CN114703111B - Streptomyces hydrogenogenes ZJPH2021031 and application thereof in preparation of antibacterial agent - Google Patents
Streptomyces hydrogenogenes ZJPH2021031 and application thereof in preparation of antibacterial agent Download PDFInfo
- Publication number
- CN114703111B CN114703111B CN202210515031.5A CN202210515031A CN114703111B CN 114703111 B CN114703111 B CN 114703111B CN 202210515031 A CN202210515031 A CN 202210515031A CN 114703111 B CN114703111 B CN 114703111B
- Authority
- CN
- China
- Prior art keywords
- streptomyces
- fermentation
- strain
- hydrogenans
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmacology & Pharmacy (AREA)
- Zoology (AREA)
- Animal Behavior & Ethology (AREA)
- Wood Science & Technology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Microbiology (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- General Chemical & Material Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to streptomyces hydrogenised ZJPH2021031 and application thereof in preparation of antibacterial agents, the strain has stronger antibacterial activity, especially has strongest antibacterial activity on staphylococcus aureus, and provides a new microorganism source and a new path for preparation of the antibacterial agents. The antibacterial activity of the fermentation liquor of the obtained strain is further obviously improved through optimizing the fermentation medium.
Description
Technical Field
The invention relates to a Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031 and application thereof in preparation of antibacterial agents, and belongs to the technical field of bioengineering.
Background
Due to the irregular use of antibiotics and the ever-increasing resistance of pathogenic microorganisms, there is an increasing need to find new compounds of microbial origin with antibacterial activity. Actinomycetes have been found to have significant advantages in the production of various biologically active secondary metabolites, such as the antibacterial agent daptomycin and the anticancer molecule, r Bei Kamei, etc., both from actinomycetes. However, since known compounds are often obtained when the isolation of natural products from the same or similar soil microorganisms is repeated for a long period of time, the possibility of obtaining antibacterial natural products having a novel skeleton structure is greatly reduced. In recent years, with respect to microorganisms in marine environments, research has been focused on: (1) Deep sea sediments are present in coastal and offshore areas (2), and it has been noted that the density of microbial populations such as actinomycetes is higher in shallow sea areas than in deep sea areas. This patent is through separating actinomycetes from the coastal saline soil that gathers from near the mouth of the hantaan qiangtang river, and look for novel natural bacteriostat from it.
Disclosure of Invention
The invention aims to provide Streptomyces hydrogeneratus ZJPH2021031 which is screened from Hangzhou bay seashore saline soil type soil and can produce antibacterial active substances, and application of the antibacterial active substances produced by the Streptomyces hydrogeneratus ZJPH2021031 in bacteriostasis.
In order to achieve the above purpose, the invention adopts the following technical scheme:
in a first aspect, the present invention provides a strain of Streptomyces hydrogenogenes (Streptomyces hydrogenans)
ZJPH2021031, deposited in chinese collection of typical cultures, accession number: cctccc NO: m2022490, date of preservation: 2022, 4, 25 days, address: 430072, university of martial arts, wuhan, china.
In a second aspect, the invention provides the use of the strain Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031 described above for the preparation of an antibacterial agent having antibacterial activity.
Further, the bacteria used for the evaluation of antibacterial activity in the present invention are Staphylococcus aureus (Staphylococcus aureus), bacillus subtilis (Bacillus subtilis), escherichia coli (Escherichia coli) or Pseudomonas aeruginosa (Pseudomonas aeruginosa).
In a specific embodiment of the invention, the application is: the fermentation broth obtained by fermenting and culturing the streptomyces hydrogenative (Streptomyces hydrogenans) ZJPH2021031 (culturing at 30 ℃) is applied to preparing an antibacterial agent with antibacterial activity.
The fermentation culture process specifically comprises the following steps: inoculating the streptomyces hydrogenated (Streptomyces hydrogenans) ZJPH2021031 into an ISPII seed culture medium, performing shaking culture at 30 ℃ for 24 hours, inoculating the streptomyces hydrogenated (Streptomyces hydrogenans) into a fermentation culture medium with an inoculum size of 5% (v/v), performing shaking culture at 30 ℃ for 24 hours, and centrifuging to obtain the fermentation broth.
Preferably, the final concentration composition of the fermentation medium used for the fermentation culture is as follows: 10.0 to 15.0g/L of glycerin, 0.2 to 0.8g/L of L-tyrosine, 0.8 to 1.3g/L of L-asparagine and K 2 HPO 4 0.2～0.8g/L，MgSO 4 ·7H 2 O0.3～0.9g/L，NaCl 0.2～1.0g/L，FeSO 4 ·7H 2 O0.01-0.02 g/L, water as solvent and pH 7.0-7.5.
Preferably, the final concentration composition of the fermentation medium is as follows: 15g/L of glycerol, 0.5g/L of L-tyrosine, 1.0g/L of L-asparagine and K 2 HPO 4 0.5g/L,MgSO 4 ·7H 2 O 0.5g/L,NaCl 0.5g/L,FeSO 4 ·7H 2 O0.01 g/L, water as solvent and pH 7.2-7.4.
Further, the final concentration composition of the ISPII seed medium is as follows: yeast extract 4.0g/L, wort 10.0g/L, glucose 4.0g/L, water as solvent, and pH 7.2-7.4.
The Streptomyces hydrogeneratus (Streptomyces hydrogenans) ZJPH2021031 strain is separated from Hangzhou bay seashore saline soil, and fermentation liquor of the strain has strong antibacterial activity.
The bacterial colony of the separated streptomyces hydrogenated (Streptomyces hydrogenans) ZJPH2021031 strain is opaque and bulges on a culture medium, has clear outline, is not easy to pick up, is unordered fiber-shaped when the hypha is observed under a microscope, belongs to gram positive bacteria, can grow in the temperature range of 5-40 ℃, and has the optimal growth temperature of 26-30 ℃. The 16S rRNA sequence of the Streptomyces hydrogenosis (Streptomyces hydrogenans) ZJPH2021031 strain is shown in seq. NO1.
The invention also provides the antibacterial activity of the fermentation liquor of the Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031 strain. Streptomyces hydrogenization (Streptomyces hydrogenans)
The fermentation liquor of ZJPH2021031 strain has stronger antibacterial activity, especially shows antibacterial activity higher than 50mM streptomycin sulfate for staphylococcus aureus, and has strong antibacterial capability.
The invention screens the optimal fermentation medium composition of the Streptomyces hydrogenans (Streptomyces hydrogenans) ZJPH2021031 strain, and discovers that the fermentation broth has higher antibacterial activity after fermentation culture by the medium composed of the following components.
Medium M6: 15.0g/L of glycerol, 0.5g/L of L-tyrosine, 1.0g/L of L-asparagine, K 2 HPO 4 0.5g/L，MgSO 4 ·7H 2 O 0.5g/L，NaCl 0.5g/L，FeSO 4 ·7H 2 O 0.01g/L, water as solvent and pH 7.2-7.4.
The Streptomyces hydrogenization (Streptomyces hydrogenans) ZJPH2021031 strain has stronger antibacterial activity on fermentation broth obtained by fermentation culture, and particularly has stronger antibacterial activity on staphylococcus aureus. Thus, the strain can be used for preparing the corresponding antibacterial agent by fermentation.
Compared with the prior art, the strain capable of producing the antibacterial active substance through fermentation is a novel strain separated from the Hangzhou bay seashore solonchak soil by the inventor, the strain is named as Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031 after identification, the fermentation broth obtained through fermentation culture has stronger antibacterial activity, particularly has better antibacterial activity on staphylococcus aureus, a novel microorganism strain and a novel path can be provided for preparing a novel antibacterial agent, and the antibacterial activity of the fermentation broth of the strain can be remarkably improved through optimizing a fermentation medium of the strain.
Drawings
FIG. 1 is a phylogenetic tree (N-J method) of Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031 strain
FIG. 2 shows colony morphology of Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031 strain
FIG. 3 is a photomicrograph (400X) of a strain of Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031
Detailed Description
The technical scheme of the invention is further specifically described through specific embodiments.
Example 1
The coastal saline soil used in this example was collected from soil in the dike of the Qian pond, out of the sea, near the Hangzhou bay (121.189 ° east, 30.365 ° north latitude) in the Yangtze river, cixi county, ningbo, zhejiang.
The screening culture medium adopted in the embodiment is a composite culture medium containing potassium dichromate and KNO, and the weight ratio of each component is respectively that soluble starch is 20.0g/L and KNO 3 1.0g/L，NaCl 0.5g/L，Fe(SO 4 ) 3 ·7H 2 O 0.01g/L，MgSO 4 0.24g/L，3H 2 O·K 2 HPO 4 0.752g/L, 20.0g/L of agar powder, 0.075g/L of potassium dichromate, water as solvent, and pH value of 7.2-7.4.
Screening method of Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031 strain of this example:
drying collected Hangzhou Baside saline soil at 120deg.C for 2 hr, weighing 5g of dry soil, adding into 0.9% (w/v) sterile physiological saline, oscillating at 30deg.C for 30min, and gradient diluting with sterile water to 10 -2 ～10 -5 . 100 mu L of bacterial suspension is absorbed and coated on a first Gao's synthetic culture medium containing 0.0075% (w/v) potassium dichromate, screening culture is carried out for 7 days at 30 ℃, then the culture is transferred to a flat plate for culture, single bacterial colonies with different forms are picked up and transferred to a first Gao's culture medium flat plate for 3-5 times for culture until a single bacterial colony pure culture is formed, and finally an actinomycete strain is obtained. The bacterial colony of the strain is micro-bulge, is tightly combined with a culture medium and is not easy to pick, white spores are gathered on the surface and are in a crater shape, and the bacterial colony is marked as a strain ZJPH2021031.
Example 2
Molecular biological identification of the strain ZJPH2021031 of the present invention, namely PCR amplification of 16S rDNA was performed on the strain ZJPH2021031 of the present invention by a method conventional in the art. More specifically, the whole genome of the strain was extracted using a bacterial DNA extraction kit of the division of biological engineering (Shanghai), and then PCR was performed using a universal primer 27F (AGTTTGATCMTGGCTCAG), 1492R (: GGTTACCTTGTTACGACTT) synthesized by the division of biological engineering (Shanghai), the PCR procedure being pre-denatured for 5min 1cycle at 94 ℃; denaturation at 94℃for 30s, annealing at 54℃for 30s, extension at 72℃for 1min,35cycles; extending at 72 ℃ for 10min 1cycle; preserving heat at 4 ℃. The PCR product was purified by a PCR product purification kit provided by the Protoengineering (Shanghai) Co., ltd and submitted to sequencing by the Protoengineering (Shanghai) Co., ltd. The sequencing results are shown as seq. NO1. The 16S rDNA sequence of the strain ZJPH2021031 of the present invention was subjected to homology alignment with the EzBioCloud' S typical strain database using Blast alignment, and the result showed that the 16S rDNA sequence of the strain ZJPH2021031 had a sequence similarity of 98.45% with Streptomyces hydrogenogenes (Streptomyces hydrogenans) JCM 4771, and then a phylogenetic tree of the strain ZJPH2021031 was constructed by MEGA (V7.0) using the Neighbor-Joining, NJ), and the result showed that the strain ZJPH2021031 of the present invention had 87% homology with the strain Streptomyces hydrogenans JCM 4771, and the strain ZJPH2021031 was determined to be Streptomyces sp according to the comparison of gene homology, named Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJ 2021031, deposited in the China center for type culture collection, accession number: cctccc M2022490, with a date of preservation of 2022, 4 months and 25 days.
Example 3
The strain Streptomyces hydrogenans ZJPH2021031 obtained by screening in the method of example 1 was selected, inoculated into a first high-grade synthetic solid medium, obliquely inserted into a sterilized cover glass at 45 degrees, and cultured upside down at 30 ℃ for 7 days, and the colony morphology and mycelium growth were observed. The morphological characteristics and the physiological and biochemical characteristic results of the strain ZJPH2021031 show that: as shown in FIG. 2, the bacterial colony of the strain ZJPH2021031 of the present invention is opaque and has protrusions and folds, has a rough surface, has white spores, has a clear colony outline, is not easy to pick up, and has a mycelium growth form as shown in FIG. 3, and the mycelium growth form is observed under a microscope (400×), so that aerial hyphae are thick and sparse, and the hyphae in the basal group are thin and dense. In addition, the streptomyces hydrogenated (Streptomyces hydrogenans) ZJPH2021031 strain belongs to gram-positive bacteria, can grow in the temperature range of 5-40 ℃, and has the optimal growth temperature of 26-30 ℃.
Example 4
Antibacterial Activity of Streptomyces hydrogeneratus (Streptomyces hydrogenans) ZJPH2021031 Strain fermentation broth according to the present invention
The bacteriostatic activity indicating strain described in this example was selected from Staphylococcus aureus (Staphylococcus aureus), bacillus subtilis (Bacillus subtilis), escherichia coli (Escherichia coli), pseudomonas aeruginosa (Pseudomonas aeruginosa).
The seed culture medium ISPII used in this example was prepared as follows: yeast extract 4.0g/L, wort 10.0g/L, glucose 4.0g/L, water as solvent, and pH 7.2-7.4.
The weight ratio of the fermentation medium M1 used in this example was as follows: yeast extract 4.0g/L, wort 10.0g/L, glucose 4.0g/L, water as solvent, and pH 7.2-7.4.
The separated strain ZJPH2021031 of streptomyces hydrogenans (Streptomyces hydrogenans) is inoculated into an ISPII seed culture medium, after shaking culture is carried out for 24 hours at 30 ℃, the strain is inoculated into 100mL of an M1 fermentation culture medium with an inoculum size of 5% (v/v), shaking culture is carried out for 24 hours at 30 ℃, fermentation broth is obtained by centrifugation, after thalli are removed by centrifugation, 10 mu L of filtered and sterilized fermentation broth sample is dripped on a filter paper sheet, 10 mu L of filtered and sterilized 50mM streptomycin sulfate aqueous solution and sterile water are respectively used as a positive control and a negative control, and after standing culture is carried out for 24 hours in a constant temperature incubator at 37 ℃, the antibacterial activity of the strain Streptomyces hydrogenans ZJPH2021031 fermentation broth is obtained by observing and measuring the antibacterial activity of the strain.
The concrete description is as follows:
A. inoculating Streptomyces hydrogenans (Streptomyces hydrogenans) ZJPH2021031 strain subjected to plate activation culture into an ISPII seed culture medium, performing shake culture at 30 ℃ for 24 hours, inoculating into 100mL of an M1 fermentation culture medium with an inoculum size of 5% (v/v), performing shake culture at 30 ℃ for 24 hours, centrifuging to obtain fermentation liquor, centrifuging to remove thalli, and preserving at 4 ℃ for later use;
B. inoculating staphylococcus aureus, escherichia coli, bacillus subtilis and pseudomonas aeruginosa subjected to plate activation culture into 50mL of LB liquid culture medium, and shake culturing at 37 ℃ and 180rpm for 12 hours;
C. taking 2mL of the culture solution prepared in the step B, and measuring the OD of the corresponding culture solution by using a spectrophotometer 600 Numerical value, and diluting the concentration of the bacterial liquid to OD by using sterile water 600 Values in the range of 0.7-0.8 (ensuring consistent numbers of experimental strains accessed in subsequent repeated experiments);
D. mu.L of the bacterial suspension was applied to a solid LB plate and uniformly coated with a coating bar. Culturing in a constant temperature incubator at 37deg.C for 30min. Taking out the plates, uniformly putting 5 sterilized circular filter paper sheets into each plate, and lightly pressing the filter paper sheets to ensure that the filter paper sheets are in full contact with the solid culture medium.
E. According to the set sample adding sequence, 10 mu L of the fermentation broth prepared in the step A is filtered and sterilized, and then is dripped into the middle part of a round filter paper sheet (the diameter of the filter paper sheet is 6 mM), and simultaneously, two filter paper sheets added with sterile water with the same volume as the fermentation broth and 50mM streptomycin sulfate aqueous solution after filtration sterilization are respectively arranged in each solid flat plate containing LB culture medium to serve as a negative control and a positive control.
F. The solid plates were then incubated at 37℃for 24 hours, and the size of the zone of inhibition was observed and measured to evaluate the antibacterial activity of the fermentation broth of Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031.
Example 5
The cultivation and fermentation process of the Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031 strain producing an antibacterial active material in this example is the same as that of example 4, except that:
the fermentation medium used in this example is M2 medium, and the ratio of the components of the medium is: 10.0g/L tryptone, 5.0g/L yeast extract, 10.0g/L NaCl, water as solvent and pH 7.0-7.4. Other operation steps are the same as in embodiment 4, and will not be repeated here.
Example 6
The cultivation and fermentation process of the Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031 strain producing an antibacterial active material in this example is the same as that of example 4, except that:
the fermentation medium used in this example is M3 medium, and the ratio of the components of the medium is: glucose 40.0g/L, peptone 10.0g/L, water as solvent, and pH 5.8-6.2.
Other operation steps are the same as in embodiment 4, and will not be repeated here.
Example 7
The cultivation and fermentation process of the Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031 strain producing an antibacterial active material in this example is the same as that of example 4, except that:
the fermentation medium used in this example is M4 medium, and the ratio of the components of the medium is:
M4:K 2 HPO 4 2.0g/L，MgSO 4 ·7H 2 O 2.0g/L，NaCl 2.0g/L，(NH 4 ) 2 SO 4 4.0g/L，CaCO 3 4.0g/L, water as solvent and pH 7.0-7.4.
Other operation steps are the same as in embodiment 4, and will not be repeated here.
Example 8
The cultivation and fermentation process of the Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031 strain producing an antibacterial active material in this example is the same as that of example 4, except that:
the fermentation medium used in this example is M5 medium, and the ratio of the components of the medium is: l-asparagine 1.0g/L, glycerol 10.0g/L, K 2 HPO 4 1.0g/L, water as solvent and pH 7.2-7.5. Other operation steps are the same as in embodiment 4, and will not be repeated here.
Example 9
The culture and fermentation methods for producing Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031 strain with antibacterial activity in this example were the same as those of example 4, except that:
the fermentation medium used in this example is M6 medium, and the ratio of the components of the medium is: 15g/L of glycerol, 0.5g/L of L-tyrosine, 1.0g/L of L-asparagine and K 2 HPO 4 0.5g/L，MgSO 4 ·7H 2 O 0.5g/L，NaCl 0.5g/L，FeSO 4 ·7H 2 O0.01 g/L, water as solvent and pH 7.2-7.4.
Other operation steps are the same as in example 4 and will not be repeated here
TABLE 1 bacteriostatic Activity of fermentation broths obtained after Streptomyces hydrogenus (Streptomyces hydrogenans) ZJPH2021031 were cultured in different fermentation media
Streptomycin sulfate was used as a positive control; sterile water was used as a negative control; the unit of the inhibition zone is millimeter (mm)
As shown in Table 1, the fermentation broth obtained after the strain Streptomyces hydrogenans ZJPH2021031 is cultured has obvious antibacterial effect on bacillus subtilis, staphylococcus aureus, pseudomonas aeruginosa and escherichia coli, wherein the fermentation broth shows optimal antibacterial activity on staphylococcus aureus, which indicates that natural compounds capable of obviously inhibiting staphylococcus aureus are generated in the fermentation broth of Streptomyces hydrogenans ZJPH2021031 strain, and the strain has application value in developing novel antibacterial agents for drug-resistant staphylococcus aureus. From table 1, it is also known that the strain ZJPH2021031 exhibits higher bacteriostatic activity against gram-positive bacteria in the M6 fermentation medium relative to the initial fermentation medium M1 (ISP II), and thus the optimal fermentation medium type for producing the antibacterial agent is M6. From the results, streptomyces hydrogenans ZJPH2021031 strain is cultured in a culture medium containing a plurality of metal ions by using amino acid as a nitrogen source, and the obtained fermentation broth has stronger antibacterial activity.
Sequence listing
<110> Zhejiang university of industry
<120> Streptomyces hydrogeneratus ZJPH2021031 and use thereof in preparation of antibacterial agent
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1422
<212> DNA
<213> Streptomyces hydrogeneratus ZJPH2021031 (Streptomyces hydrogenans ZJPH 2021031)
<400> 1
ggatgggggg gtctacatgc agtcgacgat gagcccattc ggggtggatt agtggcgaac 60
gggtgagtaa cacgtgggca atctgccctt cactctggga caagccctgg aaacggggtc 120
taataccgga tacgagttcg ggaggcatct cctggactgg aaagctccgg cggtgaagga 180
tgagcccgcg gcctatcagc ttgttggtgg ggtaatggcc taccaaggcg acgacgggta 240
gccggcctga gagggcgacc ggccacactg ggactgagac acggcccaga ctcctacggg 300
aggcagcagt ggggaatatt gcacaatggg cgaaagcctg atgcagcgac gccgcgtgag 360
ggatgacggc cttcgggttg taaacctctt tcagcaggga agaagcgaaa gtgacggtac 420
ctgcagaaga agcgccggct aactacgtgc cagcagccgc ggtaatacgt agggcgcaag 480
cgttgtccgg aattattggg cgtaaagagc tcgtaggcgg cttgtcacgt cgggtgtgaa 540
agcccggggc ttaaccccgg gtctgcatcc gatacgggca ggctagagtg tggtagggga 600
gatcggaatt cctggtgtag cggtgaaatg cgcagatatc aggaggaaca ccggtggcga 660
aggcggatct ctgggccatt actgacgctg aggagcgaaa gcgtggggag cgaacaggat 720
tagataccct ggtagtccac gccgtaaacg ttgggaacta ggtgttggcg acattccacg 780
tcgtcggtgc cgcagctaac gcattaagtt ccccgcctgg ggagtacggc cgcaaggcta 840
aaactcaaag gaattgacgg gggcccgcac aagcagcgga gcatgtggct taattcgacg 900
caacgcgaag aaccttacca aggcttgaca tataccggaa agcattagag atagtgcccc 960
ccttgtggtc ggtatacagg tggtgcatgg ctgtcgtcag ctcgtgtcgt gagatgttgg 1020
gttaagtccc gcaacgagcg caacccttgt cctgtgttgc cagcatgccc ttcggggtga 1080
tggggactca caggagaccg ccggggtcaa ctcggaggaa ggtggggacg acgtcaagtc 1140
atcatgcccc ttatgtcttg ggctgcacac gtgctacaat ggccggtaca aagagctgcg 1200
atgccgcgag gcggagcgaa tctcaaaaag ccggtctcag ttcggattgg ggtctgcaac 1260
tcgaccccat gaagtcggag ttgctagtaa tcgcagatca gcattgctgc ggtgaatacg 1320
ttcccgggcc ttgtacacac cgcccgtcac gtcacgaaag tcggtaacac ccgaagccgg 1380
tggtcccaac cccttgtgga ggagctcgaa cattccgggc cc 1422
Claims (9)
1. Streptomyces hydrogenogenes (Streptomyces hydrogenans) ZJPH2021031, deposited with China center for type culture Collection, accession number: cctccc NO: m2022490, date of preservation: 2022, 4, 25 days, address: 430072, university of martial arts, wuhan, china.
2. Use of streptomyces hydrogenated (Streptomyces hydrogenans) ZJPH2021031 according to claim 1 for the preparation of an antibacterial agent having antibacterial activity, characterized in that: the bacteria are staphylococcus aureus, bacillus subtilis, escherichia coli or pseudomonas aeruginosa.
3. The use according to claim 2, wherein: the bacteria are staphylococcus aureus.
4. The application according to claim 2, characterized in that it is: and (3) applying fermentation broth obtained by fermenting and culturing the streptomyces hydrogenative (Streptomyces hydrogenans) ZJPH2021031 to prepare an antibacterial agent.
5. The use according to claim 4, wherein: the temperature of the fermentation culture is 30 ℃.
6. The use according to claim 4, wherein the fermentation culture is operated as follows: inoculating the streptomyces hydrogenated (Streptomyces hydrogenans) ZJPH2021031 into an ISPII seed culture medium, performing shaking culture at 30 ℃ for 24 hours, inoculating the streptomyces hydrogenated (Streptomyces hydrogenans) into a fermentation culture medium with the volume inoculum size of 5%, performing shaking culture at 30 ℃ for 24 hours, and centrifuging to obtain the fermentation liquor.
7. The use according to claim 6, characterized in that the final concentration composition of the fermentation medium is as follows: 10.0 to 15.0g/L of glycerin, 0.2 to 0.8g/L of L-tyrosine and 0.8 to 1g/L of L-asparagine.3g/L，K 2 HPO 4 0.2～0.8g/L，MgSO 4 ·7H 2 O 0.3～0.9g/L，NaCl 0.2～1.0g/L，FeSO 4 ·7H 2 O0.01-0.02 g/L, water as solvent and pH 7.0-7.5.
8. The use according to claim 7, characterized in that the final concentration composition of the fermentation medium is as follows: 15g/L of glycerol, 0.5g/L of L-tyrosine, 1.0g/L of L-asparagine and K 2 HPO 4 0.5g/L,MgSO 4 ·7H 2 O0.5g/L,NaCl 0.5g/L,FeSO 4 ·7H 2 O0.01 g/L, water as solvent and pH 7.2-7.4.
9. The use according to claim 6, wherein: the final concentration composition of the ISPII seed culture medium is as follows: yeast extract 4.0g/L, wort 10.0g/L, glucose 4.0g/L, water as solvent, and pH 7.2-7.4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210515031.5A CN114703111B (en) | 2022-05-11 | 2022-05-11 | Streptomyces hydrogenogenes ZJPH2021031 and application thereof in preparation of antibacterial agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210515031.5A CN114703111B (en) | 2022-05-11 | 2022-05-11 | Streptomyces hydrogenogenes ZJPH2021031 and application thereof in preparation of antibacterial agent |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114703111A CN114703111A (en) | 2022-07-05 |
| CN114703111B true CN114703111B (en) | 2023-08-08 |
Family
ID=82176770
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210515031.5A Active CN114703111B (en) | 2022-05-11 | 2022-05-11 | Streptomyces hydrogenogenes ZJPH2021031 and application thereof in preparation of antibacterial agent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114703111B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103370412A (en) * | 2010-12-04 | 2013-10-23 | 万芬 | RNA stability enhancer |
| CN110218671A (en) * | 2019-05-29 | 2019-09-10 | 江苏师范大学 | One plant of production water streptomycete and its application in biological control |
| CN114231435A (en) * | 2021-10-14 | 2022-03-25 | 商丘师范学院 | Actinomycetes and application thereof |
-
2022
- 2022-05-11 CN CN202210515031.5A patent/CN114703111B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103370412A (en) * | 2010-12-04 | 2013-10-23 | 万芬 | RNA stability enhancer |
| CN110218671A (en) * | 2019-05-29 | 2019-09-10 | 江苏师范大学 | One plant of production water streptomycete and its application in biological control |
| CN114231435A (en) * | 2021-10-14 | 2022-03-25 | 商丘师范学院 | Actinomycetes and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114703111A (en) | 2022-07-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112458012A (en) | Bacillus belgii microbial agent and application thereof | |
| CN110257290B (en) | Plant pathogenic bacteria inhibitor, and strain and application thereof | |
| CN109207404B (en) | Siam bacillus YJ15 and application thereof | |
| CN111662837B (en) | Bacillus atrophaeus and application thereof | |
| CN113549578B (en) | Bacillus siamensis BsNlG13 for inhibiting Pyricularia oryzae and promoting seed germination and application thereof | |
| CN113846039A (en) | Bacillus belgii and application thereof | |
| CN112760267A (en) | Lysobacter enzymogenes CX06 for antagonizing xanthomonas campestris and application thereof | |
| CN113980851B (en) | Paracoccus YBH-X with dimethylacetamide degradation capability and application thereof | |
| CN114045238A (en) | Rhodococcus ruber HJM-8 capable of efficiently degrading dimethylacetamide and application thereof | |
| CN113373095A (en) | Application of bacillus cereus FJ2B-137 in biological control of rice sheath blight | |
| CN114790433B (en) | Streptomyces populi ZJPH2021032 and application thereof in preparation of antibacterial agent | |
| CN116426440B (en) | Novel marine sulfur oxidizing bacteria strain and application thereof | |
| CN110295131B (en) | Stenotrophomonas for efficiently degrading polybutylene terephthalate/adipate and application thereof | |
| CN109182216B (en) | Marine streptomyces SCFJ-05 with inhibition effect on succulent plant stem rot | |
| CN108977385B (en) | Streptomyces mismodifier for producing chitinase and application thereof | |
| CN114703111B (en) | Streptomyces hydrogenogenes ZJPH2021031 and application thereof in preparation of antibacterial agent | |
| CN108085275B (en) | Enterobacter ludwigii for inhibiting pathogenic fungi of white muscardine silkworm and application thereof | |
| JP2024037121A (en) | Pbat agricultural film-degrading bacterium and application thereof | |
| CN113234600B (en) | Saline-alkali-tolerant anti-pathogenic fungus aspergillus terreus strain SYAT-1 and application thereof | |
| CN108949622B (en) | Streptomyces for producing chitinase and application thereof | |
| CN110438026B (en) | Bacillus amyloliquefaciens GLD-191 and microbial inoculum, preparation method and application thereof | |
| CN114752534B (en) | Streptomyces that have been described as ZJPH2021033 and use thereof in the preparation of antibacterial agents | |
| CN113980852A (en) | Microbial composition for synergistically degrading benzonitrile herbicide and microbial agent produced by same | |
| CN113373091A (en) | Biocontrol strain bacillus thuringiensis FJ2B-25 for preventing and treating rice sheath blight | |
| CN111849843A (en) | Application of Siamese bacillus B11 in prevention and/or treatment of bamboo shoot dry blight |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |