CN102492605B - Bioreactor and sophorolipid continuous production method by using the same - Google Patents
Bioreactor and sophorolipid continuous production method by using the same Download PDFInfo
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- CN102492605B CN102492605B CN201110388977.1A CN201110388977A CN102492605B CN 102492605 B CN102492605 B CN 102492605B CN 201110388977 A CN201110388977 A CN 201110388977A CN 102492605 B CN102492605 B CN 102492605B
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Abstract
According to a bioreactor and a sophorolipid continuous production method by using the same provided by the invention, as fed-batch of a fermentation broth, a product is recovered at a liquid outlet through intravenous drip. A technology mainly comprises the following steps: centrifuging a fermentation waste liquid by a centrifuge and recovering precipitated yeast byproduct at a lower layer; mixing a centrifuged supernatant and a crude product from intravenous drip and recovering a lower layer product to a secondary purified product gathering tank 2-5 h after settlement layering; disinfecting the secondary purified product by an autoclave and storing directly, or drying at 55-75 DEG C in a drying tower to gain a constant weight and storing.
Description
Technical field
A kind of method that the present invention relates to bio-reactor and produce continuously sophorolipid.
Background technology
When occurrence of large-area oil spill accident, when especially marine weather conditions are severe, use the physical methods such as the oil fence, oil skimmer vessel to reclaim oil spilling very difficult.In this case, oil-spill dispersant can accelerate oily natural evanishment, and under wave effect, it is more easily broken for oil little oil droplet, accelerates the natural biology degraded of oil spilling.The use of oil-spill dispersant is gradually by people are accepted extensively, and most shoreline country all depends on dispersion agent in reply oil leakage.
Oil spill dispersant can be divided into chemical oil spill dispersant and Bio-dispersants.Because the production cost of the main component bio-surfactant of Bio-dispersants is high, hinder the large-scale application of Bio-dispersants, what in the time of reply marine oil overflow accident, use at present is mainly chemical oil spill dispersant.But chemical oil spill dispersant itself has certain toxicity, the therefore application of countries in the world government to chemical oil spill dispersant, all adopts a prudent policy.Bio-dispersants is the substituted chemistry oil spill dispersant trend that is inevitable progressively, and China is also in July, 2010, in the time of the crude oil pollution in reply Dalian, wide-scale adoption first biotechnology process oil spill accident, obtained good effect.
Sophorolipid, as degradable biological tensio-active agent, has the characteristics such as good emulsifying, dispersion, solubilising, and good with human body and Environmental compatibility, and the Bio-dispersants taking it as main component is the favorable substitutes of chemical oil spill dispersant.In addition, sophorolipid has very high research and development and is worth in fields such as food, medicine, makeup, metallurgy, environment protection, waste oil recovery, and oneself is applied to the key areas such as daily use chemicals, medicine, foodstuffs industry, petroleum industry at present.In the world sophorolipid theoretical investigation and application and development are carried out for many years, but China at present in this field also in the starting stage.
The range of application of sophorolipid is boundless, but its production cost is high, limiting sophorolipid and commercially producing on a large scale process, now China is domestic also a large-scale sophorolipid manufacturing enterprise, there is no the public offering of sterling sophorolipid in international coverage yet.The factor of restriction sophorolipid suitability for industrialized production mainly comprises throughput, culture condition, the mode of production and the separation and Extraction mode etc. of bacterial classification, wherein induction mutation of bacterium work and culture condition optimization research are very general, also found comparatively good bacterial classification, under suitable culture condition, can reach higher and stable output, but all in all, existing technique with realize the desired low cost of suitability for industrialized production and also have very large gap.
Summary of the invention
The technical problem to be solved in the present invention is to provide one and has the production of reduction and separation costs, environmental friendliness, the method for continuous production sophorolipid simple to operate.
For solving the problems of the technologies described above, the present invention adopts following technical scheme: comprise tank body, and be arranged on the heating jacket on tank body; In described tank body, be provided with the sieve plate with trapezoidal hole, described sieve plate is the main fermentation region of cylinder above tank body is separated into and is the product deposition region of cone below; The volume ratio of described main fermentation region and product deposition region is between 8:1-12:1, and the aspect ratio in the region of mainly fermenting is between 2:1-3:1; The upper end in described main fermentation region arranges venting port, condensate water circulatory pipeline, inoculation mouth, thief hole, material-feeding port, motor and electrode, is provided with agitator and main breather line in described main fermentation region; Described product is provided with assisted ventilation pipeline in deposition region, and lower end is provided with leakage fluid dram.
Further, described agitator is Dual-layer blade, comprises the froth breaking oar on upper strata and the stirring rake of lower floor.
Further, on described sieve plate, offer isosceles trapezoid hole, sieve plate upper surface percentage of open area be 3%-6%, the percentage of open area of lower surface is 75%, the aperture, upper end in isosceles trapezoid hole is between 2mm-4mm, the aperture, lower end in isosceles trapezoid hole is between 6mm-20mm, and top and bottom, isosceles trapezoid hole aperture ratio is between 1:5-1:3, and plate thickness is 10mm-15mm.
Another technical problem that the present invention will solve is to provide one and has the production of reduction and separation costs, environmental friendliness, bio-reactor simple to operate
For solving the problems of the technologies described above, the present invention adopts following technical scheme: in bio-reactor, add the fermention medium after sterilizing, add cultured seed liquor by inoculation mouth, speed with 50mL/d-80mL/d is added rapeseed oil in batches, the upper lower tank of 55h-96h logical oxygen simultaneously before fermentation, controlling rotating speed remains on more than 50% dissolved oxygen, ferment and ventilate and stop to 96h lower end, after 3h-5h, open valve, discharge sedimentation crude product for quiet, while is with the speed afterfermentation liquid in batches of 40mL/d-100mL/d, 306h-322h finally can continuously ferment, accumulative total reclaims crude product 4.75kg-6.75kg, the production efficiency of unit time unit's fermentor tank volume improves more than 3 times compared with single batch fermentation, effectively fermentation time has extended more than 4 times, after fermented waste fluid is centrifugal, the crude product after getting supernatant liquor and reclaiming mixes, and after mixing, after sedimentation layering 2h, lower floor's product is recovered to secondarily purified product-collecting tank, product after secondary-cleaned, after sterilization tank carries out sterilizing, is dried to constant weight through 70 DEG C, obtains sterling sophorolipid 2.92kg-3.96kg,
Further, ferment for carrying out aerobic fermentation under the condition that is 3.0-4.0 at 25-30 DEG C, pH.
Further, nutrient media components is glucose 130g/L-150g/L, yeast powder 3g/L-10g/L and urea 1g/L.
Further, nutrient media components is glucose 150g/L, yeast powder 3g/L, peptone 0.7g/L, Trisodium Citrate 5g/L, MgSO47H2O4g/L, (NH4) 2SO4 2g/L, KH2PO4 1g/L, NaCl0.1g/L and CaCl22H2O0.1g/L.
Further, described seed liquor is that thalline is cultivated the thalline pregnant solution of 48 hours, culture condition: pH value is 6 through seed culture medium at 30 DEG C; The composition of described seed culture medium is: glucose 100g/L, yeast powder 10g/L and urea 1g/L.
Bio-reactor of the present invention and the method for producing continuously sophorolipid thereof, be intended to, by existing batch fermentation is produced and changed continuous production into, improve the output of sophorolipid; Reduce separation costs, such as the sepn process with respect to traditional, the present invention is not with an organic solvent extraction completely, and environmental friendliness, does not have the loss of product; When lock out operation, can Incidental recovery yeast byproduct, do not need how extra operation and expense; Whole flow operations is simple, and device control is easy.The present invention has realized the continuous production of sophorolipid by designing a kind of new reactor, serve as theme with new separation method simultaneously, design the production technique that efficient, the energy-conservation continuous production of complete sophorolipid, separation and byproduct reclaim, can greatly save production cost, for the large scale continuous prod that realizes sophorolipid provides possibility.
Brief description of the drawings
Fig. 1 is the schematic diagram of bio-reactor of the present invention;
Fig. 2 is the technological process of production block diagram of the present invention's method of producing continuously sophorolipid.
Embodiment
In the present embodiment, with reference to Fig. 1, described bio-reactor, comprises tank body, and is arranged on the heating jacket 14 on tank body; In described tank body, be provided with the sieve plate 17 with trapezoidal hole, described sieve plate 17 is the main fermentation region of cylinder above tank body is separated into and is the product deposition region of cone below; The volume ratio of described main fermentation region and product deposition region is between 8:1-12:1, and the aspect ratio in the region of mainly fermenting is between 2:1-3:1; The upper end in described main fermentation region arranges venting port 10, condensate water circulatory pipeline 11, inoculation mouth 12, thief hole 20, material-feeding port 21, motor 22 and electrode (not shown), is provided with agitator and main breather line 19 in described main fermentation region; Described product is provided with assisted ventilation pipeline 15 in deposition region, and lower end is provided with leakage fluid dram 16.
Wherein, described agitator is Dual-layer blade, comprises the froth breaking oar 13 on upper strata and the stirring rake 18 of lower floor.On described sieve plate 17, offer isosceles trapezoid hole, sieve plate 17 upper surfaces percentage of open area be 3%-6%, the percentage of open area of lower surface is 75%, the aperture, upper end in isosceles trapezoid hole is between 2mm-4mm, the aperture, lower end in isosceles trapezoid hole is between 6mm-20mm, top and bottom, isosceles trapezoid hole aperture ratio is between 1:5-1:3, and plate thickness is 10mm-15mm.Such design be because, upper surface aperture I to be effectively to stop in production process thalline to lower end sedimentation, lower surface aperture is greatly conducive under aeration condition the thalline of deposition and material to be returned the main fermentation region of part; The assisted ventilation pipeline 15 of product deposition region, can, when production in earlier stage provides oxygen for product deposition region, play the stirring action of Accelerative mass transfer, in the fermentation later stage, stop the ventilation of product deposition region, lower end, make product deposition region become for the benefit of relative stagnant zone of product deposition; Meanwhile, the existence of sieve plate 17 can effectively slow down the product vortex in when discharge, makes tank inner fluid in laminar flow regime, when quiet product that reclaims deposition, avoids the unnecessary back-mixing of product and fermented liquid.
The method of as shown in Figure 2, producing continuously sophorolipid comprises following equipment: bio-reactor 1, whizzer 2, thick product-collecting tank 3, yeast holding tank 4, fermented liquid supernatant liquid holding tank 5, secondarily purified product-collecting tank 6, sterilization tank 7, drying plant 8 and sterling hold-up vessel 9.
The method of producing continuously sophorolipid is: taking bio-reactor height overall as 0.4cm, cylinder and conical part aspect ratio are 3:1, right cylinder tank diameter 0.2m, plate thickness 10mm, the upper and lower aperture ratio of trapezoidal hole is 1:3, upper surface aperture is 3mm, useful volume is that the reactor of 10L is specific embodiment, and wherein, main breather line 19 bores are 1cm, assisted ventilation pipeline 15 bores are 0.8cm, all can stop at any time respectively or start up and down ventilation.Production instance is as follows:
Embodiment 1
During taking batch fermentation sophorolipid output as the bacterial strain 1 of 90g/L be example as producing bacterial strain.
To in device, add the fermention medium after 6L sterilizing, nutrient media components is glucose 130g/L, yeast powder 10g/L, urea 1g/L.Adding cultured seed liquor, described seed liquor by inoculation mouth is that thalline is cultivated the thalline pregnant solution of 48 hours, culture condition: pH value is 6 through seed culture medium at 30 DEG C; The composition of described seed culture medium is: glucose 100g/L, yeast powder 10g/L and urea 1g/L; Add rapeseed oil with the speed of 50mL/d more in batches.Speed is added rapeseed oil in batches.Before fermentation, the upper lower tank of 96h logical oxygen simultaneously, controls rotating speed dissolved oxygen is remained on more than 50%.Ferment and ventilate and stop to 96h lower end, open valve after 5h, discharge sedimentation crude product for quiet, the while is with the speed afterfermentation liquid in batches of 40mL/d.The 306h that finally can continuously ferment, accumulative total reclaims crude product 4.75kg.The production efficiency of unit time unit's fermentor tank volume improves more than 3 times compared with single batch fermentation, and effectively fermentation time has extended more than 4 times.Wherein, ferment for carrying out aerobic fermentation under the condition that is 3.0-4.0 at 25-30 DEG C, pH.
After fermented waste fluid is centrifugal, the crude product after getting supernatant liquor and reclaiming mixes, and after mixing, after sedimentation layering 2h, lower floor's product is recovered to secondarily purified product-collecting tank; Product after secondary-cleaned, after sterilization tank carries out sterilizing, is dried to constant weight through 70 DEG C, obtains sterling sophorolipid 2.92kg.Adopt the more traditional organic solvent extraction separating technology of this separating technology to save the ethyl acetate of 6 liters, and greatly reduce relevant distillation energy consumption and condensation water, separation costs is only 1/4 left and right of traditional technology.Meanwhile, obtain 0.26kg yeast byproduct.
Embodiment 2
During taking batch fermentation sophorolipid output as the bacterial strain 2 of 120g/L be example as producing bacterial strain.
To in device, add the fermention medium after 6L sterilizing, nutrient media components is glucose 150g/L, yeast powder 3g/L, peptone 0.7g/L, Trisodium Citrate 5g/L, MgSO47H2O4g/L, (NH4) 2SO42g/L, KH2PO4 1g/L, NaCl0.1g/L, CaCl22H2O0.1g/L.Adding cultured seed liquor, described seed liquor by inoculation mouth is that thalline is cultivated the thalline pregnant solution of 48 hours, culture condition: pH value is 6 through seed culture medium at 30 DEG C; The composition of described seed culture medium is: glucose 100g/L, yeast powder 10g/L and urea 1g/L; Add rapeseed oil with the speed of 80mL/d more in batches.Before fermentation, the upper lower tank of 55h logical oxygen simultaneously, controls rotating speed and makes dissolved oxygen remain on higher level.Ferment and ventilate and stop to 96h lower end, open valve after 3h, discharge sedimentation crude product for quiet, the while is with the speed afterfermentation liquid in batches of 100mL/d.The 322h that finally can continuously ferment, accumulative total reclaims crude product 6.75kg.The production efficiency of unit time unit's fermentor tank volume improves 3.5 times compared with single batch fermentation, and effectively fermentation time has extended more than 4 times.
After fermented waste fluid is centrifugal, the crude product after getting supernatant liquor and reclaiming mixes, and after mixing, after sedimentation layering 2h, lower floor's product is recovered to secondarily purified product-collecting tank; Product after secondary-cleaned, after sterilization tank carries out sterilizing, is dried to constant weight through 70 DEG C, obtains sterling sophorolipid 3.96kg.Adopt the more traditional organic solvent extraction separating technology of this separating technology to save the ethyl acetate of 8 liters, and greatly reduce relevant distillation energy consumption and condensation water, separation costs is only 1/4 left and right of traditional technology.Meanwhile, obtain 0.35kg yeast byproduct.
Bio-reactor of the present invention and the method for producing continuously sophorolipid thereof, advantage is: by designing a kind of novel bio-reactor, the mode of production of existing batch fermentation is changed into the continuous mode of production, greatly improved the utilization ratio of substratum, the service efficiency of equipment; By utilizing fermented supernatant fluid again to mix with crude product, be further purified sophorolipid, because fermented supernatant fluid is the saturated solution of sophorolipid, so reduced the loss of product, simultaneously than the traditional method with an organic solvent extracting, due to not with an organic solvent extraction completely, energy efficient of the present invention, environmental friendliness, with low cost; When lock out operation, can Incidental recovery yeast byproduct, do not need how extra operation and expense; The whole technological process of production is complete, and operation steps is simple, and device control is easy.
Below the present invention is described in detail, the above, be only the present invention's preferred embodiment, when not limiting the scope of the present invention, allly does impartial variation and modification according to the application's scope, all should still belong in covering scope of the present invention.
Claims (1)
1. a bio-reactor, is characterized in that: comprise tank body, and be arranged on the heating jacket on tank body; In described tank body, be provided with the sieve plate with trapezoidal hole, described sieve plate is the main fermentation region of cylinder above tank body is separated into and is the product deposition region of cone below; The volume ratio of described main fermentation region and product deposition region is 8: 1-12: between 1, the aspect ratio in the region of mainly fermenting is 2: 1-3: between 1; The upper end in described main fermentation region arranges venting port, condensate water circulatory pipeline, inoculation mouth, thief hole, material-feeding port, motor and electrode, is provided with agitator and main breather line in described main fermentation region; Described product is provided with assisted ventilation pipeline in deposition region, lower end is provided with leakage fluid dram, described agitator is Dual-layer blade, comprise the froth breaking oar on upper strata and the stirring rake of lower floor, on described sieve plate, offer isosceles trapezoid hole, the percentage of open area of sieve plate upper surface is 3%-6%, the percentage of open area of lower surface is 75%, the aperture, upper end in isosceles trapezoid hole is between 2mm-4mm, the aperture, lower end in isosceles trapezoid hole is between 6mm-20mm, top and bottom, isosceles trapezoid hole aperture ratio is 1: 5-1: between 3, plate thickness is 10mm-15mm.
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| JP6697662B2 (en) * | 2015-11-09 | 2020-05-27 | アライドカーボンソリューションズ株式会社 | Oil recovery method using sophorolipid |
| CN105999763B (en) * | 2016-05-20 | 2018-01-23 | 天津大学 | A kind of sophorolipid fermenting organism piece-rate system and method |
| GB201610932D0 (en) * | 2016-06-22 | 2016-08-03 | Univ Of Manchester The | Improvements in and relating to lipid production |
| CN107254388A (en) * | 2017-08-14 | 2017-10-17 | 合肥明英富海生物科技有限公司 | Installation for fermenting |
| CN108546631A (en) * | 2018-04-11 | 2018-09-18 | 重庆工业职业技术学院 | A kind of decomposition apparatus of biomaterial |
| EP3814478A4 (en) * | 2018-06-29 | 2022-04-13 | Locus IP Company, LLC | Method and apparatus for continuous production of sophorolipids |
| CN114350533A (en) * | 2022-01-05 | 2022-04-15 | 华东理工大学 | Adaptive evolution method for improving resistance of saccharomyces cerevisiae to 2-pe and chemostatic culture device |
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| US5900366A (en) * | 1992-06-18 | 1999-05-04 | Institut Français Du Petrole | Production of sophorolipid acetate acids from oils or esters |
| CN2865844Y (en) * | 2005-08-15 | 2007-02-07 | 中国石油化工股份有限公司 | Distributor of axial flow adiabatic fixed-bed reactor |
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| US5900366A (en) * | 1992-06-18 | 1999-05-04 | Institut Français Du Petrole | Production of sophorolipid acetate acids from oils or esters |
| CN2865844Y (en) * | 2005-08-15 | 2007-02-07 | 中国石油化工股份有限公司 | Distributor of axial flow adiabatic fixed-bed reactor |
| CN101066805A (en) * | 2007-06-05 | 2007-11-07 | 浙江大学 | Efficient anaerobic ammoxidation reactor |
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| CN202322821U (en) * | 2011-11-30 | 2012-07-11 | 中海石油环保服务（天津）有限公司 | Biological reactor |
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