CN108130292B

CN108130292B - Streptomyces marinus S063 and application of anticomplement activity thereof

Info

Publication number: CN108130292B
Application number: CN201810010543.XA
Authority: CN
Inventors: 赵心清
Original assignee: Shanghai Jiaotong University
Current assignee: Shanghai Jiaotong University
Priority date: 2018-01-04
Filing date: 2018-01-04
Publication date: 2021-03-12
Anticipated expiration: 2038-01-04
Also published as: CN108130292A

Abstract

The invention discloses a strain of marine streptomyces S063 and its application in anticomplement activity; the marine Streptomyces is marine Streptomyces (Streptomyces sp.) S063CGMCC No. 14582. The fermentation liquor of the marine streptomyces has obvious anti-complement activity as shown by a complement hemolysis test. The invention also discloses a fermentation condition for preparing the marine streptomyces anticomplementary active substance.

Description

Streptomyces marinus S063 and application of anticomplement activity thereof

Technical Field

The invention relates to the technical field of microorganisms, in particular to a marine streptomyces S063 strain and application of the marine streptomyces S063 strain in anticomplement activity.

Background

The marine environment occupies 70.8 percent of the total area of the earth, contains rich microbial resources, and the unique environmental characteristics of the sea, such as low temperature, low nutrition, high pressure, high salinity and the like, create the special metabolic mode and metabolic products of the marine microorganisms, and the actinomycetes have the capability of synthesizing various secondary metabolic products due to the complex morphological differentiation process. In recent years, with the development of molecular biology techniques, more and more marine actinomycetes have been discovered, and many secondary metabolites derived from marine actinomycetes have been identified as having various bioactive functions such as antitumor, antibacterial, antimalarial and insecticidal functions, and enzyme inhibitors (Zotchev, J Biotechnol,2012,158: 168-75). The active substances with novel structure and multiple functions generated by the marine actinomycetes become important sources for the development of natural active products and new drugs.

Complement (Complement) is a group of activated enzymatically active proteins present in normal human and animal serum and tissue fluids, which can aid and Complement specific antibodies, mediate immunolysis, hemolysis, and is known as Complement (Min, et al. biol Pharm Bull,2003,26: 1042-4). The complement system is one of the important immune defense systems of the human body. Normal activation of the complement system plays an important role in the destruction of foreign microorganisms, the removal of damaged or dead cells and tissues in the body, and the maintenance of the balance of the body (Hourcade, et al. molecular immunology,2016,69: 62-9). The complement activation pathways are mainly three, the Classical pathway (classic pathway), Alternative pathway (Alternative pathway), and lectin (MBL) pathway (Terminal pathway). However, abnormal complement activation can cause many diseases such as rheumatoid arthritis, senile dementia, systemic lupus erythematosus, ischemic reperfusion, acute myocardial infarction, acute respiratory distress syndrome, and the like (Qu, et al. mol. Immunol,2009,47: 185-95). In addition, excessive complement activation can also cause substantial damage to tissues such as blood vessels (Alper, et al. Science,1976,191:1275-6), kidneys (Cooper, J Exp Med,1973,137:451-60), joints (Holden, Science,1980,207:161), and red blood cells (Caldwell, Rheum Dis Clin North Am,1999,25: 919-28). Studies have also found that severe atypical pneumonia (SARS) is also associated with over-activation of the complement system (Peiris, et al. Lancet,2003,361: 1767-72; Wong, et al. BMJ,2003,326: 1358-62). Therefore, the development of drugs having anti-complement activity has attracted much attention.

The chemosynthetic anticomplementary inhibitors have the disadvantages of high cost, poor selectivity, reduction of human body defense functions after long-term use, various side effects and the like, and the anticomplementary active ingredients in natural product sources have low development cost and can be directly digested and absorbed in vivo (thank you, etc., Chinese medicine guidance, 2013,33: 28-31). Therefore, natural product sources of anti-complement active substances have attracted widespread attention from scholars at home and abroad. In recent years, anticomplementary active substances derived from natural products are continuously discovered, and show good application prospects of anticomplementary activity. To date, many substances with anticomplementary activity have been isolated from animals, plants and microorganisms, including polysaccharides, flavones, steroids, saponins, terpenes, alkaloids, etc. (Xuxiana et al, natural products research and development, 2015,02:355-359+ 327).

Compared with the limitations of long growth period, low yield, difficult quality control and the like of plant resources, the microorganism has unique advantages, such as fast growth and reproduction, easy culture, controllable growth process, high yield, suitability for industrial production and various secondary metabolites of the microorganism. In addition, microorganisms are easy to genetically manipulate, a large number of analogs can be obtained by combinatorial biosynthesis and the like, and the study of structure and drug effect and improvement of activity are easier. Therefore, the development of the anticomplementary active substance derived from the microorganism has important significance for protecting limited plant resources and species diversity, and has huge application and development prospects in the aspect of mass production.

At present, the research on the anticomplementary active substances from natural products is still in the primary stage, and a plurality of problems are still to be solved, such as the unclear structure, the unknown action targets and the unknown toxic and side effects of the anticomplementary active ingredients separated from a plurality of natural products, and the like. The current clinical need for new complement inhibitors is increasing, and there is a need to enhance the basic research on anticomplementary active ingredients in natural products, particularly the development of anticomplementary active ingredients of microbial origin.

Disclosure of Invention

The invention aims to provide a strain of marine streptomyces S063 and the application of anticomplement activity thereof; the metabolite of the marine actinomycete streptomycete (Streptomyces sp) S063 has obvious effect of inhibiting complement activity.

The marine actinomycete streptomycete (Streptomyces sp.) S063 is separated from a sea mud sample of Liaoning large and small islands, and a purification culture medium is a Bennett (Bennett) culture medium which grows well on the Bennett culture medium; the colony of the marine streptomyces S063 is convex, the surface is wrinkled, and offwhite spores are contained.

The marine Streptomyces (Streptomyces sp.) S063 has a strain preservation number of CGMCC No.14582, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, and has a preservation unit address of No. 3 Xilu-Beichen No.1 of Beijing city and Inward area, a microbial research institute of Chinese academy of sciences, a postal code of 100101, and a preservation date of 2017, 8 months and 30 days.

The purpose of the invention is realized by the following technical scheme:

in a first aspect, the invention relates to a strain of marine Streptomyces, which is S063CGMCC No. 14582.

Preferably, the purified culture medium of the marine streptomyces is a bennett culture medium; the composition of each 1L of the bennett medium was: 10g of glucose, 2g of peptone, 1g of yeast powder, 1g of beef extract, 15g of agar and distilled water, wherein the volume is constant to 1L, the pH value is 7.2, and the wet heat sterilization is carried out at 121 ℃ for 20 min.

Preferably, the 16S rRNA nucleotide complete sequence of the marine streptomyces S063 is shown as SEQ ID NO.1, and the total length is 1429 bp.

Preferably, the metabolite of Streptomyces marinus has anti-complement activity.

In a second aspect, the invention relates to the use of the marine streptomyces in the preparation of a drug with anti-complement activity.

Preferably, the metabolic product of the marine streptomyces is used for preparing the anti-complement active medicine.

Preferably, the metabolic product of the marine streptomyces is prepared by a method comprising the following steps:

s1, fermenting the marine streptomyces S063;

s2, chopping an agar plate of a marine streptomyces S063 solid culture medium, adding methanol with the same amount, soaking for 24h, and concentrating the obtained extract; dissolving with water with the volume of 1/10, and filtering insoluble precipitate to obtain the metabolic product of the marine streptomycete;

or, the supernatant fluid after the S063 liquid fermentation liquid is centrifuged is extracted by ethyl acetate, and mycelium is extracted by methanol; respectively concentrating the obtained extract; dissolving with water with the volume of 1/10, and filtering insoluble precipitate to obtain the metabolic product of the marine streptomycete.

Preferably, in step S1, the fermentation uses a solid medium or a liquid medium; the solid culture medium comprises soybean tryptone, No. two, No. three, No. eight, No. seventeen and No. nineteen culture media, and the liquid culture medium comprises a liquid fermentation culture medium of soybean tryptone;

every 1L of the soybean tryptone culture medium comprises: 17g of tryptone, 3g of soybean peptone, 5g of sodium chloride, 2.5g of glucose, 2.5g of dipotassium hydrogen phosphate and distilled water, wherein the volume is constant to 1L, and the wet heat sterilization is carried out at 121 ℃ for 20 min;

the composition of the second solid culture medium is as follows for every 1L: 20g of soluble starch, 10g of glucose, 5g of peptone, 5g of yeast powder, 5g of calcium carbonate, 15g of agar, and distilled water with constant volume of 1L, and carrying out moist heat sterilization at 121 ℃ for 20 min;

each 1L of the No. three solid culture medium consists of: 20g of soluble starch, 25g of soybean meal, 2g of ammonium sulfate, 2g of sodium chloride, 0.5g of dipotassium phosphate, 5.0g of calcium carbonate and 15g of agar, wherein the pH value is 7.2, the volume of distilled water is constant to 1L, and the wet heat sterilization is carried out for 20min at 121 ℃;

the composition of the No. eight solid culture medium is 1L: 40g of malt extract, 2g of glucose, 1g of ammonium hydrogen phosphate, 15g of agar and distilled water to a constant volume of 1L, and carrying out moist heat sterilization at 121 ℃ for 20 min;

every 1L of the No. seventeen solid culture medium comprises: 10g of glucose, 10g of soybean meal, 10g of corn steep liquor, 5g of glycerol, 5g of yeast powder, 5g of sodium chloride, 2.0g of calcium carbonate and 15g of agar, wherein the pH value is 5.7, the volume of distilled water is constant to 1L, and the wet heat sterilization is carried out for 20min at 121 ℃;

the composition of nineteen solid culture media is as follows for each 1L: 20g of soluble starch, 5g of cottonseed powder, 2.5g of yeast powder, 1g of sodium chloride, 0.75g of dipotassium phosphate, 1g of magnesium sulfate heptahydrate, 3g of calcium carbonate, 15g of agar and distilled water, wherein the volume is fixed to 1L, and the wet heat sterilization is carried out for 20min at 121 ℃.

Preferably, step S1 specifically includes:

A. preparation of marine streptomyces S063 seed liquid: inoculating hypha and spore on the flat plate into a soybean tryptone culture medium, and culturing at 30 ℃ and 200rpm for 48h to obtain a seed solution;

B. fermentation of marine streptomyces S063: inoculating every 0.1mL of the seed solution into 15mL of the solid medium agar plate, and culturing for 7 days at the temperature of 30 ℃.

Or, marine streptomyces S063 liquid fermentation: inoculating every 1mL of the seed solution into 50mL of the fermentation medium, and culturing for 7 days at the temperature of 30 ℃.

Preferably, in step S2, the fermentation medium agar plate is cut, and an equal amount of methanol is added to soak the cut plate for 24 hours; extracting supernatant after the fermentation liquor is centrifuged by ethyl acetate for 3 times, and performing isovolumetric extraction; the mycelium was extracted 3 times with methanol and the same volume.

The invention has the following beneficial effects: the invention provides a strain of marine streptomyces, and complement hemolysis test shows that fermentation broth of the strain has obvious inhibition effect on complement and can protect blood cells. The invention also discloses a metabolite of the marine streptomyces with anticomplementary effect, and the preparation method simply prepares the metabolite with anticomplementary activity by controlling the solid fermentation condition of the marine streptomyces S063.

Drawings

FIG. 1 shows the colony morphology of strain S063;

FIG. 2 shows the anti-complement activity of methanol extracts of different solid media of strain S063; wherein, 2, 3, 8, 17 and 19 are 5 kinds of culture media for optimization, and the methanol extract of the No. 2, 8 and 19 culture media has better activity; s is standard hemolysis and M is mechanical hemolysis.

Detailed Description

The present invention will be further described with reference to the following examples.

Example 1 identification of the Strain S063

The streptomyces is identified as streptomyces by combining morphological characteristics, physiological and biochemical characteristics, 16S rRNA sequences and the like of the streptomyces. The specific identification results are as follows:

1. form of cell

Strain S063 grew well on bennett medium with convex colonies, surface wrinkles, white spores (see figure 1).

2. Physiological and biochemical characteristics

Streptomyces S063 can grow on soybean tryptone medium with additional salt concentration of 0-12%, grows well and produces spores at 0-9%, and when 12%, the colonies are very small and grow slowly.

TABLE 1 salt tolerance of Actinomycetes S063

NaCl content (%)	0	3	6	9	12
						Growth conditions	++	+++	++	++	+

Note: the greater the "+" number, the better the growth

3.16S rRNA identification

According to the comparison analysis of phylogenetic trees, the strain S063 and Streptomyces setonii NBRC 13085^T(99.44%) showed the highest sequence homology.

The total length of the 16S rRNA nucleotide complete sequence of the streptomyces S063 is 1429bp, and the sequence is shown as follows (SEQ ID NO. 1):

ACCGTGCGGCATGCTACCATGCAGTCGAACGATGAAGCCGCTTCGGTGGTGGATTAGTGGCGAACGGGTGAGTAACACGTGGGCAATCTGCCCTTCACTCTGGGACAAGCCCTGGAAACGGGGTCTAATACCGGATAACACTCTGTCCCGCATGGGACGGGGTTGAAAGCTCCGGCGGTGAAGGATGAGCCCGCGGCCTATCAGCTTGTTGGTGGGGTAATGGCCTACCAAGGCGACGACGGGTAGCCGGCCTGAGAGGGCGACCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGAAAGCCTGATGCAGCGACGCCGCGTGAGGGATGACGGCCTTCGGGTTGTAAACCTCTTTCAGCAGGGAAGAAGCGAAAGTGACGGTACCTGCAGAAGAAGCGCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGCGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGAGCTCGTAGGCGGCTTGTCACGTCGGATGTGAAAGCCCGGGGCTTAACCCCGGGTCTGCATTCGATACGGGCTAGCTAGAGTGTGGTAGGGGAGATCGGAATTCCTGGTGTAGCGGTGAAATGCGCAGATATCAGGAGGAACACCGGTGGCGAAGGCGGATCTCTGGGCCATTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGTTGGGAACTAGGTGTTGGCGACATTCCACGTCGTCGGTGCCGCAGCTAACGCATTAAGTTCCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCAGCGGAGCATGTGGCTTAATTCGACGCAACGCGAAGAACCTTACCAAGGCTTGACATATACCGGAAAGCATCAGAGATGGTGCCCCCCTTGTGGTCGGTATACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTTCTGTGTTGCCAGCATGCCCTTCGGGGTGATGGGGACTCACAGGAGACTGCCGGGGTCAACTCGGAGGAAGGTGGGGACGACGTCAAGTCATCATGCCCCTTATGTCTTGGGCTGCACACGTGCTACAATGGCCGGTACAATGAGCTGCGATGCCGCGAGGCGGAGCGAATCTCAAAAAGCCGGTCTCAGTTCGGATTGGGGTCTGCAACTCGACCCCATGAAGTCGGAGTTGCTAGTAATCGCAGATCAGCATTGCTGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACGTCACGAAAGTCGGTAACACCCGAAGCCGGTGGCCCAACCCCTTGTGGGAGGGAGCTTCGAAGGTGATGGGGCC。

example 2 preparation of metabolite of Streptomyces marinus S063 having anticomplementary Effect

(1) Fermentation of Streptomyces S063

C. Preparation of marine streptomyces S063 seed liquid: inoculating hypha and spore on the flat plate into a soybean tryptone culture medium, and culturing at 30 ℃ and 200rpm for 48h to obtain a seed solution;

D. fermentation of marine streptomyces S063: inoculating the seed solution into 15mL of solid medium agar plates with the inoculation amount of 0.1mL, including No. two, No. three, No. eight, No. seventeen and No. nineteen culture media, and culturing for 7 days at the temperature of 30 ℃.

E. Marine streptomyces S063 liquid fermentation culture: inoculating every 1mL of the seed solution into 50mL of soybean tryptone fermentation medium, and culturing for 7 days at the temperature of 30 ℃.

The soybean tryptone culture medium (1#) consists of: 17g of tryptone, 3g of soybean peptone, 5g of sodium chloride, 2.5g of glucose, 2.5g of dipotassium hydrogen phosphate and distilled water, wherein the volume is constant to 1L, and the wet heat sterilization is carried out at 121 ℃ for 20 min;

the second solid culture medium (2#) consists of: 20g of soluble starch, 10g of glucose, 5g of peptone, 5g of yeast powder, 5g of calcium carbonate, 15g of agar, and distilled water with constant volume of 1L, and carrying out moist heat sterilization at 121 ℃ for 20 min;

1L of the No. three solid culture medium (3#) comprises: 20g of soluble starch, 25g of soybean meal, 2g of ammonium sulfate, 2g of sodium chloride, 0.5g of dipotassium phosphate, 5.0g of calcium carbonate and 15g of agar, wherein the pH value is 7.2, the volume of distilled water is constant to 1L, and the wet heat sterilization is carried out for 20min at 121 ℃;

the No. eight solid culture medium (8#) consists of: 40g of malt extract, 2g of glucose, 1g of ammonium hydrogen phosphate, 15g of agar and distilled water to a constant volume of 1L, and carrying out moist heat sterilization at 121 ℃ for 20 min;

1L of the No. seventeen solid culture medium (17#) comprises: 10g of glucose, 10g of soybean meal, 10g of corn steep liquor, 5g of glycerol, 5g of yeast powder, 5g of sodium chloride, 2.0g of calcium carbonate and 15g of agar, wherein the pH value is 5.7, the volume of distilled water is constant to 1L, and the wet heat sterilization is carried out for 20min at 121 ℃;

the nineteen solid culture media (19#) consist of: 20g of soluble starch, 5g of cottonseed powder, 2.5g of yeast powder, 1g of sodium chloride, 0.75g of dipotassium phosphate, 1g of magnesium sulfate heptahydrate, 3g of calcium carbonate, 15g of agar and distilled water, wherein the volume is fixed to 1L, and the wet heat sterilization is carried out for 20min at 121 ℃.

(2) Preparation of metabolite of Streptomyces S063 having anticomplementary (complement inhibition) Activity

Cutting agar plate of Streptomyces S063 fermentation medium, adding equivalent methanol, soaking for 24h, and concentrating the obtained extract with thin film rotary evaporator (35 deg.C); dissolving with 1/10 vol water, and filtering to remove insoluble precipitate; s063 liquid fermentation liquid is centrifuged for 15min at 5000rpm, supernatant is extracted by ethyl acetate (3 times of extraction and equal volume of extraction), mycelia is extracted by methanol (3 times of extraction and equal volume of extraction), obtained extract liquid is respectively concentrated by a thin film rotary evaporator (35 ℃), and then a sample is respectively dissolved by water with 1/10 volumes, insoluble precipitates are filtered out, and the metabolite of the streptomyces S063 with obvious anticomplementary activity is obtained.

(3) Hemolytic assay for anticomplementary Activity

Diluting 5 × BBS buffer to 1 × BBS buffer; centrifuging sheep red blood cells at 5000rpm for 5min, discarding the supernatant, washing the sheep red blood cells twice with BBS buffer solution, and diluting the sheep red blood cells to 2%; diluting 1:4000 hemolysin to 1:1000 by BBS buffer solution; diluting 1:20 complement reagent with BBS buffer solution to critical concentration of 1: 160; mixing 100 μ L complement and 100 μ L sample, adding 100 μ L complement into test tube, adding BBS, and water-bathing at 37 deg.C for 10min (the sample number of water-bathing is shown in Table 2); adding hemolysin and sheep red blood cells into all sample tubes respectively, supplementing 0.6mL by BBS buffer solution, and preparing a hemolysis group; placing into 37 deg.C water bath for 30min, taking outCooling on ice; centrifuging at 4 deg.C and 5000rpm for 10min, and measuring OD with 0.2mL₄₀₅And (4) light absorption value.

After testing various culture media, as shown in figure 2 and table 3, when S063 is cultured in solid culture medium No. 2, 8, 19, the culture extract has obvious anticomplementary effect and can protect cells from being damaged by a complement system. S063 also has good anticomplementary activity in soybean trypsin liquid medium, and the anticomplementary activity of ethyl acetate extract of supernatant is 43.1%, and the anticomplementary activity of mycelium methanol extract is 42.4%.

Sample M is a mechanical hemolysis control, no complement system, and therefore no hemolysis; the sample S is a full-hemolytic control, the sheep-Yang red blood cells in the sample S are combined with hemolysin to form sensitized sheep-Yang red blood cells, the sensitized sheep-Yang red blood cells are combined with complement again to activate the sensitized sheep-Yang red blood cells, and the complement has cytolytic effect to cause red blood cell swelling and hemolysis.

Barbital Buffer Solution (BBS) described above: taking 2.875g of barbital sodium, dissolving in 250mL of hot and air water, and adding 42.5g of NaCl and MgCl₂·6H₂O 0.84g、CaCl₂0.14g of barbital and 1.0g of barbital, and 1000mL of triple distilled water were added to prepare a 5-fold concentrated barbital buffer (5 XBBS). When in use, 5 XBBS is diluted into Barbital Buffer Solution (BBS) by using triple-distilled water.

The above 2% Sheep Red Blood Cell (SRBC): 2mL of sheep blood is taken, washed by BBS, centrifuged for 3min at 800rpm, repeatedly washed for 3 times, added with BBS to 10mL, mixed evenly, added with about 0.5mL of erythrocyte hematocrit tube, centrifuged for 10min at 3000rpm, the volume percentage of erythrocytes is measured, and proper amount of BBS is added proportionally to prepare 2% SRBC.

1:1000 hemolysin: taking a proper amount of hemolysin, adding BBS, and preparing into a 1:1000 hemolysin solution.

TABLE 2 anticomplementary Activity test reagent ratios

Sample number	Sample name	Serum	Hemolysin	Red blood cell	Buffer solution	Water (W)	Sample (I)
								1	Standard of merit	0.1	0.1	0.1	0.3
2*	Background of serum	0.1			0.5
								3*	Background of hemolysin		0.1		0.5
4	Total hemolysis			0.1		0.5
								5	Mechanical hemolysis			0.1	0.5
6	Haemolysis of serum	0.1		0.1	0.4
								7	Hemolysin hemolysis		0.1	0.1	0.4
8*	Sample background				0.5		0.1
								9	Hemolysis of a sample			0.1	0.4	0.1
10	Sample (I)	0.1	0.1	0.1	0.2		0.1

Remarking: the samples are prepared according to the following mixture ratio, the mixture ratio can be measured without the mixture ratio, and the mixture ratio is buffer solution, the samples, serum, hemolysin and red blood cells in sequence

The calculation formula for the detection of anti-complement activity is as follows:

standard hemolysis-serum hemolysis-hemolysin hemolysis + mechanical hemolysis

Sample hemolysis-serum hemolysis-hemolysin hemolysis-sample hemolysis +2 × mechanical hemolysis

Anticomplementary activity (%). cndot.1-sample hemolysis/standard hemolysis). times.100

Table 3, S063 classical pathway anticomplementary Activity in different solid media (%)

	2	3	8	17	19
						M	59.30	2.79	95.33	74.36	48.24
E	0	1.76	0	0.82	1.48

Note: m: extracting the solid culture with methanol; e: the solid culture was extracted with ethyl acetate.

Sequence listing

<110> Shanghai university of transportation

<120> marine streptomyces S063 and its anticomplement activity application

<130> 2017

<141> 2018-01-04

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1429

<212> DNA

<213> Streptomyces sp. S063

<400> 1

accgtgcggc atgctaccat gcagtcgaac gatgaagccg cttcggtggt ggattagtgg 60

cgaacgggtg agtaacacgt gggcaatctg cccttcactc tgggacaagc cctggaaacg 120

gggtctaata ccggataaca ctctgtcccg catgggacgg ggttgaaagc tccggcggtg 180

aaggatgagc ccgcggccta tcagcttgtt ggtggggtaa tggcctacca aggcgacgac 240

gggtagccgg cctgagaggg cgaccggcca cactgggact gagacacggc ccagactcct 300

acgggaggca gcagtgggga atattgcaca atgggcgaaa gcctgatgca gcgacgccgc 360

gtgagggatg acggccttcg ggttgtaaac ctctttcagc agggaagaag cgaaagtgac 420

ggtacctgca gaagaagcgc cggctaacta cgtgccagca gccgcggtaa tacgtagggc 480

gcaagcgttg tccggaatta ttgggcgtaa agagctcgta ggcggcttgt cacgtcggat 540

gtgaaagccc ggggcttaac cccgggtctg cattcgatac gggctagcta gagtgtggta 600

ggggagatcg gaattcctgg tgtagcggtg aaatgcgcag atatcaggag gaacaccggt 660

ggcgaaggcg gatctctggg ccattactga cgctgaggag cgaaagcgtg gggagcgaac 720

aggattagat accctggtag tccacgccgt aaacgttggg aactaggtgt tggcgacatt 780

ccacgtcgtc ggtgccgcag ctaacgcatt aagttccccg cctggggagt acggccgcaa 840

ggctaaaact caaaggaatt gacgggggcc cgcacaagca gcggagcatg tggcttaatt 900

cgacgcaacg cgaagaacct taccaaggct tgacatatac cggaaagcat cagagatggt 960

gccccccttg tggtcggtat acaggtggtg catggctgtc gtcagctcgt gtcgtgagat 1020

gttgggttaa gtcccgcaac gagcgcaacc cttgttctgt gttgccagca tgcccttcgg 1080

ggtgatgggg actcacagga gactgccggg gtcaactcgg aggaaggtgg ggacgacgtc 1140

aagtcatcat gccccttatg tcttgggctg cacacgtgct acaatggccg gtacaatgag 1200

ctgcgatgcc gcgaggcgga gcgaatctca aaaagccggt ctcagttcgg attggggtct 1260

gcaactcgac cccatgaagt cggagttgct agtaatcgca gatcagcatt gctgcggtga 1320

atacgttccc gggccttgta cacaccgccc gtcacgtcac gaaagtcggt aacacccgaa 1380

gccggtggcc caaccccttg tgggagggag cttcgaaggt gatggggcc 1429

Claims

1. A strain of marine Streptomyces is characterized in that the marine Streptomyces is marine Streptomyces (Streptomyces sp.) S063, and the preservation number of the marine Streptomyces is CGMCC No. 14582.

2. Use of the marine streptomyces of claim 1 in the preparation of a medicament with anti-complement activity.

3. Use according to claim 2, wherein the metabolite of Streptomyces marinus is used for the preparation of an anticomplementary active drug.

4. The use according to claim 3, wherein the metabolite of Streptomyces marinus is produced by a method comprising the steps of:

s1, fermenting the marine streptomyces S063;

5. The use of claim 3, wherein in step S1, the fermentation uses a solid medium or a liquid medium; the solid culture medium comprises soybean tryptone, No. two, No. three, No. eight, No. seventeen or No. nineteen culture media, and the liquid culture medium comprises a liquid fermentation culture medium of soybean tryptone;

6. The use according to claim 5, wherein step S1 specifically comprises:

B. fermentation of marine streptomyces S063: inoculating every 0.1mL of the seed solution into 15mL of the solid medium agar plate, and culturing for 7 days at the temperature of 30 ℃;

7. The method for preparing the metabolite of marine streptomyces DUT11 as claimed in claim 6, wherein in step S2, the fermentation medium agar plate is cut up, added with equal amount of methanol, and soaked for 24 h; extracting supernatant after the fermentation liquor is centrifuged by ethyl acetate for 3 times, and performing isovolumetric extraction; the mycelium was extracted 3 times with methanol and the same volume.