CN107964506A - A kind of feeding medium during fermentation Optimal Control System and method - Google Patents
A kind of feeding medium during fermentation Optimal Control System and method Download PDFInfo
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- 238000000855 fermentation Methods 0.000 title claims abstract description 106
- 230000004151 fermentation Effects 0.000 title claims abstract description 106
- 238000000034 method Methods 0.000 title claims abstract description 35
- 239000006052 feed supplement Substances 0.000 claims abstract description 71
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 28
- 239000008103 glucose Substances 0.000 claims abstract description 28
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims abstract description 23
- 230000003287 optical effect Effects 0.000 claims abstract description 15
- 238000012544 monitoring process Methods 0.000 claims abstract description 10
- 241000894006 Bacteria Species 0.000 claims description 13
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 13
- 239000001301 oxygen Substances 0.000 claims description 13
- 229910052760 oxygen Inorganic materials 0.000 claims description 13
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 8
- 241001052560 Thallis Species 0.000 claims description 6
- 239000007789 gas Substances 0.000 claims description 5
- 230000006698 induction Effects 0.000 claims description 5
- 230000000241 respiratory effect Effects 0.000 claims description 5
- 238000003556 assay Methods 0.000 claims description 4
- 239000001569 carbon dioxide Substances 0.000 claims description 4
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 4
- 239000013589 supplement Substances 0.000 claims description 4
- 235000006286 nutrient intake Nutrition 0.000 claims description 3
- 235000016709 nutrition Nutrition 0.000 claims description 3
- 239000002028 Biomass Substances 0.000 claims description 2
- 230000005611 electricity Effects 0.000 claims description 2
- 235000009754 Vitis X bourquina Nutrition 0.000 claims 1
- 235000012333 Vitis X labruscana Nutrition 0.000 claims 1
- 240000006365 Vitis vinifera Species 0.000 claims 1
- 235000014787 Vitis vinifera Nutrition 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 abstract description 6
- 102000004169 proteins and genes Human genes 0.000 abstract description 6
- 239000002609 medium Substances 0.000 description 34
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical class [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 12
- 229940041514 candida albicans extract Drugs 0.000 description 8
- 230000002572 peristaltic effect Effects 0.000 description 8
- 239000012138 yeast extract Substances 0.000 description 8
- 239000013530 defoamer Substances 0.000 description 6
- 235000019341 magnesium sulphate Nutrition 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- 235000013619 trace mineral Nutrition 0.000 description 6
- 239000011573 trace mineral Substances 0.000 description 6
- 239000012137 tryptone Substances 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 5
- 238000005070 sampling Methods 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 239000006143 cell culture medium Substances 0.000 description 4
- 108010046845 tryptones Proteins 0.000 description 4
- 239000000463 material Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 235000005979 Citrus limon Nutrition 0.000 description 2
- 244000131522 Citrus pyriformis Species 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical class [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 235000001727 glucose Nutrition 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019799 monosodium phosphate Nutrition 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000011218 seed culture Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical class [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000012366 Fed-batch cultivation Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000013178 mathematical model Methods 0.000 description 1
- 238000011169 microbiological contamination Methods 0.000 description 1
- 235000021049 nutrient content Nutrition 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/48—Automatic or computerized control
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- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
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Abstract
The invention discloses a kind of feeding medium during fermentation Optimal Control System and method, the system comprises fermentation tank, feed supplement bottle, for monitoring in the fermentation tank glucose electrode of sugared concentration and optical density electrode for monitoring thalline OD in the fermentation tank in real time in real time, and the external feed supplement for being pumped into supplemented medium pumps and external computerized control system, it connects the glucose electrode, optical density electrode and feed supplement pump, according to the sugared concentration, thalline OD and the specific growth rate of setting control the speed of service of the feed supplement pump, supplemented medium is supplemented from the feed supplement bottle to the fermentation tank in the form of the fed-batch mode and continuous stream for controlling specific growth rate add.The present invention system and method, it is possible to increase feeding medium during fermentation automation process and be conducive to improve thalline fermentation density, improve destination protein expression.
Description
Technical field
The present invention relates to fermentation technical field, more particularly to a kind of feeding medium during fermentation Optimal Control System and method.
Background technology
Fed-batch fermentation (Fed-batch fermentation), also referred to as semicontinuous fermentation, refer in batch fermentation mistake
The method that fresh culture or some nutriments are intermittently or continuously added in journey.Application range is very extensive at present, almost
Throughout whole fermentation industry.The theoretical research of fed-batch fermentation almost a blank before the 1970s, in early stage
Industrial production in, the mode of feed supplement is very simple, and intermittent flow adduction constant speed stream is just limited only to by the way of and is added, control
Fermentation is also based on experience.Until Japanese scholars Yoshida in 1973 et al. proposes " fed-batch fermentation " this term,
And first mathematical model is theoretically established, the research of fed-batch fermentation just enters theoretical research stage.Thereafter, with
Research deepens continuously, and fed-batch fermentation achieves great progress at three aspects.In recent years more with theoretical research and
Application deepens continuously, and the content of fed-batch cultivation greatly enriches.
Fed-batch fermentation is by controlling residual sugar content or specific growth rate in fermentation process, fermenting at present
Journey, thalline is sampled detection in just needing to fermentation tank in fermentation a period of time.The wriggling that feed supplement process is carried using instrument
Pump, intermittent flow feeding.
The defects of prior art, shows：(1) need by not stopping to measure the OD in fermentation process, residual sugar is fed back
Adjust feed rate；(2) manual operation requires high, time length in fermentation process；(3) microbiological contamination probability increase in fermentation process；(4)
In traditional zymotic fluid production process, feeding-system is " semistarvation " supplying technics, otherwise disposably add institute initial stage in fermentation
There is medium nutrient content, be allowed to gradually use up；The fermentation middle and later periods is manual, intermittent feed supplement, culture medium is increased
Overall ecological environment fluctuation, the loss of fermentation yield are big.
The content of the invention
The present invention provides a kind of feeding medium during fermentation Optimal Control System and method, it is possible to increase feeding medium during fermentation automation process is simultaneously
And be conducive to improve thalline fermentation density, improve destination protein expression.
According to the first aspect of the invention, the present invention provides a kind of feeding medium during fermentation Optimal Control System, which includes hair
Fermentation tank, feed supplement bottle, the glucose electrode for monitoring in above-mentioned fermentation tank sugared concentration in real time and for monitoring above-mentioned fermentation in real time
The optical density electrode of thalline OD in tank, and for be pumped into supplemented medium external feed supplement pump and external computerized control system,
It connects above-mentioned glucose electrode, optical density electrode and feed supplement pump, grows speed according to above-mentioned sugared concentration, thalline OD and the ratio of setting
Rate controls the speed of service of above-mentioned feed supplement pump, from above-mentioned benefit in the form of the fed-batch mode and continuous stream for controlling specific growth rate add
Expect that bottle supplements supplemented medium to above-mentioned fermentation tank.
Further, the feed rate of above-mentioned feed supplement pump is drawn according to equation below：
ωv=(eμ·OD-OD)·V/c/V0；
Wherein, ωvRepresent the speed of service of feed supplement pump relative to the percentage of its full speed；μ represents the ratio growth speed of setting
Rate, the i.e. increased biomass of thalline institute of unit mass per hour；V represents fermentation volume；C represents glucose in supplemented medium
Concentration；V0Represent the feeding volume of feed supplement pump full speed running in the unit interval.
Further, said system is additionally included in the tails assay mass spectrograph of above-mentioned fermentation tank tail gas end addition, for surveying
Measure oxygen uptake rate (OUR), carbon dioxide evolution rate (CER) and respiratory quotient (RQ).
Further, said system further includes the movement by thalli growth situation in network real time and on line monitoring fermentation tank
Terminal.
According to the second aspect of the invention, the present invention provides a kind of feeding medium during fermentation optimal control method, and this method includes：Make
Sugared concentration in fermentation tank is monitored in real time with the glucose electrode for being connected to fermentation tank, and fermentation tank is monitored in real time using optical density electrode
Middle thalline OD；The external computerized control system pumped using above-mentioned glucose electrode, optical density electrode and feed supplement is connected to, according to upper
The specific growth rate for stating sugared concentration, thalline OD and setting controls the speed of service of above-mentioned feed supplement pump, to control specific growth rate
The form that fed-batch mode and continuous stream add supplements supplemented medium from feed supplement bottle to above-mentioned fermentation tank.
Further, the feed rate of above-mentioned feed supplement pump is drawn according to equation below：
ωv=(eμ·OD-OD)·V/c/V0；
Wherein, ωvRepresent the speed of service of feed supplement pump relative to the percentage of its full speed；μ represents the ratio growth speed of setting
Rate；V represents fermentation volume；C represents the concentration of glucose in supplemented medium；V0Represent feed supplement pump full speed running in the unit interval
Feeding volume.
Further, the above method is additionally included in above-mentioned fermentation tank tail gas end addition tails assay mass spectrograph, for measuring
Oxygen uptake rate (OUR), carbon dioxide evolution rate (CER) and respiratory quotient (RQ).
Further, the above method further includes：Pass through thalline in network real time and on line monitoring fermentation tank using mobile terminal
Upgrowth situation.
Further, the above method further includes：Control initial medium nutritional amt, when nutrient consumption totally after
Exactly cell needs feed supplement, and OD reaches induction period.
Further, the above method according to every 1 gram of sugar make 1 liter of bacterium solution increase by the estimated specific growth rate of standard of 1 OD with
Next step feed rate is deduced, and is automatically adjusted by computer.
The present invention realizes the continuous addition of supplemented medium by adding carbon source, nitrogen source, inorganic salts and trace element.
The feeding medium during fermentation Optimal Control System and method of the present invention, is pumped using external feed supplement, to control specific growth rate
The form that fed-batch mode and continuous stream add carries out feed supplement, and thalline can be controlled to express egg in Induction period with certain growth rate
In vain, ensure that protein normal folds, expressing quantity can be effectively improved, expressing quantity improves 30%, and cell concentration improves 35%.
Feeding medium during fermentation is carried out using the system and method for the present invention, operating process is easy, and sampling number is few, and pollution probability is reduced.
Brief description of the drawings
Fig. 1 is the structure diagram of the feeding medium during fermentation Optimal Control System of one embodiment of this invention；
The fermented liquid supernatant and the egg of full bacterium that Fig. 2 obtains for the fermentation process of one embodiment of the invention and a comparative example
White yield result figure, wherein supernatant 1 and full bacterium 1 represent being pumped as a result, carrying feed supplement using fermentation tank for comparative example, and use is semicontinuous
The mode that stream adds, feeding strategy is by sampling and measuring residual sugar, and fed-batch test adds glucose after residual sugar runs out of；2 He of supernatant
Full bacterium 2 represent embodiment as a result, compared to supernatant 1 obtained by comparative example and the output increased of full bacterium 1 30%.
Embodiment
The present invention is described in further detail below by embodiment combination attached drawing.
As shown in Figure 1, the feeding medium during fermentation Optimal Control System of one embodiment of the invention includes：Fermentation tank 1, feed supplement
Bottle 2, glucose electrode 3, optical density electrode 4, external feed supplement pump 5 and external computerized control system 6, wherein glucose electrode
3 are used to monitor sugared concentration in fermentation tank 1 in real time, and optical density electrode 4 is used to monitor thalline OD in fermentation tank 1, external benefit in real time
Material pump 5 is used to be pumped into supplemented medium, external computerized control system 6 connection glucose electrode 3, optical density electrode 4 and feed supplement
Pump 5, for controlling the speed of service of feed supplement pump 5 according to the specific growth rate of sugared concentration, thalline OD and setting, so that fermentation
Optimization of material makeup control system is in the form of the fed-batch mode and continuous stream for controlling specific growth rate add from feed supplement bottle 2 to fermentation tank 1
Supplement supplemented medium.
In one embodiment of the invention, feed supplement pump 5 is peristaltic pump (such as Lange board peristaltic pump).Peristaltic pump it is initial
The speed of service is the 1% of peristaltic pump full speed.
In one embodiment of the invention, the feed rate of feed supplement pump is drawn according to equation below：
ωv=(eμ·OD-OD)·V/c/V0；
Wherein, ωvRepresent the speed of service of feed supplement pump relative to the percentage of its full speed；μ represents the ratio growth speed of setting
Rate；V represents fermentation volume；C represents the concentration of glucose in supplemented medium；V0Represent feed supplement pump full speed running in the unit interval
Feeding volume.
Technical scheme establishes a kind of fermentation model and matches corresponding fed-batch mode, and combines computer, reaches
To automatic feedback control fed-batch fermentation；Using the external feed supplement pump (such as peristaltic pump) of fermentation tank, using glucose electrode, optical density electricity
Pole and external computerized control system；Fed-batch mode is to control specific growth rate (the increased bacterium of thalline institute of unit mass per hour
The scale of construction), control thalline, with certain growth rate expressing protein, ensures that albumen can be folded normally in induction period；Using external benefit
Material pump, continuous stream add, and ensure that fermentation process thalli growth is steady；Realized by adding carbon source, nitrogen source, inorganic salts and trace element
The continuous addition of supplemented medium.
In addition, technical scheme can also include the following technological means：Tail is added at fermentation tank tail gas end
Qi leel analysis mass spectrograph is used to measure OUR, CER and RQ, feeds back thalli growth situation；The nutritional amt of initial medium is controlled, is being waited
To nutrient consumption, totally exactly cell needs feed supplement afterwards, and OD reaches induction period；Make 1 liter of bacterium solution according to every 1 gram of sugar
Increase by the estimated specific growth rate of standard of 1 OD to deduce next step feed rate, and automatically adjusted by computer；And make
Pass through thalli growth situation in network real time and on line monitoring fermentation tank with mobile terminal.
Embodiment 1
By taking the 30L rosseta bacterial strains that ferment as an example.
(1) first cell culture medium：Sodium chloride 10g/L, tryptone 10g/L, yeast extract 5g/L.
(2) secondary medium：Tryptone 12g/L, yeast extract 24g/L, three water dipotassium hydrogen phosphate 16.45g/L, phosphoric acid
Potassium dihydrogen 2.31g/L, glycerine 5g/L.
(3) fermentation medium includes：48mmol/L disodium hydrogen phosphates, 20mmol/L sodium dihydrogen phosphates, 16mmol/L lemons
Sour ammonium, the yeast extract of the glucose and 1.9g/L of 9mmol/L epsom salts and 24g/L, 3.8g/L tryptones are micro
Element 0.1%, 0.09% defoamer, pH 7.2.Wherein epsom salt, glucose and trace element individually sterilizing.
(4) supplemented medium includes：The yeast of 80mmol/L epsom salts, the glucose of 640g/L and 38.8g/L
Soak powder and 19.4g/L tryptones, the defoamer of trace element 0.15% and 0.09%.Wherein yeast extract, tryptone and
Defoamer individually sterilizes.
(5) single bacterium colony on tablet is chosen in first cell culture medium, first order seed culture to OD is 4.0, is inoculated into 1%
In secondary medium, treat that OD reaches 2.5 and is inoculated into fermentation tank with 3% ratio volume ratio.
(6) fermentation temperature is arranged to 37 DEG C, and pH is not controlled, and associates dissolved oxygen and rotating speed, and dissolved oxygen control is trained more than 50%
Support, treat that dissolved oxygen starts to rebound, about 7h, OD is about 20 at this time, carries out feed supplement, and start to be cooled to 30 DEG C.
(7) feed supplement uses Lange board peristaltic pump, and initial feed rate is the 1% of Lange board peristaltic pump full speed, is controlled at this time
pH6.5。
(8) monitored in real time by optical density electrode and glucose electrode, by sugared concentration, thalline OD, the ratio of setting is grown
Speed 0.11, fermentation volume, the full speed feed rate (L/h) of feed supplement pump, several parameters such as sugared concentration (640g/L) of feed supplement, are pressed
According to formula ω v=(e0.11*OD-OD) * 30/0.64/3240, the feed rate of pump is calculated, control residual sugar content is not less than
0.5g/L, if remaining sugar concentration is more than 0.5g/L, the speed of feed supplement hour by more than carries out.PH is controlled with hand per hour
The dynamic value by pH controls improves 0.03, remains unchanged after start bit 6.5, most Zhongdao 7.0.Dissolved oxygen and rotating speed are associated, by dissolved oxygen control
System is 50%.During observe thalline oxygen uptake rate and respiratory quotient.Method induces 15 hours accordingly, and OD is up to 80.
Note：Formula ω v=(e0.11* OD-OD) in * 30/0.64/3240, ω represent the speed of service of feed supplement pump relative to
The percentage of full speed；0.11 represents the specific growth rate of control；30 represent fermentation volume 30L；0.64 represents 1mL supplemented mediums
Middle glucose content 0.64g；3240 represent that the feeding volume that feed supplement in one hour pumps full speed is 3240mL.
Comparative example 1
(1) first cell culture medium：Sodium chloride 10g/L, tryptone 10g/L, yeast extract 5g/L.
(2) secondary medium：Tryptone 12g/L, yeast extract 24g/L, three water dipotassium hydrogen phosphate 16.45g/L, phosphoric acid
Potassium dihydrogen 2.31g/L, glycerine 5g/L.
(3) fermentation medium includes：48mmol/L disodium hydrogen phosphates, 20mmol/L sodium dihydrogen phosphates, 16mmol/L lemons
Sour ammonium, the yeast extract of the glucose and 1.9g/L of 9mmol/L epsom salts and 24g/L, 3.8g/L tryptones are micro
Element 0.1%, 0.09% defoamer, pH 7.2.Wherein epsom salt, glucose and trace element individually sterilizing.
(4) supplemented medium includes：The yeast of 80mmol/L epsom salts, the glucose of 640g/L and 38.8g/L
Soak powder and 19.4g/L tryptones, the defoamer of trace element 0.15% and 0.09%.Wherein yeast extract, tryptone and
Defoamer individually sterilizes.
(5) single bacterium colony on tablet is chosen in first cell culture medium, first order seed culture to OD is 4.0, is inoculated into 1%
In secondary medium, treat that OD reaches 2.5 and is inoculated into fermentation tank with 3% ratio volume ratio.
(6) fermentation temperature is arranged to 37 DEG C, and pH controls are 7.0, associate dissolved oxygen and rotating speed, by dissolved oxygen control more than 50%
Culture, treats that dissolved oxygen starts to rebound, about 7h, and OD is surveyed in sampling at this time.Feed supplement is carried out, feed supplement is pumped using the feed supplement of fermentation tank, and is opened
Beginning is cooled to 30 DEG C.
(7) feed supplement, offline sampling and measuring OD, measures residual sugar, when the amount of residual sugar is less than 1g/L, with constant speed feed supplement.Use
Be fermentation tank carry feed supplement pump.To mend 3 seconds, the mode stopped 107 seconds carries out feed supplement.
The Comparative result of final feeding medium during fermentation method is as shown in table 1.
Table 1
Existing feeding medium during fermentation method be using fermentation tank carry feed supplement pump, using fed-batch test plus by the way of.Feed supplement plan
Slightly it is by sampling and measuring residual sugar, fed-batch test adds glucose after residual sugar runs out of.On in protein yield electrophoresis result such as Fig. 2
Clear 1 and full bacterium 1 shown in；Feed process of the present invention, pumps (peristaltic pump), to control the feed supplement mould of specific growth rate using external feed supplement
The form that formula and continuous stream add carries out feed supplement, and obtained protein yield electrophoresis result is as shown in supernatant 2 in Fig. 2 and full bacterium 2, than existing
The output increased 30% of supernatant 1 and full bacterium 1 obtained by some feeding medium during fermentation methods.
Above content is to combine specific embodiment further description made for the present invention, it is impossible to assert this hair
Bright specific implementation is confined to these explanations.For general technical staff of the technical field of the invention, do not taking off
On the premise of from present inventive concept, some simple deduction or replace can also be made, should all be considered as belonging to the protection of the present invention
Scope.
Claims (10)
1. a kind of feeding medium during fermentation Optimal Control System, it is characterised in that the system comprises fermentation tank, feed supplement bottle, for real-time
Monitor the glucose electrode of sugared concentration and the optical density electricity for monitoring thalline OD in the fermentation tank in real time in the fermentation tank
Pole, and external feed supplement for being pumped into supplemented medium pumps and external computerized control system, its connect described glucose electrode,
Optical density electrode and feed supplement pump, the fortune for controlling the feed supplement to pump according to the sugared concentration, thalline OD and the specific growth rate of setting
Scanning frequency degree, is supplemented in the form of the fed-batch mode and continuous stream for controlling specific growth rate add from the feed supplement bottle to the fermentation tank
Supplemented medium.
2. feeding medium during fermentation Optimal Control System according to claim 1, it is characterised in that the feed rate of the feed supplement pump
Drawn according to equation below：
ωv=(eμ·OD-OD)·V/c/V0；
Wherein, ωvRepresent the speed of service of feed supplement pump relative to the percentage of its full speed；μ represents the specific growth rate of setting, i.e.,
The increased biomass of thalline institute of unit mass per hour；V represents fermentation volume；Glucose is dense in c expression supplemented mediums
Degree；V0Represent the feeding volume of feed supplement pump full speed running in the unit interval.
3. feeding medium during fermentation Optimal Control System according to claim 1, it is characterised in that the system also includes described
The tails assay mass spectrograph of fermentation tank tail gas end addition, for measuring oxygen uptake rate, carbon dioxide evolution rate and respiratory quotient.
4. feeding medium during fermentation Optimal Control System according to claim 1, it is characterised in that the system also includes pass through net
The mobile terminal of thalli growth situation in network real time and on line monitoring fermentation tank.
A kind of 5. feeding medium during fermentation optimal control method, it is characterised in that the described method includes：Use the grape for being connected to fermentation tank
Sugared electrode monitors sugared concentration in fermentation tank in real time, and thalline OD in fermentation tank is monitored in real time using optical density electrode；Using being connected to
The external computerized control system of the glucose electrode, optical density electrode and feed supplement pump, according to sugar concentration, the thalline OD and sets
Fixed specific growth rate controls the speed of service of the feed supplement pump, to control the fed-batch mode of specific growth rate and continuous stream adds
Form supplements supplemented medium from feed supplement bottle to the fermentation tank.
6. feeding medium during fermentation optimal control method according to claim 5, it is characterised in that the feed rate of the feed supplement pump
Drawn according to equation below：
ωv=(eμ·OD-OD)·V/c/V0；
Wherein, ωvRepresent the speed of service of feed supplement pump relative to the percentage of its full speed；μ represents the specific growth rate of setting；V tables
Show fermentation volume；C represents the concentration of glucose in supplemented medium；V0Represent the feed supplement of feed supplement pump full speed running in the unit interval
Volume.
7. feeding medium during fermentation optimal control method according to claim 5, it is characterised in that the method is additionally included in described
Tails assay mass spectrograph is added at fermentation tank tail gas end, for measuring oxygen uptake rate, carbon dioxide evolution rate and respiratory quotient.
8. feeding medium during fermentation optimal control method according to claim 5, it is characterised in that the method further includes：Use
Mobile terminal passes through thalli growth situation in network real time and on line monitoring fermentation tank.
9. feeding medium during fermentation optimal control method according to claim 5, it is characterised in that the method further includes：Control
The nutritional amt of initial medium, is being exactly totally that cell needs feed supplement afterwards when nutrient consumption, and OD reaches induction rank
Section.
10. feeding medium during fermentation optimal control method according to claim 5, it is characterised in that the method is according to every 1 gram of sugar
The standard that 1 liter of bacterium solution increases by 1 OD is expected specific growth rate to deduce next step feed rate, and adjusted automatically by computer
Section.
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| CN108546637A (en) * | 2018-05-23 | 2018-09-18 | 华东理工大学 | Restructured Pichia pastoris in expression glucose oxidase installation for fermenting and fermentation culture method |
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| CN111893145A (en) * | 2020-08-13 | 2020-11-06 | 山东寿光巨能金玉米开发有限公司 | Novel intelligent lysine biological fermentation method |
| CN112899125A (en) * | 2021-04-02 | 2021-06-04 | 西安交通大学 | Microalgae efficient carbon sequestration device and nutrient supplement control method |
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| CN113604523A (en) * | 2021-08-23 | 2021-11-05 | 华东理工大学 | Method for producing sophorolipid through efficient fermentation based on data mechanism fusion model |
| CN116153388A (en) * | 2023-04-19 | 2023-05-23 | 北京蓝晶微生物科技有限公司 | Quantitative relation model and application, cell state monitoring method, device and system |
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