CN117050921B - Novel marine streptomycete for producing anthrax mycin and beta-erythromycin and application thereof - Google Patents

Novel marine streptomycete for producing anthrax mycin and beta-erythromycin and application thereof Download PDF

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CN117050921B
CN117050921B CN202311308003.7A CN202311308003A CN117050921B CN 117050921 B CN117050921 B CN 117050921B CN 202311308003 A CN202311308003 A CN 202311308003A CN 117050921 B CN117050921 B CN 117050921B
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张敬
张坤
丁文平
尹浩
凌娟
杨键
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Sanya Marine Ecological Environment Engineering Research Institute
South China Sea Institute of Oceanology of CAS
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South China Sea Institute of Oceanology of CAS
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Abstract

The invention discloses novel marine streptomycete for producing anthrax and beta-erythromycin and application thereof. The streptomyces is named as streptomyces [(s) ]Streptomycessp.) SCSIO K001 with deposit number: CGMCC No.27800. The invention separates a new streptomycete strain from Guangdong Zhujiang river sedimentStreptomycesSCSIO K001, the strain SCSIO K001 has the capability of producing anthracnose and beta-erythromycin, and the fermentation culture of the strain SCSIO K001 is utilized for separating and purifying to prepare the anthracnose and the beta-erythromycin, so that the method has the characteristics of low cost, simple process, environment friendliness, easiness in mass production and the like.

Description

Novel marine streptomycete for producing anthrax mycin and beta-erythromycin and application thereof
Technical Field
The invention belongs to the technical field of microorganisms, and in particular relates to novel marine streptomycete for producing anthrax mycin and beta-rubromycinStreptomycessp.) SCSIO K001 and uses thereof.
Background
In recent years, pathogen resistance has increased, and there is an urgent need to study and discover novel microorganisms that produce novel active secondary metabolites to combat the spread of new diseases and resistant pathogens. Due to the diversity of marine habitats and unique ecological factors, marine microorganisms are considered as excellent producers of a variety of biologically active secondary metabolites, providing important strain resources for the development of novel compounds with unique structures and strong biological activities.
Anthrax (CAS No. 1452849-94-3) is a novel compound from marine microorganisms discovered in recent years that can help researchers develop novel therapies for treating anthrax and other diseases, etc., such as methicillin-resistant Staphylococcus aureus-induced diseases, etc. The dichloro derivative produced after the chlorination of anthrax has good inhibitory activity on gram-positive bacteria (such as anthrax) and also has inhibitory activity on certain gram-negative bacteria. Anthrax provides a new backbone for the future development of new MRSA antibiotics. beta-Yuxithromycin (CAS No. 27267-70-5) is a bacterial metabolite isolated from Streptomyces, havingHas unique spiral ketone structure, has HIV-1 reverse transcriptase inhibiting activity, and has antibiotic activity to gram positive bacteria and other organisms, and is one kind of broad spectrum antibiotic. In addition, the human telomerase inhibitor is a high-efficiency human telomerase inhibitor, can specifically inhibit telomerase activity of cancer cells, and induce aging of the cancer cells so as to cause death of the cancer cells, thereby achieving the purpose of treating malignant tumors; at presentβ-erythromycin is considered as a promising precursor of anticancer drugs, with important research value.
Disclosure of Invention
The first object of the invention is to provide a novel marine streptomycete strain for producing anthrax and beta-Yu erythromycinStreptomycessp.) SCSIO K001 with deposit number: CGMCC No.27800. The streptomycete SCSIO K001 is separated from Guangdong Zhujiang river sediment in China, and the strain has the following taxonomic characteristics:
the colony is black brown on 2216E solid culture medium, and the mycelium in the medium is dark purple and the spore is powdery (shown in figure 1); the bacterial colony is dry, the single bacterial colony is generally round, is compact and difficult to diffuse, is convex in the middle, is compact in combination of the mycelium in the basal body and the aerial mycelium, is difficult to pick up, and is easy to scrape after the spore is produced at the end stage. On ISP2 culture medium, the colony is grey, the edge is slightly radial, the colony is compact, the middle is convex and thicker, the later colony is grey, the colony spreads to the periphery, the powder spores are powdery, and the mycelium in the basal body is purple; on ISP4 culture medium, the colony is white, the mycelium is thicker, the middle bulge is grey brown around mycelium in the medium, and the colony is grey and powdery spore in later period; aerial hyphae are gray on a Nutrient Agar culture medium, and surrounding hyphae in the medium are gray and powdery spores; on TSA medium, colonies were white, the edges of the colonies appeared off-white, the hyphae in the medium were yellow brown, and spores were produced less.
The strain SCSIO K001 grows under the conditions of 0-45 ℃ and pH 5.0-11.0 and 0-10% (w/v) NaCl. Tween 80, starch hydrolysis positive, while tween 20, 40, and 60, cellulose hydrolysis, milk clotting and liquefying, gelatin hydrolysis negative.
The antibacterial activity detection results of the 6 indicator bacteria show that the indicator bacteria have stronger antibacterial activity on staphylococcus aureus, bacillus subtilis and aspergillus niger, and have no obvious or no activity on pseudomonas aeruginosa, tubercle bacillus TB and candida albicans.
16S rRNA sequence similarity analysis using EzTaxon-e Server (www.ezbiocloud.net/identification), strain SCSIO K001 and StreptomycesStreptomyces collinusCGMCC 4.1623 T The similarity of (2) was 98.7%. Phylogenetic analysis also showed the strain SCSIO K001 and StreptomycesS. collinusCGMCC 4.1623 T The evolutionary distance is closest (as shown in fig. 2). In addition, the strain SCSIO K001 and the most similar speciesS. collinusCGMCC 4.1623 T The average nucleotide homology (ANI) and DNA-DNA hybridization (dDDH) values were 82.5% and 26.2%, respectively, well below the standard threshold for division of prokaryotic microorganism species (internationally recognized inter-species discrimination threshold ANI 95% -96%; dDDH 70%). It can thus be determined as a new species of Streptomyces.
A second object of the present invention is to provide Streptomyces [ ]Streptomycessp.) SCSIO K001 in the production of anthrax and beta-jade erythromycin.
The third object of the present invention is to provide a method of using StreptomycesStreptomycessp.) a method for producing anthracycline and beta-erythromycin by SCSIO K001, comprising the steps of: preparing a fermentation culture of streptomyces SCSIO K001; separating fermentation broth and mycelium of the fermentation culture, extracting the fermentation broth by butanone, and concentrating the butanone extract to obtain extract A; extracting mycelium with acetone, concentrating the acetone extract to obtain extract B; and (3) purifying the crude extract obtained by combining the extract A and the extract B through silica gel column chromatography and semi-preparative HPLC to obtain the compounds anthracnose mycin and beta-Yuerythromycin.
The fermentation culture is prepared by the following method: and inoculating streptomyces SCSIO K001 into a seed culture medium for culturing for 24-48 hours to obtain seed liquid, and inoculating the seed liquid into a fermentation culture medium for culturing for 144-240 hours to obtain a fermentation culture.
The optimal culture temperature of the strain is 28 ℃; the seed liquid is inoculated according to the volume ratio of 1:20.
The seeds areThe formula of the culture medium is as follows: 10 g/liter of soluble starch, K 2 HPO 4 1 g，MgSO 4 ·7H 2 O 1 g，(NH 4 ) 2 SO 4 2 g，CaCO 3 2 g of 10 times trace element solution 1 mL, peptone 1 g, yeast extract 0.5 g, sea salt 30 g, water as solvent and pH 7.2-7.4.
The formula of the 10 x trace element solution is as follows: 1.5 g/liter of ferrous oxide, 10 mL g/liter of 7.7M hydrochloric acid, 0.006 g g/liter of cobalt chloride, 0.036 g g/liter of manganese chloride, 0.024. 0.024 g g/liter of zinc chloride, 0.002. 0.002 g g/liter of boric acid, 0.19. 0.19 g g/liter of sodium molybdate, 0.1. 0.1 g g/liter of nickel chloride, 0.07 g/g g/liter of copper chloride, water as solvent and 6.0 pH.
The formula of the fermentation medium is as follows: yeast extract 4 g/L, malt extract 10 g/L, glucose 4 g/L, sea salt 30 g/L, and water as solvent.
A fourth object of the present invention is to provide StreptomycesStreptomycessp.) SCSIO K001 in the preparation of bacteriostat.
The bacteriostatic agent contains the streptomycete SCSIO K001 or a fermentation culture thereof as an active ingredient.
The invention has the following beneficial effects:
the invention separates a new streptomycete strain from Guangdong Zhujiang river sedimentStreptomycessp and SCSIO K001, the strain SCSIO K001 has the capability of producing anthracnose and beta-erythromycin, and the fermentation culture of the strain SCSIO K001 is utilized for separating and purifying to prepare the anthracnose and the beta-erythromycin, so that the method has the characteristics of low cost, simple process, environment friendliness, easiness in mass production and the like.
Streptomyces (Streptomyces sp.)Streptomycessp.) SCSIO K001 deposited at China general microbiological culture Collection center (CGMCC) at a position of 2023, 07, 05: the institute of microbiology, national institute of sciences, no. 3, north chen west way 1, region of korea, beijing city, postal code: 100101 and the preservation number is CGMCC No.27800.
Drawings
FIG. 1 shows StreptomycesStreptomycesMorphological features of sp, SCSIO K001.
FIG. 2 is a phylogenetic tree constructed based on the 16S rRNA gene sequence.
FIG. 3 shows the chemical structures of anthrax mycin (1) and beta-erythromycin (2).
Detailed Description
The following examples are further illustrative of the invention and are not intended to be limiting thereof.
Example 1: isolation and identification of strains
Separating and obtaining novel marine streptomycete for producing anthracycline and beta-erythromycin from Guangdong Zhujiang river sediment in ChinaStreptomycessp.) SCSIO K001. The taxonomic characteristics of streptomyces SCSIO K001 are:
the colony is black brown on 2216E solid culture medium, and the mycelium in the medium is dark purple and the spore is powdery (shown in figure 1); the bacterial colony is dry, the single bacterial colony is generally round, is compact and difficult to diffuse, is convex in the middle, is compact in combination of the mycelium in the basal body and the aerial mycelium, is difficult to pick up, and is easy to scrape after the spore is produced at the end stage. On ISP2 culture medium, the colony is grey, the edge is slightly radial, the colony is compact, the middle is convex and thicker, the later colony is grey, the colony spreads to the periphery, the powder spores are powdery, and the mycelium in the basal body is purple; on ISP4 culture medium, the colony is white, the mycelium is thicker, the middle bulge is grey brown around mycelium in the medium, and the colony is grey and powdery spore in later period; aerial hyphae are gray on a Nutrient Agar culture medium, and surrounding hyphae in the medium are gray and powdery spores; on TSA medium, colonies were white, the edges of the colonies appeared off-white, the hyphae in the medium were yellow brown, and spores were produced less.
The strain SCSIO K001 grows under the conditions of 0-45 ℃ and pH 5.0-11.0 and 0-10% (w/v) NaCl. Tween 80, starch hydrolysis positive, while tween 20, 40, and 60, cellulose hydrolysis, milk clotting and liquefying, gelatin hydrolysis negative.
The antibacterial activity detection results of the 6 indicator bacteria show that the indicator bacteria have stronger antibacterial activity on staphylococcus aureus (with a bacteriostasis circle diameter of >2 cm), bacillus subtilis (with a bacteriostasis circle diameter of >1 cm) and aspergillus niger (with a bacteriostasis circle diameter of >1.5 cm), and have insignificant or no activity on pseudomonas aeruginosa, tubercle bacillus TB and candida albicans (with a bacteriostasis circle diameter of <0.2 cm).
16S rRNA sequence similarity analysis using EzTaxon-e Server (www.ezbiocloud.net/identification), strain SCSIO K001 and StreptomycesStreptomyces collinusCGMCC 4.1623 T The similarity of (2) was 98.7%. Phylogenetic analysis also showed the strain SCSIO K001 and StreptomycesS. collinusCGMCC 4.1623 T The evolutionary distance is closest (as shown in fig. 2). In addition, the strain SCSIO K001 and the most similar speciesS. collinusCGMCC 4.1623 T The average nucleotide homology (ANI) and DNA-DNA hybridization (dDDH) values were 82.5% and 26.2%, respectively, well below the standard threshold for division of prokaryotic microorganism species (internationally recognized inter-species discrimination threshold ANI 95% -96%; dDDH 70%). It can thus be determined as a new species of Streptomyces.
The 16S rRNA gene sequence of the strain SCSIO K001 is shown as SEQ ID NO.1, and specifically comprises the following steps:
>SCSIO K001
ACCCGTCCCCTTCGAACAGCTCCCTCCCCGAGGGGTTGGGCCACCGGCTTCGGGTGTTACCGACTTTCGTGACGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCAGCAATGCTGATCTGCGATTACTAGCGACTCCGACTTCATGGGGTCGAGTTGCAGACCCCAATCCGAACTGAGACCGGCTTTTTGAGATTCGCTCCACCTCACGGTATCGCAGCTCATTGTACCGGCCATTGTAGCACGTGTGCAGCCCAAGACATAAGGGGCATGATGACTTGACGTCGTCCCCACCTTCCTCCGAGTTGACCCCGGCGGTCTCCCGTGAGTCCCCAGCACCACAAGGGCCTGCTGGCAACACGGGACAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAGCCATGCACCACCTGTACACCGACCACAAGGGGGGCACTATCTCTAATGCTTTCCGGTGTATGTCAAGCCTTGGTAAGGTTCTTCGCGTTGCGTCGAATTAAGCCACATGCTCCGCCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTTAGCCTTGCGGCCGTACTCCCCAGGCGGGGCACTTAATGCGTTAGCTGCGGCACGGACAACGTGGAATGTTGCCCACACCTAGTGCCCACCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACGCTTTCGCTCCTCAGCGTCAGTATCGGCCCAGAGATCCGCCTTCGCCACCGGTGTTCCTCCTGATATCTGCGCATTTCACCGCTACACCAGGAATTCCGATCTCCCCTACCGAACTCTAGCCTGCCCGTATCGACTGCAGACCCGGGGTTAAGCCCCGGGCTTTCACAACCGACGCGACAAGCCGCCTACGAGCTCTTTACGCCCAATAATTCCGGACAACGCTTGCGCCCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGGCGCTTCTTCTGCAGGTACCGTCACTCTCGCTTCTTCCCTGCTGAAAGAGGTTTACAACCCGAAGGCCGTCATCCCTCACGCGGCGTCGCTGCATCAGGCTTGCGCCCATTGTGCAATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGGTCGCCCTCTCAGGCCGGCTACCCGTCGTCGCCTTGGTGAGCCATTACCTCACCAACAAGCTGATAGGCCGCGGGCTCATCCTGCACCGCCGGAGCTTTCGAACCCTCTGGATGCCCAGAAGGATCAGTATCCGGTATTAGACCCCGTTTCCAGGGCTTGTCCCAGAGTGCAGGGCAGATTGCCCACGTGTTACTCACCCGTTCGCCACTAATCCCCACCGAAGTGGTTCATCGTTCGACTTGCATGGT。
thus, the strain SCSIO K001 of the present invention is assigned toStreptomycesNamed streptomycete [ (]Streptomycessp.) SCSIO K001 deposited at China general microbiological culture Collection center (CGMCC) at a position of 2023, 07, 05: the institute of microbiology, national institute of sciences, no. 3, north chen west way 1, region of korea, beijing city, postal code: 100101 and the preservation number is CGMCC No.27800.
Example 2: application of streptomycete SCSIO K001 in production of anthrax mycin and beta-erythromycin
(1) Preparation of fermentation strains
The strain SCSIO K001 is inoculated into a seed culture medium and cultured for 48 hours at 28 ℃ to obtain seed liquid. The formula of the seed culture medium is as follows: 10 g/liter of soluble starch, K 2 HPO 4 1 g，MgSO 4 ·7H 2 O 1 g，(NH 4 ) 2 SO 4 2 g，CaCO 3 2 g,10 times trace element solution 1 mL, peptone 1 g, yeast extract 0.5 g, sea salt 30 g, water as solvent and pH 7.2-7.4 (seed culture medium is prepared by dissolving each component in water, stirring and dissolving, regulating pH, sterilizing and cooling); the formula of the 10 x trace element solution is as follows: 1.5 g/liter of ferrous oxide, 10 mL g/liter of 7.7M hydrochloric acid, 0.006 g g/liter of cobalt chloride, 0.036 g g/liter of manganese chloride, 0.024. 0.024 g g/liter of zinc chloride, 0.002. 0.002 g g/liter of boric acid, 0.19. 0.19 g g/liter of sodium molybdate, 0.1. 0.1 g g/liter of nickel chloride, 0.07 g/g g/liter of copper chloride, water as solvent and 6.0 pH.
(2) Fermenting to produce anthracycline and beta-Yuzhithromycin by using the strain obtained above
Inoculating the seed solution into a fermentation culture medium according to the volume ratio of 1:20, and culturing at 28 ℃ for 200 hours to obtain a streptomyces SCSIO K001 fermentation culture containing anthrax mycin and beta-erythromycin. The formula of the fermentation medium is as follows: yeast extract 4 g/L, malt extract 10 g/L, glucose 4 g/L, sea salt 30 g/L, and water as solvent (fermentation medium is prepared by dissolving each component in water, stirring for dissolving, sterilizing, and cooling).
(3) Extraction and separation of anthrax and beta-Yuzhixin from fermentation culture
(a) Separating the fermentation broth and mycelium of the fermentation culture, extracting the fermentation broth by butanone, and concentrating the butanone extract to obtain extract A; extracting mycelium with acetone, concentrating the acetone extract to obtain extract B;
(b) Subjecting the crude extract A and extract B to gradient elution with chloroform-methanol (10:0-5:5, volume ratio) by 100-200 mesh silica gel column, and detecting each fraction by thin layer chromatography and HPLC-UV. Fraction B (chloroform: methanol=9:1 volume fraction eluted) was subjected to medium pressure column chromatography and eluted with a methanol/water (100:0-10:90 volume ratio) gradient to give 10 fractions B1-B10. Fraction B2 (methanol: water=90:10 volume fraction) was separated by Sephadex LH-20 column (mobile phase 100% methanol) and semi-preparative High Performance Liquid Chromatography (HPLC) to give anthramycin (1, t) R =36.1 min, 4.2 mg). Fraction B1 (100% methanol elution fraction) was purified by Sephadex LH-20 column (mobile phase dichloromethane: methanol=1:1 volume ratio) and by semi-preparative HPLC elution to give β -erythromycin (2, t) R =19.8 min, 3.7 mg）。
(4) Spectral data
Anthrax mycin: c (C) 25 H 32 O 4 , ESI-MSm/z397.7 [M+H] + , 1 H NMR (500 MHz, Chloroform-d)δ6.51 (m, 1H), 5.98 (s, 1H), 5.92 (t,J= 10.8 Hz, 1H), 5.75 (d,J= 10.1 Hz, 1H), 5.59 (overlapped, 3H), 5.45 (t,J= 9.5 Hz, 1H), 5.41 (d,J= 3.5 Hz, 1H), 3.58 (q,J = 6.9 Hz, 1H), 2.63 (overlapped, 3H), 2.42 (m, 1H), 2.06 (m, 1H), 2.01 (m, 1H), 1.95 (m, 1H), 1.84 (m, 1H), 1.71 (s, 3H), 1.55 (overlapped, 1H), 1.44 (d,J= 6.9 Hz, 3H), 1.37 (d,J= 6.7 Hz, 3H), 0.98 (d,J= 6.5 Hz, 3H) 。
Beta-erythromycin: c (C) 27 H 20 O 12 , ESI-MSm/z537.2 [M+H] + , 1 H NMR (500 MHz, Chloroform-d)δ 7.47 (s, 1H), 6.95 (s, 1H), 6.78 (s, 1H), 4.04 (s, 3H), 4.03 (s, 3H), 3.99 (s, 3H), 3.68 (d,J= 18.9 Hz, 1H), 3.45 (m, 1H), 3.35 (d,J= 18.9 Hz, 1H), 3.01 (m, 1H), 2.51 (dd,J= 13.9, 5.8 Hz, 1H), 2.31 (m, 1H) 。
The chemical structures of anthrax mycin (1) and beta-jade erythromycin (2) are shown in figure 3.
The foregoing is merely a preferred embodiment of the present invention, and it should be noted that the above-mentioned preferred embodiment should not be construed as limiting the invention, and the scope of the invention should be defined by the appended claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and such modifications and adaptations are intended to be comprehended within the scope of the invention.

Claims (8)

1. Novel marine streptomycete for producing anthrax mycin and beta-erythromycinStreptomycessp.) SCSIO K001 with deposit number: CGMCC No.27800.
2. The Streptomyces species of claim 1Streptomycessp.) SCSIO K001 in the production of anthrax and beta-jade erythromycin.
3. A process for preparing Streptomyces sp according to claim 1Streptomycessp.) method for producing anthracycline and beta-erythromycin by SCSIO K001, comprising the steps of: preparing a fermentation culture of streptomyces SCSIO K001; separating fermentation broth and mycelium of the fermentation culture, extracting the fermentation broth by butanone, and concentrating the butanone extract to obtain extract A; extracting mycelium with acetone, concentrating the acetone extract to obtain extract B; and (3) purifying the crude extract obtained by combining the extract A and the extract B through silica gel column chromatography and semi-preparative HPLC to obtain the compounds anthracnose mycin and beta-Yuerythromycin.
4. A method according to claim 3, wherein the fermentation culture is prepared by: and inoculating streptomyces SCSIO K001 into a seed culture medium for culturing for 24-48 hours to obtain seed liquid, and inoculating the seed liquid into a fermentation culture medium for culturing for 144-240 hours to obtain a fermentation culture.
5. The method according to claim 4, wherein the optimal culture temperature of Streptomyces SCSIO K001 is 28 ℃; the seed liquid is inoculated according to the volume ratio of 1:20.
6. The method of claim 4, wherein the seed medium is formulated as follows: 10 g/liter of soluble starch, K 2 HPO 4 1 g，MgSO 4 ·7H 2 O 1 g，(NH 4 ) 2 SO 4 2 g，CaCO 3 2 g of 10 times trace element solution 1 mL, peptone 1 g, yeast extract 0.5 g and sea salt 30 g, wherein the solvent is water, and the pH is 7.2-7.4;
the formula of the 10 x trace element solution is as follows: 1.5 g/liter of ferrous oxide, 10 mL M hydrochloric acid, 0.006 g M cobalt chloride, 0.036 g M manganese chloride, 0.024. 0.024 g M zinc chloride, 0.002 g boric acid, 0.19 g M sodium molybdate, 0.1. 0.1 g M nickel chloride and 0.07 g M copper chloride, water as solvent and pH 6.0.
7. The method of claim 4, wherein the fermentation medium is formulated as follows: yeast extract 4 g/L, malt extract 10 g/L, glucose 4 g/L and sea salt 30 g/L, and water as solvent.
8. The Streptomyces species of claim 1Streptomycessp.) application of SCSIO K001 in preparing antibacterial agent for inhibiting Staphylococcus aureus, bacillus subtilis and Aspergillus niger; the bacteriostatic agent takes the streptomycete SCSIO K001 or a fermentation culture thereof as an active ingredient.
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101974464A (en) * 2010-10-18 2011-02-16 中国科学院南海海洋研究所 Streptomyces and process for preparing antimycin antibiotics by fermentation using same
CN112409372A (en) * 2020-11-05 2021-02-26 哈药慈航制药股份有限公司 Rubicin analogue, preparation method and application thereof

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11028113B2 (en) * 2018-10-26 2021-06-08 Wisconsin Alumni Research Foundation Cyphomycin, compositions and uses thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101974464A (en) * 2010-10-18 2011-02-16 中国科学院南海海洋研究所 Streptomyces and process for preparing antimycin antibiotics by fermentation using same
CN112409372A (en) * 2020-11-05 2021-02-26 哈药慈航制药股份有限公司 Rubicin analogue, preparation method and application thereof

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
Anthracimycin B, a Potent Antibiotic against Gram-Positive Bacteria Isolated from Cultures of the Deep-Sea Actinomycete Streptomyces cyaneofuscatus M-169;Víctor Rodríguez et al.;marine drugs;第16卷(第11期);第1-8页 *
Biosynthesis of the Novel Macrolide Antibiotic Anthracimycin;Silke Alt and Barrie Wilkinson;ACS Chem. Biol.;第10卷(第11期);第2468−2479页 *
Biosynthetic origin of anthracimycin: a tricyclic macrolide from Streptomyces sp.;Enjuro Harunari et al.;The Journal of Antibiotics;第69卷;第403–405页 *
Hyaluromycin, a New Hyaluronidase Inhibitor of Polyketide Origin from Marine Streptomyces sp.;Enjuro Harunari et al.;marine drugs;第12卷(第1期);第491-507页 *
Partial Activation of a Silent Angucycline-type Gene Cluster from a Rubromycin β Producing Streptomyces sp. PGA64;MIKKO METSA-KETELA et al.;THE JOURNAL OF ANTIBIOTICS;第57卷(第8期);第502-510页 *

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