OH (29.8%), the detailed fatty acid composition of strain D2301 was shown in Table 1; the main respiratory quinone in the respiratory chain was MK-9(H4), and a small amount of MK-9(H2) was also detected.

[0085] Table 1. Cellular fatty acid composition of strain D2301 and its close relative Alkaliphilic Amycolata KCTC 49024T

[0086] Fatty acids (%) Strain D2301 Alcaligenes faecalis KCTC 49024 T ]]> 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: 0 2OH / 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

[0087] IV. Determination of the phylogenetic position of strain D2301

[0088] Genomic DNA of the strain D2301 was extracted for sequencing, and the 16S rRNA gene sequence (SEQ ID No. 1) thereof was subjected to online alignment in an international authoritative bacterial taxonomy analysis database (http: / / www.ezbiocloud.net / ) (Kim OS, 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 showed that the strain D2301 of the present application was most closely related to the species of the Amycolatopsis genus in the Amycolatopsidaceae family in the Actinobacteria phylum. Among them, the 16S rRNA gene sequence of the strain D2301 of the present application had the highest similarity of 95.6% to the Amycolapotsis alkalitolerans KCTC 49024 T in the database. This value was far below the limit of 98.65% for distinguishing prokaryotic microbial 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 Microbiol 2014; 64: 346-351.). The results preliminarily suggested that the strain D2301 of the present application might represent a new species. The 16S rRNA gene sequences of the related strains of the Amycolatopsidaceae family in the database were retrieved, and the strains were constructed into a phylogenetic tree by the MEGA software using the N-J method. The results showed that the strain D2301 fell within the Amycolatopsis genus evolutionary clade. On this basis, the 16S rRNA genes of the Amycolatopsis genus and the closely related strains of the strain D2301 were selected to construct a phylogenetic tree. The strain D2301 was closely related to the Amycolapotsis alkalitolerans KCTC 49024 T and Amycolapotsis pithecellobii RM 579 TStable clustering on one sub-branch, with Amycolapotsis alkalitolerans KCTC 49024 T the closest evolutionary distance Figure 2 ). Further, strain D2301 was compared with Amycolapotsis alkalitolerans KCTC 49024 T genome sequence, ANI and dDDH values were calculated. The ANI of strain D2301 with Amycolapotsis alkalitolerans KCTC 49024 T was 80.3%, and the dDDH was 23.4%. Both values were less than the defined values for distinguishing prokaryotic microbial species (ANI < 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 Microbiol 2014, 64, 346-351.), confirming that strain D2301 represented a new species of Amycolatopsis. The G+C content of strain D2301 was calculated from the genome sequence to be 69.5%. AntiSMASH analysis of the genome of strain D2301 found that D2301 contained rich biosynthetic gene clusters, with the potential to synthesize terpenes, polyketides, and glycopeptide compounds.

[0089] In summary, the culture characteristics, physiological and biochemical properties, cell chemical classification data (Table 1), 16S rRNA gene sequence information, and phylogenetic analysis of strain D2301 of the present application confirmed that it was a new species of Amycolatopsis, and its Latin name was named Amycolatopsis qudensis, and its Chinese name was QD Amycolatopsis. Strain D2301 is the type strain.

[0090] Amycolatopsis qudensis D2301 was deposited at the China General Microbiological Culture Collection Center on January 6, 2023, and its deposit number is CGMCC No. 26389.

[0091] IV. Antimicrobial activity determination of strain D2301

[0092] The antibacterial activity of the strain D2301 was detected by using Staphylococcus aureus subsp. aureus Rosenbach ATCC 29213 as the test bacteria. The antibacterial activity test was performed by using the paper disc diffusion method (Zhou DQ. Microbiology Experiment Manual [M]. Shanghai: Shanghai Science and Technology Press, 1986). The positive control was a 6 mm filter paper disc of vancomycin with a drug dose of 30 μg. The negative controls were Escherichia coli ATCC 25922 and Bacillus subtilis ATCC 6633, which were subjected to the same experimental conditions and procedures as the strain D2301.

[0093] The results of the determination showed that the strain D2301 had significant inhibitory activity against Staphylococcus aureus. Figure 3

[0094] The present application has been described in detail. For those skilled in the art, the present application can be implemented in a wider range under the same parameters, concentrations and conditions without departing from the spirit and scope of the present application and without performing unnecessary experiments. Although the present application provides specific examples, it should be understood that further improvements can be made to the present application. In summary, according to the principles of the present application, the present application is intended to include any changes, uses or improvements of the present application, including changes made by using conventional techniques known in the art, which are outside the scope disclosed in the present application. Some basic features can be applied within the scope of the following attached claims.​

Claims

1. Pseudomonas amylopectin ( Amycolatopsis qudensis D2301, characterized in that: The Amycolatopsis qudensis has a preservation number of CGMCC No. 26389 at China General Microbiological Culture Collection Center.

2. The fermentation broth of the Amycolatopsis qudensis of claim 1.

3. An inoculant characterized in that: The microbial agent contains the Amycolatopsis qudensis of claim 1 or the fermentation broth of claim 2.

4. The bacterial agent of claim 3, wherein: The microbial agent is a microbial agent for inhibiting Staphylococcus aureus.

5. The bacterial agent of claim 4, wherein: The Staphylococcus aureus is Staphylococcus aureus ATCC29213.

6. Use of the Amycolatopsis qudensis of claim 1 or the fermentation broth of claim 2 or the microbial agent of claim 3 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 a disease caused by Staphylococcus aureus infection; wherein, The use of (A1) is a non-disease-treating use.

7. Use according to claim 6, characterized in that: The Staphylococcus aureus is Staphylococcus aureus ATCC29213.

8. A product for inhibiting Staphylococcus aureus, whose active ingredient is the Amycolatopsis qudensis of claim 1 or the fermentation broth of claim 2 or the microbial agent of claim 3.

9. The product of claim 8, wherein: The Staphylococcus aureus is Staphylococcus aureus ATCC29213.

10. A non-disease-treating method for inhibiting Staphylococcus aureus, comprising the step of treating a test sample with the Amycolatopsis qudensis of claim 1 or the fermentation broth of claim 2 or the microbial agent of claim 3.

11. The method of claim 10, wherein: The Staphylococcus aureus is Staphylococcus aureus ATCC29213.

12. Use of the Amycolatopsis qudensis of claim 1 in preparing the fermentation broth of claim 2 or the microbial agent of claim 3.

Images