CN103145716A - Technique for preparing and separating cyclic dipeptide C6 from phellinus igniarius bacteria in high pressure opposite-phase mode - Google Patents
Technique for preparing and separating cyclic dipeptide C6 from phellinus igniarius bacteria in high pressure opposite-phase mode Download PDFInfo
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Abstract
The invention discloses a method for separating cyclic dipeptide C6 from phellinus igniarius bacteria (Phellinusigniarius (LexFr) Quel, phellinuslinteus (BerketCurt) Teng and Phellinushartigii (AlleschetSchnabl) Imaz). Crude extracts are prepared from the phellinus igniarius bacteria at first, normal phase silica gel chromatography is carried out, chloroform and carbinol are adopted for elution, eluant liquid is appropriately merged, and drying is carried out under reduced pressure to obtain the cyclic dipeptide C6, namely hexahydrogen-7-hydroxy-3-(2-methyl propyl group) pyrrole [1,2-a] pyrazine-1,4-diketone.
Description
Technical field
The invention belongs to the biotech medicine product field.
Background technology
Phellinus, the sporophore stockless, the flat semisphere of cap or the shape of a hoof, 2-12*3-21 centimetre, thick 1.5-10 centimetre, wooden, shallow liver brown to lead or black, often be full of cracks always, without cot, there is trickle fine hair at initial stage, rear change is without hair, the concentric ring rib is arranged. the edge is blunt, dark cinnamon is to light coffee color, downside is without thalamium. the bacterial context dark brown, firmly, wooden. tube and bacterial context are closely homochromy, multilayer, but level is not obvious, old tube layer is full of white hypha. and mouth of pipe rust brown is to dark reddish brown, circular, every millimeter 4-5. spore is subsphaeroidal, smooth, colourless, the 5-6*3-4 micron. the bristle top is sharp-pointed, base portion expands, the 10-25*5-7 micron. mycelia is branch not, without tabula, diameter 3-5 micron.
At present, the purification process of epiphyte product is more, mainly contain precipitation classification, preparation property high performance liquid chromatography, column chromatography, ultrafiltration process, centrifugation chromatography, etc. several method.And wherein use maximum column chromatographies that surely belongs to, and be divided into two classes: the one, only have the gel filtration chromatography of molecular sieve effect, gel commonly used has dextrane gel and sepharose.The 2nd, ion exchange chromatography.But, mainly for separating of polysaccharide, hyaluronic acid etc., the product that obtains after most the separation is mixture.The present invention uses multiple chromatographic technique to combine, and resolution is high, and using this invention can be exquisite to sterling.
Summary of the invention
The invention discloses a kind of Phellinus bacterium (phelliuns igniarius
Phellinus igniarius(L ex Fr) Quel, phellinus linteus
Phellinus linteus(Berk et Curt) Teng, Phellinus baumii, Kazakhstan base of a fruit phellinus
Phellinus hartigii(Allesch et Schnabl) Imaz) separation method of ring dipeptides C6 in.At first prepare Phellinus bacterium crude extract, then carry out the purification on normal-phase silica gel chromatography, adopt chloroform and methanol-eluted fractions;, be then the methanol gel chromatography, after HPLC detects; carry out the anti-phase preparation of high pressure; detect through HPLC, suitably merge elutriant, drying under reduced pressure; namely get ring dipeptides C6; namely six hydrogen-7-hydroxyl-3-(2-methyl-propyl) pyrrolo-[1,2-a) pyrazine-Isosorbide-5-Nitrae-diketone.This compound has reported to have broad-spectrum antibacterial action at present.
Technical scheme of the present invention is as follows:
The Phellinus crude extract is made by following method:
(1) fermentative medium formula in gram/100 milliliter is:
W-Gum 1-5% glucose 1-5%
Peptone 0.1-0.5% yeast extract paste 0.1-0.5%
Sal epsom 0.1-0.5% potassium primary phosphate 0.01-0.05%
(2) the Phellinus bacterial classification is inoculated into ordinary method in the Erlenmeyer flask that liquid nutrient medium is housed, with 20～35 ℃ of temperature, the shaking flask rotating speed is 80～280r/min, and under pH 3～8 conditions, vibrations were cultivated 7～15 days; In cultivation when the pH value drops to 2.5～4, seed in shaking flask is inoculated in the nutrient solution of 50L fermentor tank, with 20～35 ℃ of temperature, fermentor tank pressure 0.1～0.2 kg/cm, pH 3～8, air flow 0.5～1.1vvm, the condition that stirring velocity is 100～280 rev/mins, cultivated 7～15 days, and can utilize the phellinus igniarius mycelium fermentation liquid to prepare the Phellinus crude extract;
(3) get gained phellinus igniarius mycelium fermentation liquid in step (2), with its concentrating under reduced pressure in a usual manner, make its volume be concentrated to 1/2～1/5 of original volume;
(4) be that 60～95% the ethanol fermented liquid after concentrated to above-mentioned steps (3) extracts with volume percent, wherein, the amount that adds ethanol is 2～5 times of concentrated solution volume, can make that in extracting solution, alcohol concn reaches 60～90%;
(5) to step (4) gained extracting solution under 50～70 ℃ of conditions, heated 1～2 hour; Separate with ordinary method, and remove impurity by cascade filtration, separate obtaining ethanol extract; With above-mentioned ethanol extract concentrating under reduced pressure in a usual manner, make its volume be concentrated to 1/5～1/10 of original volume;
(6) just the concentrated solution of step (5) gained carries out drying with the method for frozen drying, gets the Phellinus crude extract.
The method of separating ring dipeptides C6:
The Phellinus bacterium crude extract → purification on normal-phase silica gel chromatography → anti-phase preparation of methyl alcohol gradient elution → HPLC detection → high pressure → HPLC detection → 1D-HNMR → ring dipeptides C6.
Concrete grammar is:
(1) preparation Phellinus bacterium crude extract;
(2) Phellinus bacterium crude extract and the purification on normal-phase silica gel mixing with above-mentioned steps (1) gained stirs, and dry, carries out the silica gel normal phase column chromatography, uses eluent wash-out 2-5 time;
(3) collect last elutriant in above-mentioned steps (2), concentrating under reduced pressure uses dissolve with methanol, and the methanol gel column chromatography is used the eluent wash-out;
(4) elutriant that the step (3) of using HPLC to detect collection obtains suitably merges elutriant, drying under reduced pressure;
(5) product that step (4) is obtained carries out the high pressure reversed phase chromatography, and eluent is methyl alcohol and water;
(6) product that step (5) is obtained carries out HPLC and detects, and drying under reduced pressure is ring dipeptides C6.
The present invention is by the significant advantage of the isolation technique of ring dipeptides C6 in the Phellinus bacterium: present method adopts multiple chromatographic technique to combine, can be exquisite clear and definite to structure, and purity is greater than 95% ring dipeptides C6.Mature technical route is clear and definite, and is accurately efficient.
Description of drawings
Fig. 1 is the structural formula of ring dipeptides C6;
Fig. 2 is the one-dimensional nuclear magnetic resonance H spectrum of ring dipeptides C6.
Embodiment
Example 1:
The Phellinus crude extract is made by following method:
(1) fermentative medium formula in gram/100 milliliter is:
W-Gum 1% glucose 1%
Peptone 0.1% yeast extract paste 0.1%
Sal epsom 0.1% potassium primary phosphate 0.01%
(2) the Phellinus bacterial classification is inoculated into ordinary method in the Erlenmeyer flask that liquid nutrient medium is housed, with 25 ℃ of temperature, the shaking flask rotating speed is 110r/min, and under pH 7 conditions, vibrations were cultivated 7 days; In cultivation when the pH value drops to 3, seed in shaking flask is inoculated in the nutrient solution of 50L fermentor tank, with 25 ℃ of temperature, fermentor tank pressure 0.1 kg/cm, pH 3, air flow 0.5-1.1vvm, the condition that stirring velocity is 100 rev/mins, cultivated 7 days, and can utilize the phellinus igniarius mycelium fermentation liquid to prepare the Phellinus crude extract;
(3) get gained phellinus igniarius mycelium fermentation liquid in step (2), with its concentrating under reduced pressure in a usual manner, make its volume be concentrated to 1/3 of original volume;
(4) be that 70% the ethanol fermented liquid after concentrated to above-mentioned steps (3) extracts with volume percent, wherein, the amount that adds ethanol is 5 times of concentrated solution volume, can make that in extracting solution, alcohol concn reaches 55%;
(5) to step (4) gained extracting solution under 70 ℃ of conditions, heated 1 hour; Separate with ordinary method, and remove impurity by cascade filtration, separate obtaining ethanol extract; With above-mentioned ethanol extract concentrating under reduced pressure in a usual manner, make its volume be concentrated to 1/5 of original volume;
(6) just the concentrated solution of step (5) gained carries out drying with the method for frozen drying, gets the Phellinus crude extract.
Take crude extract 300g and equal-volume 100 order purification on normal-phase silica gel mixings stirrings, carry out the silica gel normal phase column chromatography.The chromatography stationary phase is 200 order purification on normal-phase silica gel, the high 1m of post, and diameter 20cm, eluent is for being respectively chloroform, chloroform: methyl alcohol=75:1 is 4,5 column volumes of wash-out respectively.And with gained elutriant difference called after Fr-1, Fr-2.With the Fr-2 dissolve with methanol, carry out the methanol gel chromatography, eluent is methyl alcohol.Then be that HPCL detects, condition is 0min:0% methyl alcohol, and 10min:100% methyl alcohol, appearance time are 4.95min.Then, carry out the anti-phase preparation of high pressure, A is water mutually, and B is methyl alcohol mutually.Wash-out concentration is the methanol aqueous solution of 25%-75%.collect elutriant, evaporated under reduced pressure, HPLC detects, suitably merge, carry out the 1D-HNMR nuclear magnetic resonance spectroscopy, frequency is 400MHZ, analytical results is δ 7.51 (s, 1H), 4.03 (dd, J=34.9, 28.1 Hz, 2H), 3.81 – 3.33 (m, 2H), 2.36 (d, J=8.8 Hz, 2H), 2.15 – 1.68 (m, 5H), 1.60 (dd, J=10.1, 3.8 Hz, 2H), 0.94 (ddd, J=18.8, 17.2, 9.8 Hz, 7H). prove ring dipeptides C6, i.e. six hydrogen-7-hydroxyl-3-(2-methyl-propyl) pyrrolo-[1, 2-a) pyrazine-1, 4-diketone.
Example 2:
The Phellinus crude extract is made by following method:
(1) fermentative medium formula in gram/100 milliliter is:
W-Gum 3% glucose 2%
Peptone 0.5% yeast extract paste 0.5%
Sal epsom 0.5% potassium primary phosphate 0.05%
(2) the Phellinus bacterial classification is inoculated into ordinary method in the Erlenmeyer flask that liquid nutrient medium is housed, with 30 ℃ of temperature, the shaking flask rotating speed is 180r/min, and under the pH6 condition, vibrations were cultivated 15 days; In cultivation when the pH value drops to 2.5, seed in shaking flask is inoculated in the nutrient solution of 50L fermentor tank, with 30 ℃ of temperature, fermentor tank pressure 0.2 kg/cm, pH 3, air flow 0.5-1.1vvm, the condition that stirring velocity is 180 rev/mins, cultivated 15 days, and can utilize the phellinus igniarius mycelium fermentation liquid to prepare the Phellinus crude extract;
(3) get gained phellinus igniarius mycelium fermentation liquid in step (2), with its concentrating under reduced pressure in a usual manner, make its volume be concentrated to 1/5 of original volume;
(4) be that 90% the ethanol fermented liquid after concentrated to above-mentioned steps (3) extracts with volume percent, wherein, the amount that adds ethanol is 4 times of concentrated solution volume, can make that in extracting solution, alcohol concn reaches 70%;
(5) to step (4) gained extracting solution under 55 ℃ of conditions, heated 2.5 hours; Separate with ordinary method, and remove impurity by cascade filtration, separate obtaining ethanol extract; With above-mentioned ethanol extract concentrating under reduced pressure in a usual manner, make its volume be concentrated to 1/10 of original volume;
(6) just the concentrated solution of step (5) gained carries out drying with the method for frozen drying, gets the Phellinus crude extract.
Take crude extract 100g and equal-volume 100 order purification on normal-phase silica gel mixings stirrings, carry out the silica gel normal phase column chromatography.The chromatography stationary phase is 200-300 order purification on normal-phase silica gel, the high 0.8m of post, and diameter 10cm, eluent is for being respectively chloroform, chloroform: methyl alcohol=75:1 is 4,5 column volumes of wash-out respectively.And with gained elutriant difference called after Fr-1, Fr-2.With the Fr-2 dissolve with methanol, carry out the methanol gel chromatography, eluent is methyl alcohol.Then be that HPCL detects, condition is 0min:0% methyl alcohol, and 10min:100% methyl alcohol, appearance time are 5.10min.Then, carry out the anti-phase preparation of high pressure, A is water mutually, and B is methyl alcohol mutually.Wash-out concentration is the methanol aqueous solution of 25%-70%.collect elutriant, evaporated under reduced pressure, HPLC detects, suitably merge, carry out the 1D-HNMR nuclear magnetic resonance spectroscopy, frequency is 400MHZ, analytical results is δ 7.51 (s, 1H), 4.03 (dd, J=34.9, 28.1 Hz, 2H), 3.81 – 3.33 (m, 2H), 2.36 (d, J=8.8 Hz, 2H), 2.15 – 1.68 (m, 5H), 1.60 (dd, J=10.1, 3.8 Hz, 2H), 0.94 (ddd, J=18.8, 17.2, 9.8 Hz, 7H). prove ring dipeptides C6, i.e. six hydrogen-7-hydroxyl-3-(2-methyl-propyl) pyrrolo-[1, 2-a) pyrazine-1, 4-diketone.
Claims (10)
1. encircle the anti-phase preparation separation method of high pressure of dipeptides C6 in the Phellinus bacterium, its step order is as follows:
(1) preparation Phellinus bacterium crude extract;
(2) Phellinus bacterium crude extract and the purification on normal-phase silica gel mixing with above-mentioned steps (1) gained stirs, and dry, carries out the silica gel normal phase column chromatography, uses eluent wash-out 2-5 time;
(3) collect last elutriant in above-mentioned steps (2), concentrating under reduced pressure uses dissolve with methanol, and the methanol gel column chromatography is used the eluent wash-out;
(4) elutriant that the step (3) of using HPLC to detect collection obtains suitably merges elutriant, drying under reduced pressure;
(5) product that step (4) is obtained carries out the high pressure reversed phase chromatography, and eluent is methyl alcohol and water;
(6) product that step (5) is obtained carries out HPLC and detects, and drying under reduced pressure is ring dipeptides C6.
As claimed in claim 1 a kind of from the Phellinus bacterium method of Separation of Benzene ethanol, it is characterized in that described Phellinus crude extract, made by following method:
(1) fermentative medium formula in gram/100 milliliter is:
W-Gum 1-5% glucose 1-5%
Peptone 0.1-0.5% yeast extract paste 0.1-0.5%
Sal epsom 0.1-0.5% potassium primary phosphate 0.01-0.05%
(2) fermentation culture method of Phellinus crude extract is as claimed in claim 1, the Phellinus bacterial classification is inoculated into ordinary method in the Erlenmeyer flask that liquid nutrient medium is housed, with 20～35 ℃ of temperature, the shaking flask rotating speed is 80～280r/min, under pH 3～8 conditions, vibrations were cultivated 7～15 days; In cultivation when the pH value drops to 2.5～4, seed in shaking flask is inoculated in the nutrient solution of 50L fermentor tank, with 20～35 ℃ of temperature, fermentor tank pressure 0.1～0.2 kg/cm, pH 3～8, air flow 0.5～1.1vvm, the condition that stirring velocity is 100～280 rev/mins, cultivated 7～15 days, and can utilize the phellinus igniarius mycelium fermentation liquid to prepare the Phellinus crude extract;
(3) get gained phellinus igniarius mycelium fermentation liquid in step (2), with its concentrating under reduced pressure in a usual manner, make its volume be concentrated to 1/2～1/5 of original volume;
(4) be that 60～95% the ethanol fermented liquid after concentrated to above-mentioned steps (3) extracts with volume percent, wherein, the amount that adds ethanol is 2～5 times of concentrated solution volume, can make that in extracting solution, alcohol concn reaches 60～90%;
(5) to step (4) gained extracting solution under 50～70 ℃ of conditions, heated 1～2 hour; Separate with ordinary method, and remove impurity by cascade filtration, separate obtaining ethanol extract; With above-mentioned ethanol extract concentrating under reduced pressure in a usual manner, make its volume be concentrated to 1/5～1/10 of original volume;
(6) just the concentrated solution of step (5) gained carries out drying with the method for frozen drying, gets the Phellinus crude extract.
3. the separation method of ring dipeptides C6 in Phellinus bacterium as claimed in claim 1, it is characterized in that described ring dipeptides C6 be six hydrogen-7-hydroxyl-3-(2-methyl-propyl) pyrrolo-[1,2-a) pyrazine-Isosorbide-5-Nitrae-diketone.
4. the separation method of ring dipeptides C6 in Phellinus bacterium as claimed in claim 1, is characterized in that, the sample silica gel of mixing described in step (2) is 100 order purification on normal-phase silica gel, and eluent is chloroform and/or methyl alcohol.
5. the separation method of ring dipeptides C6 in Phellinus bacterium as claimed in claim 1, is characterized in that, the eluent described in step (3) is methyl alcohol.
6. the separation method of ring dipeptides C6 in Phellinus bacterium as claimed in claim 1, is characterized in that, the HPLC condition described in step (4) is 0min:100% water, 10min:100% methyl alcohol.
7. the separation method of ring dipeptides C6 in Phellinus bacterium as claimed in claim 1, is characterized in that, the described reversed material of step (5) is C-18 or C-8.
8. the separation method of ring dipeptides C6 in Phellinus bacterium as claimed in claim 1, is characterized in that, the described eluent of step (5) is methyl alcohol: water=25%-75%.
9. the separation method of ring dipeptides C6 in Phellinus bacterium as claimed in claim 1, is characterized in that, the described high pressure reversed phase chromatography of step (5) number of times is 1-3 time.
10. the separation method of ring dipeptides C6 in Phellinus bacterium as claimed in claim 1, is characterized in that, it is 4.95-5.47min that the described HPLC of step (6) goes out the cutting edge of a knife or a sword time.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004005292A1 (en) * | 2002-07-02 | 2004-01-15 | Universidade De Santiago De Compostela | Novel antibiotics against vibrio anguillarum and the applications thereof in cultures of fish, crustaceans, molluscs and other aquaculture activities |
| WO2007122446A1 (en) * | 2006-04-26 | 2007-11-01 | Commissariat A L'energie Atomique | Cyclodipeptide synthetases and their use for synthesis of cyclo(leu-leu) cyclodipeptide |
| CN102417509A (en) * | 2011-12-28 | 2012-04-18 | 大连美罗中药厂有限公司 | Application of three compounds extracted from starfish to promotion of bone fracture healing, and preparation method for three compounds |
| CN102533600A (en) * | 2011-12-30 | 2012-07-04 | 中国科学院南海海洋研究所 | Marine streptomyces/pyranosesquiterpene compound and preparation method and application thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004005292A1 (en) * | 2002-07-02 | 2004-01-15 | Universidade De Santiago De Compostela | Novel antibiotics against vibrio anguillarum and the applications thereof in cultures of fish, crustaceans, molluscs and other aquaculture activities |
| WO2007122446A1 (en) * | 2006-04-26 | 2007-11-01 | Commissariat A L'energie Atomique | Cyclodipeptide synthetases and their use for synthesis of cyclo(leu-leu) cyclodipeptide |
| CN102417509A (en) * | 2011-12-28 | 2012-04-18 | 大连美罗中药厂有限公司 | Application of three compounds extracted from starfish to promotion of bone fracture healing, and preparation method for three compounds |
| CN102533600A (en) * | 2011-12-30 | 2012-07-04 | 中国科学院南海海洋研究所 | Marine streptomyces/pyranosesquiterpene compound and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| 吴秀丽，等: "火木层孔菌液体培养物的化学成分研究", 《中国中药杂志》 * |
| 李海峰，等: "枯草芽孢杆菌7Ze3环二肽的分离与鉴定", 《江苏农业科学》 * |
| 漆淑华，等: "海洋细菌Ps eudomonas sp.抗菌代谢产物的研究", 《天然产物研究与开发》 * |
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