CN205382166U - Use fermentation and foam separation coupling production polymyxins E's device - Google Patents
Use fermentation and foam separation coupling production polymyxins E's device Download PDFInfo
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- CN205382166U CN205382166U CN201620169124.7U CN201620169124U CN205382166U CN 205382166 U CN205382166 U CN 205382166U CN 201620169124 U CN201620169124 U CN 201620169124U CN 205382166 U CN205382166 U CN 205382166U
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- JORAUNFTUVJTNG-BSTBCYLQSA-N n-[(2s)-4-amino-1-[[(2s,3r)-1-[[(2s)-4-amino-1-oxo-1-[[(3s,6s,9s,12s,15r,18s,21s)-6,9,18-tris(2-aminoethyl)-3-[(1r)-1-hydroxyethyl]-12,15-bis(2-methylpropyl)-2,5,8,11,14,17,20-heptaoxo-1,4,7,10,13,16,19-heptazacyclotricos-21-yl]amino]butan-2-yl]amino]-3-h Polymers CC(C)CCCCC(=O)N[C@@H](CCN)C(=O)N[C@H]([C@@H](C)O)CN[C@@H](CCN)C(=O)N[C@H]1CCNC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCN)NC(=O)[C@H](CCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CCN)NC1=O.CCC(C)CCCCC(=O)N[C@@H](CCN)C(=O)N[C@H]([C@@H](C)O)CN[C@@H](CCN)C(=O)N[C@H]1CCNC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCN)NC(=O)[C@H](CCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CCN)NC1=O JORAUNFTUVJTNG-BSTBCYLQSA-N 0.000 claims description 23
- YKQOSKADJPQZHB-YNWHQGOSSA-N n-[(2s)-4-amino-1-[[(2s,3r)-1-[[(2s)-4-amino-1-oxo-1-[[(3s,6s,9s,12s,15r,18s,21s)-6,9,18-tris(2-aminoethyl)-3-[(1s)-1-hydroxyethyl]-12,15-bis(2-methylpropyl)-2,5,8,11,14,17,20-heptaoxo-1,4,7,10,13,16,19-heptazacyclotricos-21-yl]amino]butan-2-yl]amino]-3-h Polymers CCC(C)CCCC(=O)N[C@@H](CCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCN)C(=O)N[C@H]1CCNC(=O)[C@H]([C@H](C)O)NC(=O)[C@H](CCN)NC(=O)[C@H](CCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CCN)NC1=O YKQOSKADJPQZHB-YNWHQGOSSA-N 0.000 claims description 23
- XDJYMJULXQKGMM-UHFFFAOYSA-N polymyxin E1 Natural products CCC(C)CCCCC(=O)NC(CCN)C(=O)NC(C(C)O)C(=O)NC(CCN)C(=O)NC1CCNC(=O)C(C(C)O)NC(=O)C(CCN)NC(=O)C(CCN)NC(=O)C(CC(C)C)NC(=O)C(CC(C)C)NC(=O)C(CCN)NC1=O XDJYMJULXQKGMM-UHFFFAOYSA-N 0.000 claims description 23
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The utility model relates to a microbial fermentation and foam separation technique coupling production polymyxins E's combination unit, the device includes bioreactor, membrane filter, foam separation tower, bleeder valve, by pass vavle, peristaltic pump, bioreactor include: fermentation cylinder, constant temperature test the speed magnetic stirrers, head tank, pH meter, air -blowing pump, feed valve, vacuum pump, wherein, membrane filter sets up in the right side below of fermentation cylinder jar wall discharge gate department, and the material passes through the peristaltic pump through the discharge gate through the bleeder valve and links to each other with foam separation tower right side feed inlet after membrane filter's filtration in the fermentation cylinder, the bottom of foam separation tower links to each other from the feed inlet of backward flow antilinear by pass vavle with the fermentation cylinder. The utility model discloses the device is simple, and foam separation operating conditions is gentle, required cost reduction, waste liquid toxicological harmless, pollution -free, green that the fermentation finishes.
Description
Technical field
This utility model relates to technical field of bioengineering, couples the device producing polymyxin E with foam separation particularly to a kind of application of fermentation.
Background technology
The characteristic that polymyxin E has better antimicrobial spectrum, efficient, low toxicity, residual are few, can be used for feed additive, promote birds and beasts growth, improve efficiency of feed utilization, and be possible to prevent that feedstuff often occurs in large-scale production by colon bacillus and the salmonella-polluted disease caused, polymyxin E has been classified as national legal kind by China, but product titer is relatively low, relies primarily on import.This product is mainly prepared by fermentation method, and traditional separation method mainly uses ion exchange chromatography, and the energy consumption of this process is very big.This seminar adopts the polymyxin E in the method separation and fermentation liquid of foam separation to deliver patent, first fermentation liquor pretreatment, vacuum drying, spray drying obtains the concentrated solution of polymyxin E then, cycles of concentration reaches 15, and yield is 95%, has wide and practical economic effect.
But this method is during the fermentation along with the increase of product can cause serious inhibitory action so that in fermentation liquid, the total titer of polymyxin E is relatively low.The device of foam separation is adopted for this phenomenon existing document at present, it is equipped with bubble distributer at fermenter base, by Bubble fractionation purpose product, this method can effectively eliminate product inhibition, improve productivity, have greatly improved than traditional method, but in foam separation process, along with the rising of foam phase can take away part nutrient substance and cell, reduce raw material availability, reduce concentration ratio, simultaneously because be easily separated a large amount of bubbles of generation in the fermentation tank serve as reactor can affect mass transfer.Accordingly, it would be desirable to design a kind of new device to overcome Problems existing.
Utility model content
Current techniques is coupled with in tank a lot of limitation, the optimum condition of fermentation should be met for operating condition on the one hand and meet the requirement of separation again；On the other hand along with the generation of bubble has taken away product incessantly and can be taken out of by nutrient substance.Based on above limitation, this utility model provides a kind of application of fermentation to couple the device producing polymyxin E with foam separation.This device utilizes peristaltic pump that by film filter, fermentation liquid is entered foam separating tower, product and liquid, and by film, cell is trapped within fermentation tank relaying supervention ferment, it is ensured that thalline constant；At foam separation tower bottom equipped with bubble distributer, produce bubble by air-blowing and carry out foam separation, owing to entering foam separating tower by film filter part nutrient substance, so what this utility model adopted is that the remaining liquid after foam separation is refluxed back in fermentation tank by the mode of circulating reflux, it is ensured that making full use of of nutrient substance；This utility model adopts coupling outside tank body simultaneously, and the operating condition of fermentation and the operating condition of foam separation control respectively, can meet the operating condition of the best.
The technical solution adopted in the utility model is:
A kind of fermentable couples the combined unit producing polymyxin E with foam separating technology, including bioreactor, film filter, foam separating tower, outlet valve, reflux inlet, peristaltic pump；
Described bioreactor includes: fermentation tank, constant temperature test the speed magnetic stirring apparatus, head tank, pH meter, air-blowing pump, inlet valve, vacuum pump；
Described film filter is arranged on the discharge outlet in fermentation tank, is specially an inorfil element film, and 0.45 μm, Tianjin Dong Ke instrument and equipment company limited buys；
Wherein, film filter is arranged on the lower right-hand side tank skin discharge outlet of fermentation tank, and in fermentation tank, material is after the filtration of film filter, is connected with charging aperture on the right side of foam separating tower by peristaltic pump through outlet valve by discharging opening；The bottom of foam separating tower is connected with the charging aperture of fermentation tank from refluxing opening through reflux inlet；
Wherein head tank is connected with lower section on the left of fermentation tank by inlet valve；Described fermentation tank discharging opening is connected with foam separating tower charging aperture；Fermentation tank charging aperture is connected with foam separating tower refluxing opening；
Described foam separating tower includes: foam separation king-post, bubble distributer, froth breaking liquid, water bath with thermostatic control water inlet, water bath with thermostatic control outlet；
Foam separating tower column bottom is connected to bubble distributer and refluxing opening；Described refluxing opening is connected with fermentation tank charging aperture through reflux inlet；Described charging aperture is connected with fermentation tank discharging opening through peristaltic pump；Described foam separation top of tower is provided with product and receives device froth breaking liquid；Described water bath with thermostatic control water inlet (is arranged on lower section on the left of foam separation king-post outer side sleeve, water bath with thermostatic control outlet is arranged on top on the right side of foam separation king-post outer side sleeve, and water bath with thermostatic control water inlet is connected with digital display water bath with thermostatic control with water bath with thermostatic control outlet.
Existing fermentation integrated separation technology, ion exchange resin extracts needs regularly to change resin, relatively costly；And substantial amounts of nutrient substance can be taken out of when foam separation, cause that utilization rate of raw materials and concentration ratio are relatively low.
Relative to existing fermentation process coupled with separation technology, the beneficial effects of the utility model are:
1. reduce cost, environmental protection.Adopting the fermentation in this utility model to carry out the fermentation of polymyxin E, product separation process with foam separation coupling device, device is simple, and foam separation operating condition is gentle, and required cost reduces, the waste liquid nonhazardous of fermentation ends, pollution-free, environmental protection.
2. reduce Product inhibiton, improve utilization rate of raw materials.This utility model removes product polymyxin E online, reduces or relieves the serious product inhibition caused along with product increase during the fermentation, improve the total titer of polymyxin E, compare titer than traditional zymotic and improve about 22%；The nutrient substance being entered foam separating tower by film filter is passed back into fermentation tank by the mode of application circulating reflux, reduces raw-material loss, improves utilization rate of raw materials.
Accompanying drawing explanation
Fig. 1 is that bacillus polymyxa fermentation couples the process flow diagram producing polymyxin E with foam separation.
Wherein, 1-bioreactor；2-film filter；3-foam separating tower；4-outlet valve；5-reflux inlet；6-peristaltic pump.
Fig. 2 is the structural representation of bioreactor
Wherein, 100 fermentation tank；101 constant temperature test the speed magnetic stirring apparatus；102 head tanks；103 discharging openings；104 charging apertures；105 pH meters；106 air-blowing pumps；107 inlet valves；108 vacuum pumps.
Fig. 3 is the structural representation of foam separating tower
Wherein, 301 foam separation king-post；302 charging apertures；303 refluxing openings；304 bubble distributers；305 froth breaking liquid；306 water bath with thermostatic control water inlets；307 water bath with thermostatic control outlets.
Detailed description of the invention
Detailed description of the invention of the present utility model describes in detail as follows with reference to attached Figure 13, but is only used as explanation, is not restriction this utility model.
A kind of fermentable couples the combined unit producing polymyxin E, bioreactor and the foam separator circulation reflux device connected and composed by peristaltic pump with foam separating technology；
The composition of this device such as Fig. 1, including bioreactor (1), film filter (2), foam separating tower (3), outlet valve (4), reflux inlet (5), peristaltic pump (6)；
Composition such as Fig. 2 of described bioreactor (1), including: fermentation tank (100), constant temperature test the speed magnetic stirring apparatus (101), head tank (102), discharging opening (103), charging aperture (104), pH meter (105), air-blowing pump (106), inlet valve (107), vacuum pump (108)；
Described film filter (2) is arranged on discharging opening (103) place in fermentation tank (100), is specially an inorfil element film, and 0.45 μm, Tianjin Dong Ke instrument and equipment company limited buys；
Wherein, film filter (2) is arranged on lower right-hand side tank skin discharging opening (103) place of fermentation tank (100), in fermentation tank (100), material is after the filtration of film filter (2), is connected with foam separating tower (3) right side charging aperture by peristaltic pump (6) through outlet valve (4) by this outlet；The bottom of foam separating tower (3) is connected with the charging aperture (104) above the right side of fermentation tank (100) from refluxing opening through reflux inlet (5)；
Wherein head tank (102) is connected with lower section, fermentation tank (100) left side by inlet valve (107)；Described fermentation tank (100) discharging opening (103) is connected with foam separating tower charging aperture (302)；Fermentation tank (100) charging aperture (104) is connected with foam separating tower refluxing opening (303)；
Described foam separating tower (3) includes again: foam separation king-post (301), charging aperture (302), refluxing opening (303), bubble distributer (304), froth breaking liquid (305), water bath with thermostatic control water inlet (306), water bath with thermostatic control outlet (307)；
The internal diameter of wherein said foam separating tower (301) is 5cm, be highly 75cm for cylinder；Foam separation king-post (301) bottom is connected to bubble distributer (304) and refluxing opening (303)；Described bubble distributer (304) is connected with air-blowing pump；Described refluxing opening (303) is connected with fermentation tank (100) charging aperture (104) through reflux inlet (5)；Described charging aperture (302) is connected with fermentation tank (100) discharging opening (103) through peristaltic pump (6)；Described foam separation top of tower is provided with product and receives device froth breaking liquid (305)；Described water bath with thermostatic control water inlet (306) is arranged on lower section on the left of foam separation king-post outer side sleeve, water bath with thermostatic control outlet (307) is arranged on top on the right side of foam separation king-post outer side sleeve, and water bath with thermostatic control water inlet (306) is connected with digital display water bath with thermostatic control with water bath with thermostatic control outlet (307).
Described outlet valve (4) left side is connected with the film filter discharging opening (103) in fermentation tank (100), and right side is connected with foam separating tower inlet valve (302) by peristaltic pump；
Described reflux inlet (5) left side connects fermentation tank 100 charging aperture (104), and right side is connected with refluxing opening at the bottom of foam separating tower (303).
When fermentation and foam separation coupling operation, first turn on vacuum pump (108), make whole system be in high vacuum state.It is then shut off vacuum pump (108), opening inlet valve (107) makes feed liquid automatically be passed in the fermentation tank (100) that high temperature sterilize processes by head tank (102) under external atmosphere pressure, seed liquor is inoculated in fermentation tank (100) with the inoculum concentration of 10%.Regulate so that fermentation tank (100) liquid amount is 60%-85%, cultivation temperature 30-35 DEG C, speed of agitator 100-400r/min, ventilation 100-300mL/min.With this understanding after fermentation 30-40h, open the digital display constant temperature water bath apparatus outside outlet valve (4), peristaltic pump (6) and knockout tower and water bath with thermostatic control water inlet (306) and water bath with thermostatic control outlet (307), entering in foam separating tower by film filter with the charging rate of 100-300mL/h by fermentation liquid, biological cell is trapped within fermentation tank relaying supervention ferment.When, after certain time, liquid reaches about the 25%-40% that tower body is long-pending in knockout tower, opening foam separation system, be 50-150mL/min at ventilation, temperature is 30-35 DEG C and carries out foam separation, and it is the concentrated solution of polymyxin E that knockout tower top receives liquid.Then the liquid amount for keeping knockout tower is constant, opens reflux inlet (5) and flows back in fermentation tank (100) by liquid more than the foam separation containing nutrient substance, and circulation carries out, until fermentation ends.
Embodiment 1
The bacillus polymyxa bacterium solution obtained from freezing glycerol pipe is inoculated in slant medium and activates, 30 DEG C, cultivate 72h.Bacterium solution after activation is inoculated in seed culture medium, cultivates 24h at 32 DEG C, obtain seed liquor (following example are same).Being inoculated into seed liquor in the fermentation tank (100) of the 1.8L of high temperature sterilize process with the inoculum concentration of 10%, fermentation tank (100) liquid amount is 1.2L, cultivation temperature 30 DEG C, speed of agitator 200r/min, ventilation 150mL/min.Fermentation medium consists of: glucose 20g/L, peptone 10g/L, sodium chloride 1g/L, potassium dihydrogen phosphate 0.1g/L, ammonium sulfate 10g/L, ferrous sulfate 0.1g/L, and it is 7,115-121 DEG C of high pressure steam sterilization 15-30min that ammonia regulates initial pH.With this understanding after fermentation 33h, detection thalline content reaches 4.19g/L, and polymyxin E content reaches 4680U/mL.Now open the digital display constant temperature water bath apparatus outside outlet valve (4), charging peristaltic pump (6) and knockout tower and water bath with thermostatic control water inlet (306) and water bath with thermostatic control outlet (307), entering in foam separating tower by film filter with the charging rate of 200mL/h by fermentation liquid, biological cell is trapped within fermentation tank (100) relaying supervention ferment.When, after 1.5h, liquid reaches 300mL in knockout tower, opening foam separation system, at 32 DEG C, ventilation 50mL/min carries out foam separation, and knockout tower top receives the concentrated solution that liquid is polymyxin E and is froth breaking liquid.Then the liquid amount for keeping knockout tower is constant, opens reflux inlet (5) and flows back in fermentation tank (100) by liquid more than the foam separation containing nutrient substance, and circulation carries out 14h, until fermentation ends.Fermentation ends records total titer and improves 21% than before coupling, and concentrated solution concentration ratio is 3.5, and detecting glucose content in froth breaking liquid by DNS method is 1.322g/L, it was shown that utilization rate of raw materials is high；Fermentation and separation process are not added with virulent material, and itself does not produce new polluter；Applying this appliance arrangement simple, small investment, energy consumption is low can reduce separation costs.
Embodiment 2
Being inoculated into seed liquor in the fermentation tank (100) of the 1.8L of high temperature sterilize process with the inoculum concentration of 10%, fermentation tank (100) liquid amount is 1.4L, cultivation temperature 34 DEG C, speed of agitator 300r/min, ventilation 200mL/min.Fermentation medium consists of: glucose 20g/L, peptone 10g/L, sodium chloride 1g/L, potassium dihydrogen phosphate 0.1g/L, ammonium sulfate 10g/L, ferrous sulfate 0.1g/L, and it is 7,115-121 DEG C of high pressure steam sterilization 15-30min that ammonia regulates initial pH.With this understanding after fermentation 36h, detection thalline content reaches 4.01g/L, and polymyxin E content reaches 4712U/mL.Now open outside outlet valve (4), charging peristaltic pump (6) and knockout tower digital display constant temperature water bath apparatus and water bath with thermostatic control water inlet (306) and water bath with thermostatic control outlet (307), entering in foam separating tower by film filter with the charging rate of 200mL/h by fermentation liquid, biological cell is trapped within fermentation tank (100) relaying supervention ferment.When, after 1.5h, liquid reaches 300mL in knockout tower, opening foam separation system, at 32 DEG C, ventilation 100mL/min carries out foam separation, and it is the concentrated solution of polymyxin E that knockout tower top receives liquid.Then the liquid amount for keeping knockout tower is constant, opens reflux inlet (5) and flows back in fermentation tank (100) by liquid more than the foam separation containing nutrient substance, and circulation carries out 12h, until fermentation ends.Fermentation ends records total titer and improves 23% than before coupling, and concentrated solution concentration ratio is 3.2.
Embodiment 3
Being inoculated into seed liquor in the fermentation tank (100) of the 1.8L of high temperature sterilize process with the inoculum concentration of 10%, fermentation tank (100) liquid amount is 1.3L, cultivation temperature 32 DEG C, speed of agitator 250r/min, ventilation 200mL/min.Fermentation medium consists of: glucose 20g/L, peptone 10g/L, sodium chloride 1g/L, potassium dihydrogen phosphate 0.1g/L, ammonium sulfate 10g/L, ferrous sulfate 0.1g/L, and it is 7,115-121 DEG C of high pressure steam sterilization 15-30min that ammonia regulates initial pH.With this understanding after fermentation 33h, detection thalline content reaches 4.24g/L, and polymyxin E content reaches 4901U/mL.Now open outside outlet valve (4), charging peristaltic pump (6) and knockout tower digital display constant temperature water bath apparatus and water bath with thermostatic control water inlet (306) and water bath with thermostatic control outlet (307), entering in foam separating tower by film filter with the charging rate of 200mL/h by fermentation liquid, biological cell is trapped within fermentation tank (100) relaying supervention ferment.When, after 1.5h, liquid reaches 300mL in knockout tower, opening foam separation system, at 32 DEG C, ventilation 100mL/min carries out foam separation, and it is the concentrated solution of polymyxin E that knockout tower top receives liquid.Then the liquid amount for keeping knockout tower is constant, opens reflux inlet (5) and flows back in fermentation tank (100) by liquid more than the foam separation containing nutrient substance, and circulation carries out 14h, until fermentation ends.Fermentation ends records total titer and improves 22% than before coupling, and concentrated solution concentration ratio is 3.3.
This utility model unaccomplished matter is known technology.
Claims (2)
1. fermentable couples the combined unit producing polymyxin E with foam separating technology, it is characterized by include bioreactor, film filter, foam separating tower, outlet valve, reflux inlet, peristaltic pump；
Described bioreactor includes: fermentation tank, constant temperature test the speed magnetic stirring apparatus, head tank, pH meter, air-blowing pump, inlet valve, vacuum pump；
Wherein, film filter is arranged on the lower right-hand side tank skin discharge outlet of fermentation tank, and in fermentation tank, material is after the filtration of film filter, is connected with charging aperture on the right side of foam separating tower by peristaltic pump through outlet valve by discharging opening；The bottom of foam separating tower is connected with the charging aperture of fermentation tank from refluxing opening through reflux inlet；
Wherein head tank is connected with lower section on the left of fermentation tank by inlet valve；Described fermentation tank discharging opening is connected with foam separating tower charging aperture；Fermentation tank charging aperture is connected with foam separating tower refluxing opening；
Described foam separating tower includes: foam separation king-post, bubble distributer, froth breaking liquid, water bath with thermostatic control water inlet, water bath with thermostatic control outlet；
Foam separating tower column bottom is connected to bubble distributer and refluxing opening；Described refluxing opening is connected with fermentation tank charging aperture through reflux inlet；Described charging aperture is connected with fermentation tank discharging opening through peristaltic pump；Described foam separation top of tower is provided with product and receives device froth breaking liquid；Described water bath with thermostatic control water inlet is arranged on lower section on the left of foam separation king-post outer side sleeve, and water bath with thermostatic control outlet is arranged on top on the right side of foam separation king-post outer side sleeve, and water bath with thermostatic control water inlet is connected with digital display water bath with thermostatic control with water bath with thermostatic control outlet.
2. fermentable as claimed in claim 1 couples the combined unit producing polymyxin E with foam separating technology, it is characterized by the cellulose inoranic membrane that microporous filter membrane material is 0.45 μm used in described film filter.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108559767A (en) * | 2018-01-25 | 2018-09-21 | 江苏凌云药业股份有限公司 | A kind of method that mixed pantoic acid lactone is continuously split and its device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108559767A (en) * | 2018-01-25 | 2018-09-21 | 江苏凌云药业股份有限公司 | A kind of method that mixed pantoic acid lactone is continuously split and its device |
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