CN1818056A - Hainan micromonosporin with antibiotic generation - Google Patents
Hainan micromonosporin with antibiotic generation Download PDFInfo
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- CN1818056A CN1818056A CN 200510135592 CN200510135592A CN1818056A CN 1818056 A CN1818056 A CN 1818056A CN 200510135592 CN200510135592 CN 200510135592 CN 200510135592 A CN200510135592 A CN 200510135592A CN 1818056 A CN1818056 A CN 1818056A
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- hainan
- micromonospora
- antibiotic
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- methicillin
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Abstract
The invention opened a Micromonosporaceae which can secrete the antibiotic. The conservation number is CGMCCNo.1551. The antibiotic can inhibit many kinds of the G+ bacteria especially the Staphylococcus aureus and the Mycobacterium tuberculosis. Also it can inhibit the G- bacteria, so it has the good future for the medicine studying.
Description
Technical field
The present invention relates to a strain and produce antibiotic Hainan micromonospora, belong to microbial technology field.
Background technology
Microbiotic is a most widely used medicine in the hospital clinical, to controlling, preventing and treat various infectious diseases to have vital role.Methicillin-resistant staphylococcus aureus (Methicillin-resistant Staphylococcus arueus in recent years, MRSA) separation rate constantly raises, alternative clinically medicine is few simultaneously, MRSA has become " superbacteria " that is difficult to treat, greatly endangering human health, people must constantly seek new medicine source.
Taking up an area of the marine organisms of ball Biological resources 80%, have abundant pathways metabolism, can produce the meta-bolites that a large amount of structures are special, effect is unique, is the important source of novel active.Unique ocean environment makes marine microorganism present higher diversity on species, genomic constitution and ecological functions, is described as the resource " treasure-house " of natural drug, is the new resources of the current natural drug of extremely paying attention to.
Summary of the invention
The technical problem to be solved in the present invention provides a strain can produce antibiotic micromonospora novel species.
For achieving the above object, the present invention is by the following technical solutions:
One strain can produce antibiotic Hainan micromonospora (Micromonospora hainanensis sp.nov.) AM105, and preserving number is CGMCC No.1551.
Hainan micromonospora CGMCC No.1551 is by separating the wild actinomycetes that obtain in the marine bottom sediment of the South Sea, obtaining behind strain improvement.
Hainan micromonospora CGMCC No.1551 has following microbiological property:
(1) form and cultural characters
This bacterial classification can both form well-developed substrate mycelium on multiple substratum, base silk orange (table 1).When cultivating on oat juice substratum, the spore chain of one or more sporulations is arranged, sporophore is different in size, and spore surface is smooth, out-of-shape (Fig. 1 and Fig. 2).
The cultural characteristic of table 1 Hainan micromonospora
| Substratum | Aerial hyphae | Substrate mycelium | But lysochrome |
| GYM agar JCM agar | Do not have | Orange red | Do not have |
| Sauton ' s agar murphy juice agar oat juice agar glycerine asparagine agar Gause I synthetic agar | Few, white does not have few, yellow-white | Orange shallow orange red orange; The light brown Melon yellow of gradual change is orange, and gradual change is light brown | Do not have |
(2) physiological and biochemical property
In 9 kinds of carbon sources commonly used, only utilize glucose, do not utilize L-arabinose, D-wood sugar, D-fructose, L-rhamnosyl, D-N.F,USP MANNITOL, sucrose, raffinose and inositol.Liquefy gelatin does not produce H
2S and class melanochrome, the starch hydrolysis positive can be utilized Mierocrystalline cellulose, the nitrate reduction positive, milk does not solidify, and does not peptonize.
(3) the cell walls chemical composition is analyzed
Full cell hydrolyzed solution contains meso-DAP (meso-diaminopimelic acid, Diaminopimelic acid) and glycine, cell wall II type; Contain wood sugar, pectinose, sugared type D.
(4) Hainan of the present invention micromonospora, its 16S rDNA sequence is as follows:
AGAGTTTGATCATGGCTCAGGACGAACGCTGGCGGCGTGCTTAACACATGCAAGT
CGAGCGGAAAGGCCCTTCGGGGTACTCGAGCGGCGAACGGGTGAGTAACACGTG
AGTAACCTGCCCTAGGCTTTGGGATAACCCTCGGAAACGGGGGCTAATACCGGAT
ATGACCTTCTTCCGCATGGTGGGGGGTGGAAAGTTTTTCGGCCTGGGATGGGCTC
GCGGCCTATCAGCTTGTTGGTGGGGTGATGGCCTACCAAGGCGACGACGGGTAGC
CGGCCTGAGAGGGCGACCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTA
CGGGGGGCAGCAGTGGGGAATATTGCACAATGGGCGGAAGCTGATGCAGCGACG
CCGCGTGAGGGATGACGGCCTTCGGGTTGTAAACCTCTTCCAGCAGGGACGAAG
CGCAAGTGACGGTACCTGCAGAAGAAGCACCGGCCAACTACGTGCCAGCAGCCG
CGGTAAGACGTAGGGTGCGAGCGTTGTCCGGATTTACTGGGCGTAAAGAGCTCGT
AGGCGGCTTGTCGCGTCGACCGTGAAAACTTGGGGCTCAACCCCAAGCCTGCGG
TCGATACGGGCAGGCTAGAGTTCGGTAGGGGAGACTGGAATTCCTGGTGTAGCGG
TGAAATGCGCAGACATCAGGAGGAACACCGGTGGCGAAGGCGGGTCTCTGGGCC
GATACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTG
GTAGTCCACGCTGTAAACGTTGGGCGCTAGGTGTGGGGGGCCTCTCCGGTTTCCT
GTGCCCGCACTAACGCATTAAGCGCCCCGCCTGGGGAGTACGGCCGCAAGGCTAA
AACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGCGGATTAATT
CGATGCAACGCGAAGAACCTTACCTGGGTTTGACATGGCCACAAAACTTCCAGAG
ATGGGAGGTCCTTCGGGGGCGGTCACAGGTGGTGCATGGCTGTCGTCAGCTCGTG
TCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGTTCGATGTTGCC
AGCGCGTTATGGCGGGGACTCATCGAAGACTGCCGGGGTCAACTCGGAGGAAGG
TGGGGATGACGTCAAGTCATCATGCCCCTTATGTCCAGGGCTTCACGCATGCTACA
ATGGCCGGTACAATGGGTTGCGATGCCGTGAGGTGGAGCGAATCCCAAAAAGCC
GGTCTCAGTTCGGATCGGGGTCTGCAACTCGACCCCGTGAAGTCGGAGTCGCTAG
TAATCGCAGATCAGCAACGCTGCGGTGAATACGTTCCCGGGCCTTGTACACACCG
CCCGTCACGTCACGAAAGTCGGCAACACCCGAAGCCGGTGGCCCAACCCTTGTG
GAGGGAGCCGTCGAAGGTGGGGCTGGCGATTGGGACGAAGTCGTAACAAGGTAA
CC。
Hainan micromonospora CGMCC No.1551 that the present invention relates to, identify through multinomial classification, be defined as a novel species of micromonospora, called after Hainan micromonospora (Micromonospora hainanensis sp.nov), carried out culture presevation on December 5th, 2005 in China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC), and the proof survival, its preservation registration number is CGMCC No.1551.The preservation address is the No. 13, North No.1 Row, Zhongguancun, Haidian District, Beijing City, Institute of Microorganism, Academia Sinica.
Advantage of the present invention is: the microbiotic that this Hainan micromonospora produces is to multiple G
+, G
-Bacterium has stronger restraining effect, especially methicillin-resistant staphylococcus aureus and Mycobacterium tuberculosis is had obvious restraining effect.
The invention will be further described below in conjunction with specification drawings and specific embodiments.
Description of drawings
Fig. 1 is the mycelia and the fibrillae of spores form of bacterial strain of the present invention, 1500 *.
Fig. 2 is the mycelia and the fibrillae of spores form of bacterial strain of the present invention, 10000 *.
Embodiment
The invention will be further described below in conjunction with specific embodiment:
Embodiment 1: the seed selection of Hainan micromonospora CGMCC No.1551
With Hainan micromonospora of natural separation, obtain the big and full bacterial strain of a strain bacterium colony by natural seed selection, color is orange.With this bacterial strain is starting strain, through ultraviolet mutagenesis 10～60s, has obtained mutagenesis effect preferably, obtains a strain CGMCC No.1551, and its fermented liquid is increased to 40mm to the antibacterial circle diameter of MRSA by 19mm, is 2.1 times of starting strain.
Embodiment 2: the cultivation of Hainan micromonospora CGMCC No.1551
Culture condition:
(1) slant culture: cultivated 6 days.Substratum: yeast powder O.4%, malt meal 1%, glucose 0.4%, agar 2%, pH7.2,28 ℃ of culture temperature.
(2) seed culture: insert the seed liquor 28 ℃ of 200rpm shake-flask culture 48 hours from the activated single bacterium colony of inclined-plane picking.Substratum: glucose 1%, soyflour 1%, yeast extract paste 1%, Zulkovsky starch O.5%, KH
2PO
40.05%, natural old seawater preparation, pH7.2.
(3) fermentation culture: the inoculum size by 10% inserts seed culture medium, shake-flask culture 6 days.Substratum: glucose 0.5%, starch 1.5%, soyflour 1%, yeast extract paste 0.5%, K
2HPO
40.025%, with 50% natural old seawater (fresh seawater leaves standstill all backs) and the preparation of 50% distilled water, 7.2,28 ℃ of initial pH value, shaking speed 200r/min.
Embodiment 3: the cultivation of Hainan micromonospora CGMCC No.1551
Activatory CGMCC No.1551 placed shake bottle and carry out seed culture, corresponding substratum is with embodiment 2, and condition is 30 ℃ of temperature, and shaking speed 180rpm cultivates 45h.Inoculum size with 8% inserts fermention medium, and substratum uses natural full sea water preparation, and 28 ℃ of 200rpm cultivated 7 days.
The checking of embodiment 4. pharmacodynamics
One. antibiotic acquisition:
Method by embodiment 3 is carried out fermentation culture, and fermented liquid carries out following processing:
1. be collected in the tunning that stopped fermentation on the 6th day, the centrifugal 10min of 8000r/min collects supernatant liquor.
2. adopt an amount of ethyl acetate that fermented supernatant fluid is carried out 4～5 extractions, the combining extraction liquid concentrating under reduced pressure gets the microbiotic crude product.
3. employing silica gel column chromatography, eluent is: sherwood oil: ethyl acetate 6: 1 → 3: 1 → 1: 1 → 100% ethyl acetate gradient elution, fraction collection elutriant.
4. concentrating under reduced pressure is collected active peak, through Sephadex LH20 column chromatography, obtains purer microbiotic crude product.
Two. antibacterial tests
The microbiotic sample that employing plate doubling dilution mensuration obtains the results are shown in Table 2 to the minimum inhibition concentration MIC of a plurality of detection bacterial strains.As can be seen, the microbiotic that this Hainan micromonospora produces has stronger restraining effect to multiple G+ bacterium, especially methicillin-resistant staphylococcus aureus and Mycobacterium tuberculosis is had obvious restraining effect, and also there is certain inhibition in the G-bacterium.
Table 2 Hainan micromonospora produces antibiotic bacteriostatic activity and measures
| Strains tested | AM105-II(μg/mL) |
| MRSA1 MRSA2 MRSA3 MRSA4 MRSA5 MRSA6 MRSA7 MRSA8 Bacillus subtili Sarcina lutea E.coli Pseudomonus aeruginosa Mycobacterium tuberculosis Candida albicans Aspergillus niger | ≤0.03 0.06 ≤0.03 0.125 8 0.25 0.125 0.125 ≤0.03 ≤0.03 32 64 0.06 - - |
Annotate: MRSA1: methicillin-resistant, erythromycin, Ciprofloxacin, amikacin; MRSA2: methicillin-resistant, erythromycin, tsiklomitsin; MRSA3: methicillin-resistant, tsiklomitsin, erythromycin, Ciprofloxacin, amikacin; MRSA4: methicillin-resistant, Ciprofloxacin, amikacin, erythromycin; MRSA5: methicillin-resistant, tsiklomitsin, erythromycin, Ciprofloxacin; MRSA6: methicillin-resistant, erythromycin, tsiklomitsin; MRSA7: methicillin-resistant, erythromycin, tsiklomitsin; MRSA8: methicillin-resistant, tsiklomitsin, erythromycin, Ciprofloxacin, amikacin.
Claims (1)
1. a strain produces antibiotic Hainan micromonospora, and preserving number is CGMCC No.1551.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005101355929A CN100340660C (en) | 2005-12-30 | 2005-12-30 | Hainan micromonosporin with antibiotic generation |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005101355929A CN100340660C (en) | 2005-12-30 | 2005-12-30 | Hainan micromonosporin with antibiotic generation |
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|---|---|
| CN1818056A true CN1818056A (en) | 2006-08-16 |
| CN100340660C CN100340660C (en) | 2007-10-03 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102978133A (en) * | 2012-11-16 | 2013-03-20 | 中国科学院南海海洋研究所 | Micromonospora Rosaria and method for preparing a plurality of antibiotics by Micromonospora Rosaria |
-
2005
- 2005-12-30 CN CNB2005101355929A patent/CN100340660C/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102978133A (en) * | 2012-11-16 | 2013-03-20 | 中国科学院南海海洋研究所 | Micromonospora Rosaria and method for preparing a plurality of antibiotics by Micromonospora Rosaria |
| CN102978133B (en) * | 2012-11-16 | 2014-08-06 | 中国科学院南海海洋研究所 | Micromonospora Rosaria and method for preparing a plurality of antibiotics by Micromonospora Rosaria |
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|---|---|
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