Preparation and application of a deep-sea streptomyces and its antifeedant and antifungal active substances

Abstract

The present application relates to the technical field of microbial pesticides, in particular to a deep-sea streptomyces with antifeedant function and antibacterial effect and application thereof. The streptomyces from the South China Sea deep sea is Streptomyces sp. NA13, which was preserved in China Center for Type Culture Collection on August 29, 2022, and the preservation number is CCTCC M 20221343. The strain has the ability to prepare antibiotics such as Antimycin, Candicidin, Naringenin, Surugamide, Fredericamycin, SAL-2242, Albaflavenone, Hopene, SGR PTMs, etc. In particular, the fermentation is used to prepare antimycin or cyclic octapeptide surugamide compounds, including novel antimycin compounds antimycin Q (1). The prepared antimycin compounds are used as insect pest antifeedants or antifungal drug lead compounds, and have application prospects of developing into natural source antibacterial and insecticidal pesticides.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of microbial pesticides, in particular to a deep-sea streptomyces with antifeedant function and antibacterial effect and application thereof. BACKGROUND

[0002] Actinomycetes are widely distributed in nature, and they can produce metabolites with various biological functions. It is worth noting that about 76% of the original molecular structure skeletons of commercially available antibiotics are derived from actinomycetes, among which the streptomyces genus is the most prominent (Journal of Industrial Microbiology Biotechnology, 2014, 41(2): 425-431). In 2019, Professor Li Dehai's team reported that streptomyces contributed 54% of the total number of new active secondary metabolites produced by actinomycetes (Current Medicinal Chemistry, 2020, 27(36): 6244-6273). More interestingly, Professor Zhu Weiming's team found that 67.3% of deep-sea streptomyces natural products showed biological activities such as cytotoxicity, antibacterial activity, and antimalarial activity (Oceanographic Collection, 2016, 51: 86-124).

[0003] Long-term and large-scale use of organic synthetic insecticides has led to the development of pesticide resistance in pests, causing serious environmental pollution and food safety problems (Science, 2002, 297:2222-2223). With the deepening of the concept of green control, people are also constantly exploring safer and more environmentally friendly pest control methods. The purpose of pest control is to effectively control the damage, and it is not the only means to kill pests. By using the method of antifeeding (or repellent), effective protection can also be achieved. Therefore, insect antifeeding agents have become an important alternative to organic insecticides. The cotton bollworm (Helicoverpa Armigera) is a worldwide pest that causes serious damage to cotton, African rice, corn and other food crops. Its larvae bore and eat the young growth points and reproductive organs of crops, and eat flower buds (Northern Gardening, 2020, 20:27-33). At present, chemical pesticides are mainly used to control cotton bollworm, but cotton bollworm has developed resistance to chemical pesticides, and chemical pesticides have slow degradation and residual effects. Therefore, the development and application of green and efficient, environmentally friendly natural antifeeding agents have attracted the attention of many scholars.

[0004] Plant fungal diseases have high incidence and wide range, and their occurrence causes the decline of agricultural product yield and quality, seriously affecting the development and safety of agricultural industry. For example, apple rot disease is a disease caused by the persistent weak host parasitic fungus Valsa ceratosperma, also known as canker disease (Modern Agricultural Science and Technology, 2008, 23: 147-147). The apple rot pathogen has the characteristics of latent infection, that is, when the fruit tree is healthy, the pathogen can remain in a latent state and is not easy to expand pathogenicity to the host; when the tissue around the infection point dies and the host vitality is weak, the pathogen can expand pathogenicity. This disease mainly occurs in mature orchards, causing apple yield decline and economic loss. At present, China has generally entered the high incidence period of apple rot disease, and about 80% of mature orchards have occurred rot disease (Wang Baojun, 2017, Master's Thesis of Anhui Agricultural University). Apple rot disease has become one of the important diseases on apple trees in China, which has long plagued fruit farmers, seriously restricted the yield of apples, caused serious economic losses, and has become an important disease limiting apple production and export in China.

[0005] At present, there are many commercial fungicides, and with the long-term use of traditional fungicides, plant pathogens have problems such as drug resistance and drug tolerance, and problems such as pesticide residues and environmental pollution need to be solved (Environmental Science & Technology, 2019, 53(7): 3347-3365). Therefore, developing fungicides with novel mechanisms of action, broad spectrum of action, and low resistance risk is the focus of fungicide development. SUMMARY

[0006] The present application provides a deep-sea streptomycete with antifeedant function and resistance to multiple pathogenic fungi and its application.

[0007] To achieve the above-mentioned purpose, the technical scheme adopted by the present application is:

[0008] A deep-sea streptomycete, Streptomyces sp. NA13 from the South China Sea, was deposited with the China Center for Type Culture Collection on August 29, 2022, and the deposit number is CCTCC M 20221343.

[0009] The deep-sea Streptomyces contains a gene cluster (7.10 Mb genome) for synthesizing antimycin, candicidin, naringenin, isorenieratene, indigoidine-like, ectoine, desferrioxamine B, surugamide A, fredericamycin A, SAL-2242, albaflavenone, hopene, SGR PTMs, diisonitrileantibiotic SF2768 skeleton compound.

[0010] Use of a deep-sea Streptomyces NA13 metabolite in the preparation of an antifeedant or an antifungal candidate drug.

[0011] Use of a deep-sea Streptomyces NA13 metabolite in the preparation of an antifeedant or an antifungal candidate drug.

[0012] Preparation of the deep-sea Streptomyces NA13 metabolite:

[0013] 1) The deep-sea Streptomyces NA13 is inoculated on ISP3 solid medium for activation for 2-5 days, then inoculated in ISP3 liquid medium for seed liquid at 28℃ for 2-4 days; then the fermentation seed liquid is inoculated in ISP3 liquid medium at 2-10% by volume percentage, and cultured at 28℃ for 5-10 days.

[0014] 2) The fermentation liquid is centrifuged to obtain the bacterial cells, and the active ingredients in the bacterial cells are extracted by butanone ultrasonic extraction for 30-50 min; after recovering the butanone solvent, crude extract A is obtained, which is the deep-sea Streptomyces NA13 metabolite.

[0015] The crude extract A is separated by rapid silica gel column chromatography, eluted with dichloromethane:methanol (v / v) in gradient of 100:0-0:100; the flow fraction F B After gel LH20 impurity removal, semi-preparative reverse phase HPLC is used, eluted with 80-90% methanol aqueous solution, and the flow rate is 2.0-4.0 mL / min, to obtain compounds antimycin Q (1), antimycin A1a (2), antimycin A2a (3), antimycin A3a (4), antimycin A4a (5), antimycin A7a (6); the flow fraction F CThe compound N-formylantimycic acid methyl ester (7) was obtained by semi-preparative reverse phase HPLC using gel LH20 as the impurity removal agent, eluted by 30-40% methanol aqueous solution at a flow rate of 2.0-3.0 mL / min; the eluate F F The compounds surugamide A (8), surugamide B (9), surugamide D (10), surugamide E (11) were obtained by semi-preparative reverse phase HPLC using gel LH20 as the impurity removal agent, eluted by 40-50% acetonitrile aqueous solution at a flow rate of 2.0-3.0 mL / min.

[0016] A new antimycin compound produced by a deep-sea Streptomyces, the antimycin compound is shown as (I),

[0017]

[0018] The application of an antimycin compound, the compounds antimycin Q (1), antimycin A1a (2), antimycin A2a (3), antimycin A3a (4), antimycin A4a (5), antimycin A7a (6) can be used as an antifeedant or an antifungal drug lead compound.

[0019] The fungus is

[0020] Valsa mali F68-1

[0021] Fusarium oxysporum f.sp.Cucurmerimum S19

[0022] The present application has the advantages of:

[0023] 1. The strain NA13 obtained by the present application has a unique nature, it comes from the deep-sea sediment of the South China Sea, and through whole genome sequencing and bioinformatics analysis, it is found that the genome contains gene clusters for synthesizing 25 kinds of compounds with antimycin skeleton, and has the potential to produce diverse active substances.

[0024] 2. The strain NA13 can produce antimycin or surugamide compounds at the same time; wherein the antifeedant function and the activity against plant pathogenic fungi of antimycin A1a (2), antimycin A2a (3), antimycin A3a (4), antimycin A4a (5), antimycin A7a (6) are invented for the first time.

[0025] 3. The antimycin Q is prepared by the metabolite of the strain NA13, and the compound is a new compound discovered for the first time, and the prepared antimycin substance can be used as a pest antifeedant or an antifungal drug lead compound, and has the application prospect of being developed into a natural source antibacterial and antiparasitic pesticide. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 The phylogenetic tree based on 16S rDNA sequence is provided for the application to show the taxonomic position of NA13.

[0027] Figure 2 The antimycin compounds secondary metabolites produced by the strain NA13 in the ISP3 fermentation medium are provided for the application.

[0028] Figure 3 The chemical structures of the compounds 1-11 obtained by the strain NA13 are provided for the application.

[0029] Figure 4 The HRESIMS spectrum of antimycin Q (1) obtained by the strain NA13 is provided for the application.

[0030] Figure 5 The 1H NMR, 13C NMR, HMQC, HMBC and NOESY of antimycin Q (1) obtained by the strain NA13 are provided for the application. 1 H- 1 HCOSY and HMBC are important correlations.

[0031] Figure 6 The experimental ECD spectrum and the calculated ECD spectrum of antimycin Q (1) obtained by the strain NA13 in CH3OH are provided for the application.

[0032] Figure 7 The antifeedant activity of antimycin compounds 1-6 on cotton bollworm. DETAILED DESCRIPTION

[0033] In order to better understand the content of the application, the following further describes in combination with specific embodiments, but the protection content of the patent is not limited to this.

[0034] Identification and characterization of Streptomyces abyssalis NA13

[0035] Strain NA13 was isolated from deep-sea sediment of South China Sea (E 113°2.353', N 13°58.498') and deposited in China Center for Type Culture Collection on August 29, 2022, with the accession number CCTCC M20221343. The strain grew vigorously on ISP3 and ISP4 media, and grew normally on ISP2 and Gause No.1 media. It formed round colonies and did not produce pigments (Table 1).

[0036] Table 1 Growth characteristics of Streptomyces abyssalis NA13

[0037] Medium Growth Aerial hyphae Submerged hyphae Soluble pigments ISP 2 Growth moderate Creamy yellow Flesh colour None ISP 3 Growth vigorous Pale yellow Flesh colour None ISP 4 Growth vigorous Creamy white Flesh colour None Gause No. 1 Growth moderate Pale yellow Flesh colour None

[0038] Streptomyces NA13 was inoculated into ISP3 liquid medium and cultured at 28°C for 36 h, and then total DNA was extracted according to the conventional method. Then, using the Whole Genome Shotgun (WGS) strategy, the Next Generation Sequencing (NGS) technology based on the Illumina NovaSeq sequencing platform, and the third-generation single-molecule sequencing technology based on the Oxford Nanopore ONT sequencing platform were used to obtain the complete genome map of strain NA13. The linear chromosomal genome size was about 6.91 Mb, the GC content was 73.44%, and it contained a linear plasmid with a size of about 186.13 kb and a GC content of 68.83%. BLAST analysis of its 16S rRNA gene sequence showed that it had a sequence homology of up to 100% with three standard strains of different species, S. daghestanicus NRRL B-5418 T , S. albidoflavus DSM 40455 T and S. violascens ISP 5183 T . Further phylogenetic tree constructed based on 16S rDNA sequence showed that these strains clustered in the same branch Figure 1 , indicating that they had the closest genetic evolutionary relationship. Finally, the Average Nucleotide Identity (ANI) value of the genome showed that the ANI of Streptomyces NA13 and S. albidoflavus DSM 40455 T reached 98.7%, indicating that they were of the same species. Therefore, strain NA13 was identified as Streptomyces albidoflavus.

[0039] wherein the 16S rRNA gene sequence 16S rDNA of Streptomyces albidoflavus NA13 is:

[0040] >Streptomyces albidoflavus NA13 16S rDNA

[0041]

[0042] Using antiSMASH analysis and manual curation it was found that strain NA13 harbors 25 secondary metabolite gene clusters with the potential to produce Antimycins, Candicidins, Naringenin, Isorenieratene, indigoidine-like, Ectoine, Desferrioxamine B, Surugamide A, Fredericamycin A, SAL-2242, Albaflavenone, Hopene, SGR PTMs, Diisonitrile antibiotic SF2768 backbone compounds, and the potential to synthesize various novel PKS, RiPP-like, NRPS, Terpene, and Siderophore-like compounds (Table 2). This indicates that NA13 has the ability to produce the types of compounds described in the table.

[0043] Table 2. The potential of strain NA13 to synthesize diverse secondary metabolites

[0044]

[0045] Example 2. Production of various types of bioactive substances by deep-sea Streptomyces NA13 and preparation methods

[0046] A series of antimycin and cyclic octapeptide-like secondary metabolites were obtained by fermentation of Streptomyces NA13 using ISP3 medium Figure 2 ); specifically:

[0047] Deep-sea Streptomyces NA13 was inoculated on ISP3 solid medium and cultured for 72 h for activation, then inoculated in ISP3 liquid medium and cultured at 28°C for 72 h as seed liquid; the fermentation seed liquid was inoculated in the same fermentation medium at a volume ratio of 1:18, and cultured at 28°C for 7 days with shaking. The fermentation broth was centrifuged to obtain the bacterial cells, and the active ingredients in the bacterial cells were extracted by butanone ultrasonic extraction for 30-50 min. After recovery of the butanone solvent, crude extract A was obtained, which was the metabolites of deep-sea Streptomyces NA13. Crude extract A was separated by flash silica gel column chromatography, eluted with dichloromethane:methanol (v / v) at a gradient of 100:0-0:100, and 7 fractions (F A -F G ) were obtained. Fraction F B was purified by gel LH20, and semi-preparative reverse phase HPLC was used to elute with 85% methanol aqueous solution at a flow rate of 3.0 mL / min, and compound antimycin Q (compound 1) was obtained (5.7 mg, t R:18.5min), antimycin A1a(2)(6.4mg,t R :20.2min), antimycin A2a(3)(8.0mg,t R :15.8min), antimycin A3a(4)(4.8mg,t R :12.5min), antimycin A4a(5)(5.3mg,t R :10.1min), antimycin A7a(6)(4.0mg,t R (11.5 min). The fraction F eluted at a ratio of 100:2 to 100:5 C After impurity removal using LH2O gel electrophoresis, the compound N-formylantimycic acid methylester(7) (2.8 mg, t) was obtained by semi-preparative reversed-phase HPLC followed by elution with 40% methanol aqueous solution at a flow rate of 2.5 mL / min. R (17.3 min). The eluted fraction F was 100:20. F After impurity removal using LH2O gel electrophoresis, semi-preparative reversed-phase HPLC was performed, followed by elution with 45% acetonitrile aqueous solution at a flow rate of 2.5 mL / min to obtain compound surugamide A(8) (52.3 mg, t). R :22.1min), surugamide B(9)(2.5mg,t R :16.5min), surugamide D(10)(3.8mg,t R :18.3min), surugamide E(11)(3.6mg,t R (19.4 min). The structure of the above compound is as follows: Figure 3 As shown.

[0048] The structures of the obtained compounds were analyzed. Specifically, compound 1 is a pale yellow powder, and HRESIMS data show a [M+H] structure with m / z 549.2809. + ion peak ( Figure 4 The corresponding molecular formula is C 28 H 40 N₂O₉ has an unsaturation degree of 10. The UV absorption at 228 nm and 317 nm in the UV spectrum is characteristic of antimycin compounds. Combining 1D NMR (DMSO-d₆) and HSQC spectra, the 28 carbons are assigned to 5 carbonyl carbon signals (δ¹⁸). C 179.4, 173.9, 169.7, 169.4 and 160.4), and 6 aromatic carbon signals (δ). C150.6, 126.9, 125.0, 123.1, 118.2 and 114.3), 7 methine carbon signals (δ C 75.3, 73.2, 72.0, 54.0, 49.7, 39.5 and 32.8), 4 methylene carbon signals (δ C 33.5, 28.6, 25.9 and 25.5), 6 methyl carbon signals (δ C 18.7, 17.8, 16.6, 15.2, 11.4 and 11.1), which indicated the presence of an antinomycin skeleton. NH-3'(δ H 9.81) and H-8'(δ H 8.31) HMBC correlation signals, the presence of an NH-CHO fragment could be deduced. 1 H- 1 H COSY correlation signals. According to a set of aromatic carbon hydrogen signals (δ C 114.3, δ C 150.6, δ C 126.9, δ H 8.23 / δ C 125.0, δ H 7.85 / δ C 123.1 and δ H 6.91 / δ C 118.2) (see Table 3) and NH-3'(δ H 9.81) and C-4'(δ C 125.0) HMBC correlation signals, the presence of an N-formyl-amino salicylic acid fragment could be deduced. 1 H- 1 H COSY correlation signals of H-7 / H-1 " / H-2" / H-3" / H-4" / H-5" / H-6" and HMBC correlation signals of H-6" with C-2" and C-4" in the HMBC spectrum, the presence of a 3-methyl hexane group ( Figure 5 ) could be deduced. 1 H- 1 H COSY correlation signals of H-2 " / H-3 " / H-4 " / H-5 " and HMBC correlation signals of H-8, H-3 " and H-5 " with C-1 " in the HMBC spectrum, the presence of a 2-methyl butyric acid group ( Figure 5 ) could be deduced. Since the compound has 10 degrees of unsaturation, removing 5 carbonyl signals and one benzene ring, the last degree of unsaturation should be a monocyclic ring. Combined with the presence of a 3-methyl hexane group ( 1 H- 1NH-3 / H-3 / H-4 / H-11 and H-10 / H-9 / H-8 / H-7 / H-1" in the H COSY spectrum and H-3, H-4 and H-9 with C-2, H-4 and H-7 with C-6 in the HMBC spectrum, it can be deduced that there is a nine-membered dilactone ring fragment Figure 5 ). Therefore, the planar structure of compound 1 is shown in Figure 5 . The relative configuration of compound 1 was determined according to the coupling constants and NOESY spectrum. The connections of H-3 / H-4, H-7 / H-9 / H-11, H-8 / H-10 were observed in the NOESY spectrum, and the large coupling constants (9-10 Hz) of H-7 / H-8 and H-8 / H-9 indicated that H-3, H-4, H-8 and H-10 were in one plane and H-7 and H-9 were in another plane Figure 5 ). The absolute configuration of the mother nucleus of compound 1 was 3S, 4R, 7R, 8R and 9S Figure 6 obtained by calculation of ECD. Therefore, compound 1 was named as antimycin Q as a new compound Figure 3 .

[0049] Table 3.1H NMR and 1 13C NMR data (600 MHz, DMSO-d6, δ in ppm, J in Hz) of compound 1 13

[0050] Position 1 H NMR ​ 13 C NMR ​ 2 169.4,C 3 5.32,t,(7.9) 54.0, CH 4 5.58,p,(6.5) 72.0, CH 6 173.9,C 7 2.54,m 49.7, CH 8 4.88,m 75.3, CH 9 4.93,m 73.2, CH 10 1.19,m 17.8, CH3 11 1.30,d.(6.7) 15.2, CH3 <!-- 6 --> 1′ 114.3,C 2′ 150.6,C 3′ 126.9,C 4′ 8.23,d,(7.9) 125.0, CH 5′ 6.91,t,(8.0) 118.3, CH 6′ 7.85,d,(8.1) 123.1, CH 7′ 169.7,C 8′ 8.31,d,(1.8) 160.4,C 1″ 1.46, m; 1.30, m 25.5, CH2 2″ 1.22,m 33.5, CH2 3″ 1.18,m 32.8, CH 4″ 1.18,m 28.6, CH2 5″ 0.78,m 11.1, CH3 6″ 0.78,m 18.7, CH3 1″′ 174.9,C 2″′ 2.45,m 39.5, CH 3″′ 1.63, m; 1.46, m 25.9, CH2 4″′ 0.86,t,(7.4) 11.4, CH3 5″′ 1.12,d,(7.2) 16.6, CH3 2'-OH 12.74, br s 3-NH 9.33, br s 3'-NH 9.81,s

[0051] Other known compounds antimycin A1a (2), antimycin A2a (3), antimycin A3a (4), antimycin A4a (5), antimycin A7a (6), N-formylantimycic acid methyl ester (7), surugamide A (8), surugamide B (9), surugamide D (10), surugamide E (11) Figure 3 were determined by 1D NMR spectrum and optical rotation data compared with the relevant literature.

[0052] Example 3 Anti-mycin compounds feeding deterrent activity assay

[0053] ​Determination method: Helicoverpa Armigera is selected to evaluate the antifeedant function. Determination method: First, punch the fresh and clean cabbage (Brassica chinensis) leaves with uniform thickness into 1cm diameter round leaf discs with a puncher. 50μL of the test compound solution (monomer compounds (compounds 1-6) three different concentrations are 1, 0.5, 0.25mg / ml respectively) is uniformly coated on the back of the leaf in the treatment group, and the same volume of methanol solution is coated in the blank control group. After the leaf discs are naturally dried, they are placed in a culture dish (diameter 9cm) pre-coated with filter paper for moisturizing. 2 pieces of treated leaf discs and 2 pieces of control leaf discs are placed in each culture dish, and they are placed in a cross pattern. 2 three-day-old Helicoverpa Armigera that have been starved for 12h are introduced into each culture dish, and the culture dish is placed in a 25℃ light incubator. The high concentration control is taken as the cutoff time when it is taken for 80% of the time. The area of each leaf disc taken for food is measured with a coordinate grid paper. The experiment is repeated at least 5 times or more. Calculate the antifeedant rate: antifeedant rate (%) = (AC-AT) / (AC+AT) x 100%, wherein AC and AT represent the areas of the leaf discs taken for food in the blank control group and the sample treatment group, respectively.

[0054] Experimental results: NA13 crude extract showed significant antifeedant activity (AI=83.33%) at 5mg / mL. Figure 7 As can be seen from Table 1, the antifungin compounds 1-6 mentioned in the present application have different degrees of antifeedant activity on Helicoverpa Armigera. In particular, compounds 1-4 have antifeedant activity (AI=87.83%, 88.46%, 89.86%, 90.79%) at 1mg / mL. It can be preliminarily inferred that the nine-membered double lactone ring structure of the antifungin parent nucleus is a necessary group for the activity.

[0055] Example 4 Determination of antifungal activity of antifungin compounds

[0056] Apple tree rot fungus Valsa mali F68-1 and Fusarium oxysporum f.sp. Cucurmerimum S19 are selected as the strains for evaluating the antifungal activity.

[0057] Determination method: the pathogenic fungi were inoculated to the center of PSA flat by block method, placed in a constant temperature incubator at 28°C, cultured for 2-3d, when the colony diameter was about 4cm, 10μL of test sample solution (256, 128, 64, 32, 16, 8, 4, 2, 1, 0.5 and 0.25μg / mL) was added to filter paper pieces placed symmetrically on the flat. The negative control was methanol solution, and the positive control was nystatin. The flat was placed in a constant temperature incubator at 28°C and cultured for 2d. The lowest test sample concentration of the sterile growth observed by naked eye was taken as the minimal inhibitory concentration (MIC), and each test was repeated three times.

[0058] Experimental results: the test results showed that the isolated antifungal compounds 1-6 all exhibited inhibitory activity to the two pathogenic fungi to different extents, but the activity was weak. Compound 7 had no inhibitory activity to the two pathogenic fungi (Table 4).

[0059] Table 4 Antifungal activity of compounds 1-7

[0060]

Claims

1. A deep-sea Streptomyces, characterized in that: The deep-sea Streptomyces is Streptomyces ( Streptomyces sp.)NA13, the Streptomyces was identified as Streptomyces microlepidocrocite (sp.)NA13. Streptomyces albidoflavus The novel coronavirus was deposited at the China Center for Type Culture Collection on August 29, 2022, with accession number CCTCC M 20221343.

2. The Streptomyces profundus according to claim 1, characterized in that: The deep-sea Streptomyces contains a gene cluster for synthesizing antimycin, candicidin, naringenin, isorenieratene, indigoidine-like, ectoine, desferrioxamine B, surugamide A, fredericamycin A, SAL-2242, albaflavenone, hopene, SGR PTMs, diisonitrile antibiotic SF2768 skeleton compound.

3. Use of the deep-sea Streptomyces of claim 1, characterized in that: Application of metabolites obtained by fermenting deep-sea Streptomyces NA13 and preparation of the metabolites into cotton bollworm antifeedants.

4. Use of the deep-sea Streptomyces of claim 1, characterized in that: Application of metabolites obtained by fermenting deep-sea Streptomyces NA13 and preparation of the metabolites into antimycin and surugamide, and the preparation method is as follows: 1) inoculate the deep-sea Streptomyces NA13 in claim 1 on ISP3 solid culture medium for activation for 2-5 days, then inoculate on ISP3 liquid culture medium, and culture at 28℃ for 2-4 days as seed liquid; inoculate the fermentation seed liquid into ISP3 liquid culture medium at 2-10% of volume percentage, and culture at 28℃ for 5-10 days under oscillation; 2) centrifuge the fermentation liquid to obtain bacterial cells, extract active ingredients in the bacterial cells by butanone ultrasonic for 30-50 min, and obtain crude extract A after recovering butanone solvent; The crude extract A was separated by flash silica gel column chromatography eluting with dichloromethane:methanol (100:0-0:100) gradient; fractions F v / v B purified by gel LH20 and semi-preparative reverse phase HPLC eluting with 80-90% methanol in water at a flow rate of 2.0-4.0 mL / min to obtain compounds antimycin Q (1), antimycin A1a (2), antimycin A2a (3), antimycin A3a (4), antimycin A4a (5), antimycin A7a (6); fractions F C purified by gel LH20 and semi-preparative reverse phase HPLC eluting with 30-40% methanol in water at a flow rate of 2.0-3.0 mL / min to obtain compounds N -formylantimycic acid methyl ester (7); fractions F F purified by gel LH20 and semi-preparative reverse phase HPLC eluting with 40-50% acetonitrile in water at a flow rate of 2.0-3.0 mL / min to obtain compounds surugamide A (8), surugamide B (9), surugamide D (10), surugamide E (11);​ The structural formula of compound antimycin Q (1) is ； The structural formula of compound antimycin A1a (2), antimycin A2a (3), antimycin A3a (4), antimycin A4a (5), antimycin A7a (6) is respectively ； Compound N The structural formula of the compound -formylantimycic acid methyl ester (7) is ； The structural formula of compound surugamide A (8), surugamide B (9), surugamide D (10), surugamide E (11) is respectively: 。

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