CN1231592C - D-ribose fermentation process regulating method - Google Patents
D-ribose fermentation process regulating method Download PDFInfo
- Publication number
- CN1231592C CN1231592C CN 02156496 CN02156496A CN1231592C CN 1231592 C CN1231592 C CN 1231592C CN 02156496 CN02156496 CN 02156496 CN 02156496 A CN02156496 A CN 02156496A CN 1231592 C CN1231592 C CN 1231592C
- Authority
- CN
- China
- Prior art keywords
- fermentation
- control
- ribose
- glucose
- sugar
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention relates to a majorized regulation and control method in a D-ribose fermentation process, which is a fermentation control method taking controlling dissolved oxygen and pH value as the main line and controlling key variables, such as the thallus growth, the wear rate of substrates, the product generating rate, etc. as the main contents and supplement sugar by fed-batch in real time. A fermentation result in a fermenting tank of which the capacity volume is 5000l indicates that the D-ribose sugar producing level can be enhanced by 20 to 34%, the fermentation period is shortened to 42 to 50 hr, and the fill factor of the fermenting tank is increased to more than 80% from 70% by using the method. Moreover, the fermentation process is easy to control, and the repeatability is strengthened.
Description
(1) technical field
The present invention relates to a kind of regulate and control method of D-ribose fermenting process.
(2) background technology
The preparation method of D-ribose comprises methods such as extraction separation, chemosynthesis and microbial fermentation from crude substance.Extraction separation D-ribose from natural goods, leaching process is numerous and diverse, manufacturing cost is high, is not suitable for large-scale production.With regard to synthesis method, people have successively attempted from D-pectinose (Gehrke and Aichner, 1927), maltonic acid (Steiger, 1936), D-glucose (Korczynski A et al.1979), L-L-glutamic acid and D-wood sugar (Lacourt Gadras, et al.1992) synthetic D-ribose, before 20th century the eighties, the main both at home and abroad suitability for industrialized production of carrying out D-ribose by glucose chemistry synthesis method that adopts, promptly by glucose through oxidation, displacement, transform, displacement again, acidifying, lactonize, seven steps reaction such as reduction makes D-ribose, this method is the step complexity not only, equipment used is many, yield is low, the manufacturing cost height, and its related tribute electrode reduction reaction also can cause serious environmental to pollute.After entering 20th century the nineties, people bring into use Production by Microorganism Fermentation D-ribose, fermentation method carries out at normal temperatures and pressures, the starting material wide material sources, avoided the environmental pollution of chemical synthesis again, being called as the best approach of producing D-ribose, also is the method with very big suitability for industrialized production potentiality.
The fermentative Production process is very complicated a chemical transformation and a physiological change combined process, and improving fermentation level has two approach, the one, filter out good bacterial classification, and the 2nd, draw the optimal culture condition, the control device that are complementary with the purpose bacterial strain.The former is the strain improvement technology that is based upon in the metabolic control fermentation research, and the latter is the fermentation control techniques that is based upon on the biochemical reaction engineering basis.Have only both closely in conjunction with could finally realizing high-caliber fermentative production.
For a long time, people are placed on focus in the seed selection of superior strain morely, about the research that fermenting process is optimized and controls less.Required some are cultivated control conditions, technical indicator and are often obtained shaking under bottle condition in the actual production, and this low-level research method has objectively restricted the further performance of bacterial classification potentiality.Moreover, owing to shaking under bottle condition and can't set up organic contact between aspect purpose microorganism physio-biochemical characteristics, nutrition supply and operation, the fermentation equipment three, when the reactor scale changed, fermentation was the result usually can not repeat.And the also basic place that reduces along with production-scale expansion (producing to big to pilot scale from shaking bottle) process fermentation level of some leavened prod just of this point.
Fermenting process is optimized the purpose of control, is to obtain the highest production concentration, the highest production intensity (being generally the amount that every liter fermented liquid per hour is converted into product, often is unit with g/Lh) and the highest transformation efficiency simultaneously.And when attempting to obtain higher production concentration, must mean the further decline of transformation efficiency and production intensity with batch fermentation.Add the fermentation Research of optimization about fermentation parameter optimal control and stream in the fermentation industry in recent years, for the production level that improves batch fermentation provides possible approach.At certain first sugared concentration bottom fermentation, under fermentation parameter optimal control condition, improve total sugar concentration by fed-batch mode, and the sugared concentration of keeping fermenting process is in suitable level, just might obtains high transformation efficiency, high production intensity and high production concentration simultaneously.
D-ribose is fermented, people's sight focuses mostly at the screening of strain excellent, the aspects such as improvement of culture medium prescription for many years, the Comprehensive Control of the fermentation parameter (as dissolved oxygen, air flow and pH value, tank pressure and mixing speed) that is even more important for seeming by the D-ribose fermentation that the oxidative phosphorylation approach could form, in document in the past, just mention very faintly, or just certain parameter is wherein reported.As the control of pH in the fermenting process, mostly be and only control the initial pH value of fermention medium or only mention certain optimal ph (Asain and Kono 1984; Asain et al1978; Sasjima and Yoneda 1989), and the pH of whole fermentation process is controlled, do not see argumentation; And for example set forth and controlled the flat accumulation that helps D-ribose of water containing high dissolved oxygen during the fermentation, but do not disclosed the demand (P.De Wulf E.J.Vandamme, J ApplMicrobiol 1997) of each stage of fermentation oxygen; The fermentation period of existing ribose superior strain is long, generally at 60～90 hours, have in addition surpassed 100 hours (P.De Wulf E.J.Vandamme, ApplMicrobiol Biotechnol 1997,48:141～148), see Table one.Although the D-ribose content that some bacterial strain accumulates in fermention medium is up to more than the 90g/L, because to not effectively control of fermenting process, most fermentation periods reached more than 72 hours, even 144 hours (seeing Table 1), its production intensity can not be satisfactory.
Table one: the ribose fermentation period of different strains
The strain fermentation cycle (hour)
B.subtilis?ATCC?21360 68
B.pumilus ATCC?21357 55
B.subtilis?ATCC?21951 144
B.subtilis?ATCC?21951 90
B.subtilis?ATCC?21952 72
B.subtilis?ATCC?31092 60
B.pumilus ATCC?91094 60
Bacillus?sp.EMP-58 55
B.subtilis?ATCC?21951 72
B.subtilis?ATCC?21951 72
B.subtilis?IFO?1538 72
B.subtilis?ATCC?21951 110
B.subtilis?ATCC?21951 156
B.subtilis?ATCC?21951 84
In the D-ribose fermentative production method, the microorganisms of adopting are the bacterial strain of bacillus more at present, wherein to use the bacterial strain that subtilis belongs to and bacillus pumilus belongs in the majority.And sophisticated method does not appear as yet about the optimal control of these bacterial strains D-ribose fermenting process.
The fermentation of D-ribose is the subject of a youth, and its fermenting process is optimized Control Study, determines the optimal control condition that it is best, is that fermentation industry realizes the prerequisite that efficient stable is produced.
(3) summary of the invention
[problem that will solve]
The purpose of this invention is to provide a kind of effective regulate and control method that utilizes the D-ribose fermenting process that bacillus micro-organism carries out.
[technical scheme]
The factor that influences the fermentation of D-ribose can be divided into two classes: a class is the environmental parameter of fermenting process, and as temperature, tank pressure, mixing speed, pH value, dissolved oxygen etc., another kind of is key variables, forms speed etc. as thalli growth amount, base consumption speed, product.
The present invention adopts advanced fermenting process distributed control system-FPC type distributed control system (DCS), first kind environmental parameter is carried out on-line Control and monitoring, and measure the mode of the fermenting process second class key variables in conjunction with timing sampling, systematic study is carried out in control to the ribose fermenting process, the optimum control condition of each parameter of definite kernel sugar-fermenting different steps, convection current adds the control condition of mending sugar and explores on this basis, determined a kind of with dissolved oxygen, the pH value is controlled to be main line, various parameters in the Comprehensive Control fermenting process, and stream adds effective regulate and control method of mending sugar in good time.
More particularly, the regulate and control method of D-ribose fermenting process of the present invention has following feature:
With culture temperature, mixing speed, the tank pressure of fermenting process distributed control system-FPC type distributed control system (DCS) on-line Control D-ribose fermentation and pH value, the dissolved oxygen amount of the different steps of fermenting, and timing sampling is measured somatic cells concentration, concentration of substrate and product growing amount in the fermenting process, stream adds and mends sugar when sugared concentration subcritical value, keeps fermenting process to be in optimum regime all the time.
This method is a kind ofly to serve as theme with control dissolved oxygen, pH value, is main contents with key variables such as control thalli growth amount, base consumption speed and product generating rates, and stream adds and mends sugared fermentation controlling method in good time.
This method is determined and the maintenance optimal conditions of fermentation by using other parameter (as culture temperature, tank pressure, mixing speed) in fermenting process distributed control system-online Comprehensive Control dissolved oxygen of FPC type distributed control system (DCS), pH value and the fermenting process; By measuring somatic cells concentration, concentration of substrate and product growing amount, obtain the Changing Pattern of key variables such as biomass growth rate in the fermenting process, base consumption speed and product generating rate, and, determine that just sugared concentration, stream add benefit sugar opportunity and mends sugared mode in best fermentation in conjunction with different first sugared concentration fermentation test results.On this basis, with optimal conditions of fermentation, best fermentation just sugared concentration is fermented, and mends sugar (glucose) during the fermentation in good time, finishes the ribose fermenting process of optimal control thus.
More than said fermenting process comprise the process of using various microbial fermentations to prepare D-ribose, wherein preferably use the process of the fermentative preparation D-ribose of Bacillus strain.
In aforesaid method, the culture temperature in the wherein said fermenting process is in 36～38 ℃ of scopes, and tank pressure is at 0.5～0.8kg/cm
2In the scope, mixing speed is in 180～220 rev/mins of scopes.
In aforesaid method, the optimal ph scope of wherein said fermentation different steps is respectively: earlier fermentation (0～18 hour) 6.50～7.0; Fermentation middle and later periods (after 18 hours) 6.00～6.50.
In aforesaid method, the best dissolved oxygen scope of wherein said fermentation different steps is respectively: earlier fermentation (0～18 hour) 18～30%; Fermentation middle and later periods (after 18 hours) 20～35%.
In aforesaid method, wherein the threshold value of said sugared concentration is 2.5%, stream add the mode of mending sugar be with the 100-130 liter/hour flow velocity stream add glucose solution, make the glucose content in the fermention medium constant in the scope of 2.5%-3.0%, the concentration of the glucose solution that institute's stream adds is 30-50%, and volume is the 10-20% of fermentor tank volume.
Fermenting process control comprises steady operation conditions and optimizes two aspects of fermentation parameter, the former can realize by conventional regulating loop, and the optimal control of system needs the Changing Pattern according to the key variables of fermenting process, jar temperature, tank pressure, pH, dissolved oxygen etc. are carried out optimizing calculate, to determine the optimal control policy of these variablees.To this, adopt conventional instrument to realize.Have simple, reliable, high performance characteristics and adopt distributed control system to carry out the automatic control of biological process.
The FPC type distributed control system of using in the method for the present invention (DCS) (as shown in Figure 1) is made of upper computer and intelligence instrument, remote I/O module, gets in touch by process bus.Control software is the configuration control software of developing with VISUAL BASIC6.0 under the WIN98 environment.Use therein upper computer is an industrial computer, and housing is directly controlled by intelligence instrument.
The function of intelligence instrument be become give, digital display, PID be from the control of adjusting, hand behaviour, no disturbance switching, RS485 communication; The function of remote I/O module is A/D conversion, I/O input and output, RS485 communication; Intelligence instrument and remote I/O module are mainly finished each parameter The real time measure and control.By measuring transducer fermenting process stir current, jar temperature, tank pressure, charge flow rate, pH, dissolved oxygen, foam etc. are measured in real time, all data are sent upper computer analysis, storage by fieldbus.Upper computer requires to carry out stepless control by set(ting)value according to control simultaneously, the major control amount has: jar temperature, tank pressure, charge flow rate, pH, dissolved oxygen, froth breaking, feed supplement etc., and by analyzing regularly storage, and in time on indicating meter, show, report to the police if having unusually immediately, realization is carried out comprehensive monitoring and closed-loop control to the fermenting process environmental parameter, and the reproducibility of fermentative production is improved.
FPC type distributed control system (DCS) has following function:
(1), data acquisition and processing (DAP)
To fermentation parameters such as jar temperature, tank pressure, charge flow rate, pH, dissolved oxygen, froth breaking, feed supplements, realize reliable on-line measurement, and, in time on indicating meter, show by analyzing regularly storage, report to the police if having unusually immediately, realize the fermenting process environmental parameter is carried out comprehensive monitoring and closed-loop control.
(2), fermenting process sequential control
Finish the open and close of respective valves such as fermenting process sterilization, inoculation culture, input and output material by logic function, operations such as the start and stop of agitator motor make it not only safe but also lower labour intensity, thereby improve serviceability.
(3), direct digital control and set(ting)value control
Can realize various feedback control (as PID control, serial connection control, adaptive control etc.) easily to parameters such as jar temperature, tank pressure, charge flow rate, pH values, and can require to go to change set(ting)value according to Optimizing operation.
(4), optimal control
By the optimization of jar temperature, the optimization of pH value and the optimization of oxygen transmittance process etc., determine the condition of fermentation optimization control, realize the controllability operation of fermentative production, improve fermentation production rate, shorten fermentation time.
The main instrument selection of this system is as follows:
Temperature: waterproof type platinum resistance thermometer sensor, Pt100;
Pressure: capacitance pressure transmitter, diaphragm pressure gage;
Flow: whirling current transmitter (antidetonation type), glass ball float under meter;
Foam: capacitance type digital material-level switch;
PH: high temperature resistant pH value measuring system (converter that Switzerland Mettler Toledo company transmitter, Singapore EUTECH INSTRUMENTS company produce)
Dissolved oxygen: high temperature resistant dissolved oxygen measuring system (converter that Switzerland Mettler Toledo company transmitter, Singapore EUTECH INSTRUMENTS company produce)
Stir current: current transformer
Control instruments: the small intelligent regulation meter of band RS485 communication
Variable valve: the magnetic valve of pneumatic diaphragm control valve, corrosion-and high-temp-resistant
Regulation according to patent law of china and its detailed rules for the implementation, bacillus pumilus P8069 of the present invention and subtilis S6023 are deposited in China Microbial Culture Preservation Commission common micro-organisms center (CGMCC) on December 5th, 2002, and preserving number is respectively CGMCC 0845 and CGMCC 0846.
[beneficial effect]
By optimal control to D-ribose fermenting process, comprise the main line that is controlled to be with dissolved oxygen and pH value, the various parameters of fermenting process are carried out comprehensive parameters, and mended control method such as sugar in good time, D-ribose produces sugar level can improve 20～34%, fermentation period foreshortens to 42～50 hours, the packing factor of fermentor tank is increased to more than 80% by 70%, and fermenting process is easy to control, repeatability strengthens.
(4) description of drawings
Fig. 1 is a distributed control system overall construction drawing of the present invention.
(5) embodiment
Below further specify the present invention with non-limiting example.Below the glucose that uses among each embodiment contain 1 molecular crystal water, pure content is 90%.
Embodiment 1
Use bacillus pumilus (B.pumilus) CGMCC 0845.By sorbyl alcohol 2.0% (w/w), corn steep liquor 2.0%, yeast extract paste 0.1%, dipotassium hydrogen phosphate 0.3%, potassium primary phosphate 0.1%, in the 350L seed culture medium (pH 7.0) that sal epsom 0.05% is formed, control seeding tank tank pressure 1.0kg/cm
2200 rev/mins of rotating speeds, 36 ℃ of culture temperature, cultivate and insert after 16 hours by glucose 16.0% (w/w), corn steep liquor 2.6%, yeast extract paste 0.1%, ammonium sulfate 0.7%, manganous sulfate 0.005%, the control mixing speed is 180 rev/mins in the 3200L fermention medium (pH 7.0) that lime carbonate 2.2% is formed, 36 ℃ of culture temperature.Earlier fermentation 0～18 hour, be controlled at pH 6.5～7.0, dissolved oxygen 20～25%; The fermentation middle and later periods is controlled at pH 6.3～6.5, dissolved oxygen 25～30%.Fermented preceding 18 hours, glucose consumption is slow, mainly is the thalli growth stage.Ferment after 18 hours, glucose consumption begins to accelerate, and D-ribose begins accumulation, and thalli growth speed slightly descends.During by 24 hours, thalli growth enters stationary phase, and the thalli growth amount reaches 15g wet thallus/100ml fermented liquid.Quickening along with glucose consumption speed, product accumulation enters the quick product sugar phase, and the sugar consumption speed in this stage per hour is about 0.6～0.8% very soon, product forms and to enter the peak period, and transformation efficiency can reach 50～55% (weight transformation efficiencys) sometimes even reach more than 60% approximately.Fermentation culture 32 hours, the glucose in the fermented liquid reduced to 2.5～3.0%, and at this moment, the sugared concentration in the control fermented liquid begins stream with 120 liters of/hour flow velocitys and adds and mend sugar in 2.5～3.0% scope, and the sugared concentration that stream adds is 40%, volume 550L.Stream is kept above-mentioned control condition after adding the end of benefit sugar, continues to cultivate 45 hours, and glucose exhausts, pH value and the rising of dissolved oxygen straight line, and fermentation ends, the content of D-ribose is 90.60g/L in the fermented liquid, fermentating liquid volume is 4050L.
Comparing embodiment 1-1 (do not mend sugar, control condition is identical, and first sugared concentration improves)
Use bacillus pumilus CGMCC 0845.With forming the 350L seed culture medium identical, that culture condition is identical with embodiment 1 substratum, cultivate after 18 hours for 36 ℃, access is by glucose 21.88% (w/v), corn steep liquor 2.5%, dry yeast 0.1%, ammonium sulfate 0.6%, manganous sulfate 0.005% is in the 3200L fermention medium (pH 7.0) that lime carbonate 2.2% is formed.Fermentation each parameter control condition of different steps and embodiment 1 are identical, fermentation culture 45 hours, and the content of D-ribose is 73.65g/L in the fermented liquid, fermentating liquid volume is 3500L.
Comparing embodiment 1-2 (do not mend sugar, control condition is simplified, and first sugared concentration improves)
Use bacillus pumilus CGMCC 0845.With forming the 350L seed culture medium identical, that culture condition is identical with embodiment 1 substratum, cultivate after 18 hours for 36 ℃, access is by glucose 21.88% (w/v), corn steep liquor 2.5%, dry yeast 0.1%, ammonium sulfate 0.6%, manganous sulfate 0.005% is in the 3200L fermention medium (pH 7.0) that lime carbonate 2.2% is formed.180 rev/mins of control mixing speed, 36 ℃ of culture temperature by 1: 0.9 ventilation, were cultivated 52 hours 36 ℃ of following aeration-agitations, and the content of D-ribose is 69.60g/L in the fermented liquid, and fermentating liquid volume is 3500L.
Embodiment 2
Use subtilis (B.subtilis) CGMCC 0846.By glucose 2.0% (w/w), yeast extract paste 0.3%, corn steep liquor 0.05%, dipotassium hydrogen phosphate 0.3%, potassium primary phosphate 0.1%, in the 350L seed culture medium that sal epsom 0.05% (pH 7.0) is formed, control seeding tank tank pressure 1.0kg/cm
2, 200 rev/mins of rotating speeds, 36 ℃ of culture temperature were cultivated after 18 hours, insert by glucose 16.0% (w/w), corn steep liquor 2.6%, ammonium sulfate 0.7%, manganous sulfate 0.005% is in the 3200L fermention medium (pH 7.0) that lime carbonate 2.2% is formed.180 rev/mins of control mixing speed, 36 ℃ of culture temperature.Earlier fermentation 0～18 hour, be controlled at pH 6.5～7.0, dissolved oxygen 18～22%; Fermentation the middle and later periods be controlled at pH 6.0～6.2, dissolved oxygen 20～25%; At 36 ℃, 0.60kg/m
2Following aeration-agitation cultivate.Fermented preceding 18 hours, glucose consumption is slow, mainly is the thalli growth stage.Ferment after 18 hours, glucose consumption begins to accelerate, and D-ribose begins accumulation, and thalli growth speed slightly descends.During by 24 hours, thalli growth enters stationary phase, and the thalli growth amount reaches 16g wet thallus/100ml fermented liquid.Quickening along with glucose consumption speed, product accumulation enters the quick product sugar phase, and the sugar consumption speed in this stage per hour is about 0.7～0.9% very soon, product forms and to enter the peak period, and transformation efficiency can reach 50～55% (weight transformation efficiencys) sometimes even reach more than 60% approximately.Fermentation culture 33 hours, the glucose in the fermented liquid reduced to 2.5～2.8%, and at this moment, the sugared concentration in the control fermented liquid with 120 liters/hour speed, begins stream and adds and mend sugar in 2.5～2.8% scope, and the sugared concentration that stream adds is 45%, volume 600L.After mending the sugar end, keep above-mentioned control condition, fermentation culture 46 hours, glucose exhausts, fermentation ends, the content of D-ribose is 91.35g/L in the fermented liquid at this moment, fermentating liquid volume is 4100L.
Comparing embodiment 2-1 (do not mend sugar, control condition is identical, and first sugared concentration improves)
Use subtilis CGMCC 0846.With forming the 350L seed culture medium identical, that culture condition is identical with embodiment 1 substratum, cultivate after 18 hours for 36 ℃, insert by glucose 21.88% (w/v), corn steep liquor 2.6%, ammonium sulfate 0.7%; Manganous sulfate 0.005% is in the 3200L fermention medium (pH 7.0) that lime carbonate 2.2% is formed.Each parameter control of fermentation different steps is identical with embodiment 4, fermentation culture 42 hours, and glucose exhausts fermentation ends, and the content of D-ribose is 75.60g/L in the fermented liquid at this moment, and fermentating liquid volume is 3500L.
Comparing embodiment 2-2 (do not mend sugar, control condition is simplified, and first sugared concentration improves)
Use subtilis CGMCC 0846.With forming the 350L seed culture medium identical, that culture condition is identical with embodiment 1 substratum, cultivate after 18 hours for 36 ℃, access is by glucose 21.88% (w/v), corn steep liquor 2.6%, ammonium sulfate 0.7%, manganous sulfate 0.005% is in the 3200L fermention medium (pH 7.0) that lime carbonate 2.2% is formed.180 rev/mins of control mixing speed, 36 ℃ of culture temperature by 1: 0.8 ventilation, were cultivated 56 hours 36 ℃ of following aeration-agitations, and the content of D-ribose is 68.60g/L in the fermented liquid, and fermentating liquid volume is 3500L.
Embodiment 3
Use bacillus pumilus ATCC 31093.By sorbyl alcohol 2.0%, corn steep liquor 2.0%, dipotassium hydrogen phosphate 0.3%, potassium primary phosphate 0.1%, in the 350L seed culture medium that each 100 μ g/ml (pH 7.0) of tyrosine and phenylalanine form, control seeding tank tank pressure 1.0kg/cm
2200 rev/mins of rotating speeds, 36 ℃ of culture temperature, cultivate and insert after 16 hours by glucose 16.0% (w/w), dry yeast 1.2%, ammonium sulfate 0.6%, manganous sulfate 0.005%, lime carbonate 2.2%, tryptophane, tyrosine and phenylalanine are equipped with in the 3200L fermention medium (pH 7.0) of 50 μ g/ml composition, adopt the fermentating controling condition identical with embodiment 1, fermentation culture 33 hours, the glucose in the fermented liquid reduced to 2.5～3.0%, sugared concentration in the control fermented liquid is in 2.5～3.0% scopes, begin stream with 120 liters/hour flow velocitys and add benefit sugar, the sugared concentration that stream adds is 40%, volume 550L.Fermentation culture 48 hours, glucose exhausts, and pH, dissolved oxygen straight line rise, fermentation ends, the content of D-ribose is 89.79g/L in the fermented liquid at this moment, fermentating liquid volume is 4050L.
Comparing embodiment 3-1 (do not mend sugar, control condition is identical, and first sugared concentration improves)
Use bacillus pumilus ATCC 31093.With forming the 350L seed culture medium identical, that culture condition is identical with embodiment 3 substratum, cultivate after 18 hours for 36 ℃, access is by glucose 21.88% (w/v), dry yeast 1.2%, ammonium sulfate 0.7%, manganous sulfate 0.005%, lime carbonate 2.2% is in the 3200L fermention medium (pH 7.0) that each 50 μ g/ml of tryptophane, tyrosine and phenylalanine form.Add the benefit sugar except that not carrying out stream, fermentation each parameter control condition of different steps and embodiment 1 are identical, fermentation culture 45 hours, and glucose exhausts, and pH, dissolved oxygen straight line rise fermentation ends.The content of D-ribose is 75.65g/L in the fermented liquid, and fermentating liquid volume is 3500L.
Comparing embodiment 3-2 (do not mend sugar, control condition is simplified, and first sugared concentration improves)
Use bacillus pumilus ATCC 31093.With forming the 350L seed culture medium identical, that culture condition is identical with embodiment 3 substratum, cultivate after 18 hours for 36 ℃, access is by glucose 21.88% (w/v), dry yeast 1.2%, ammonium sulfate 0.7%, manganous sulfate 0.005% is in the 3200L fermention medium (pH 7.0) that each 50 μ g/ml of lime carbonate 2.2% tryptophane, tyrosine and phenylalanine form.180 rev/mins of control mixing speed, 36 ℃ of culture temperature by 1: 0.8 ventilation, were cultivated 58 hours 36 ℃ of following aeration-agitations, and the content of D-ribose is 66.79g/L in the fermented liquid, and fermentating liquid volume is 3500L.
Embodiment 4
Use subtilis IFO 13621.With forming identical with embodiment 3 substratum, the 350L seed culture medium that culture condition is identical, 36 ℃ of culture temperature, cultivate and insert after 18 hours by glucose 16% (w/v), dry yeast 1.0%, ammonium sulfate 0.6%, manganous sulfate 0.005%, lime carbonate 2.2%, tryptophane, in the 3200L fermention medium (pH 7.0) that each 50 μ g/ml of tyrosine and phenylalanine form, adopt the fermentating controling condition identical with embodiment 2, fermentation culture 32 hours, glucose in the fermented liquid reduces to 2.5～3.0%, and the sugared concentration in the control fermented liquid begins stream with 120 liters/hour flow velocitys and adds benefit sugar in 2.5～3.0% scope, the sugared concentration that stream adds is 40%, volume 400L.Fermentation culture 48 hours, glucose exhausts, and pH, dissolved oxygen straight line rise, fermentation ends, the content of D-ribose is 85.29g/L in the fermented liquid at this moment, fermentating liquid volume is 4100L.
Comparing embodiment 4-1 (do not mend sugar, control condition is identical, and first sugared concentration improves)
Use subtilis IFO 13621.With forming the 350L seed culture medium identical, that culture condition is identical with embodiment 4 substratum, cultivate after 18 hours for 36 ℃, access is by glucose 21.88% (w/v), dry yeast 1.0%, ammonium sulfate 0.6%, manganous sulfate 0.005%, lime carbonate 2.2% is in the 3200L fermention medium (pH 7.0) that each 50 μ g/ml of tryptophane, tyrosine and phenylalanine form.Fermentation each parameter control condition of different steps and embodiment 2 are identical, fermentation culture 45 hours, and glucose exhausts, and pH, dissolved oxygen straight line rise, fermentation ends, the content of D-ribose is 70.15g/L in the fermented liquid, fermentating liquid volume is 3500L.
Comparing embodiment 4-2 (do not mend sugar, control condition is simplified, and first sugared concentration improves)
Use subtilis IFO 13621.With forming the 350L seed culture medium identical, that culture condition is identical with embodiment 4 substratum, cultivate after 18 hours for 36 ℃, access is by glucose 21.88% (w/v), dry yeast 1.0%, ammonium sulfate 0.7%, manganous sulfate 0.005%, lime carbonate 2.2% is in the 3200L fermention medium (pH 7.0) that each 50 μ g/ml of tryptophane, tyrosine and phenylalanine form.180 rev/mins of control mixing speed, 36 ℃ of culture temperature by 1: 0.8 ventilation, were cultivated 58 hours 36 ℃ of following aeration-agitations, and the content of D-ribose is 65.27g/L in the fermented liquid, and fermentating liquid volume is 3500L.
Claims (3)
1, a kind of regulate and control method of D-ribose fermenting process is characterized in that:
In 36～38 ℃ of scopes, tank pressure is at 0.5～0.8kg/cm with the culture temperature of FPC type distributed control system on-line Control D-ribose fermentation
2In the scope, mixing speed is in 180～220 rev/mins of scopes, the pH value scope of fermentation different steps: earlier fermentation pH6.50～7.0, fermentation middle and later periods pH6.00～6.50, the dissolved oxygen scope of fermentation different steps: earlier fermentation 18%～30%, fermentation middle and later periods 20%～35%, and timing sampling is measured Bacillus strain cell concn, concentration of substrate and product growing amount in the fermenting process, be worth 2.5% o'clock in sugared concentration subcritical, stream adds mends sugar; Wherein earlier fermentation is fermentation 0～18 hour, and the fermentation middle and later periods is for after fermenting 18 hours; Wherein stream adds and mends sugar and be glucose.
2, according to the process of claim 1 wherein that Bacillus strain in the said fermenting process is selected from one or more among bacillus pumilus CGMCC 0845, subtilis CGMCC 0846, bacillus pumilus ATCC 31093, the subtilis IFO 13621.
3, according to the method for claim 1 or 2, wherein said stream add the mode of mending sugar be with the 100-130 liter/hour flow velocity stream add glucose solution, make the glucose content in the fermention medium constant in the scope of 2.5%-3.0%, the concentration of the glucose solution that institute's stream adds is 30%-50%, and volume is the 10%-20% of fermentor tank volume.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02156496 CN1231592C (en) | 2002-12-18 | 2002-12-18 | D-ribose fermentation process regulating method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02156496 CN1231592C (en) | 2002-12-18 | 2002-12-18 | D-ribose fermentation process regulating method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1508256A CN1508256A (en) | 2004-06-30 |
| CN1231592C true CN1231592C (en) | 2005-12-14 |
Family
ID=34236239
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 02156496 Expired - Lifetime CN1231592C (en) | 2002-12-18 | 2002-12-18 | D-ribose fermentation process regulating method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1231592C (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104818346A (en) * | 2015-04-29 | 2015-08-05 | 宁夏诚志万胜生物工程有限公司 | Regulation and control method for fermentation process of L-arginine high in acid yield |
-
2002
- 2002-12-18 CN CN 02156496 patent/CN1231592C/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| CN1508256A (en) | 2004-06-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chao et al. | Bacterial cellulose production under oxygen-enriched air at different fructose concentrations in a 50-liter, internal-loop airlift reactor | |
| Ogbonna et al. | Sequential heterotrophic/autotrophic cultivation–an efficient method of producing Chlorella biomass for health food and animal feed | |
| Demirci et al. | Ethanol production by Saccharomyces cerevisiae in biofilm reactors | |
| Palamae et al. | Production of renewable biohydrogen by Rhodobacter sphaeroides S10: a comparison of photobioreactors | |
| JP2020534787A (en) | Continuous culture of black aspergillus and citric acid production method using it | |
| CN101709263B (en) | Chemostat continuous culture device | |
| Kondo et al. | Efficient production of acetic acid from glucose in a mixed culture of Zymomonas mobilis and Acetobacter sp. | |
| CN103224965A (en) | Method for producing pyrroloquinoline quinine through microbial fermentation and fermentation medium used in same | |
| CN102876743B (en) | Novel process for fermenting coenzyme Q10 based on online oxygen consumption rate control | |
| Du et al. | Production of L-lactic acid by Rhizopus oryzae in a bubble column fermenter | |
| CN101073394A (en) | Preparation of red leaven rice with high color value | |
| CN105543297B (en) | Hydrogenogen combines conversion of biomass and CO with alcaligenes eutrophus2The method for preparing polyhydroxyalkanoate | |
| CN117229982B (en) | Application of N- (1, 3-dimethylbutyl) -N' -phenyl p-phenylenediamine-quinone in culture of synechocystis | |
| Markov et al. | Spiral tubular bioreactors for hydrogen production by photosynthetic microorganisms: design and operation | |
| El-Diwany et al. | Effect of some fermentation parameters on ethanol production from beet molasses by Saccharomyces cerevisiae Y-7 | |
| CN1231592C (en) | D-ribose fermentation process regulating method | |
| CN1321174C (en) | Continuous flow culture method of industrial biological hydrogen preparing spawn and biological hydrogen preparing system reinforcing method | |
| CN109609557A (en) | A kind of wheat stalk based on fermentation process bacterium colony balance is stable, efficient producing gas method | |
| CN109536565A (en) | Method for producing succinic acid by utilizing mixed fermentation of high-temperature anaerobic bacteria for pyrolyzing sugar and actinobacillus succinogenes | |
| Karimi et al. | Mucor indicus as a biofilter and fermenting organism in continuous ethanol production from lignocellulosic hydrolyzate | |
| CN1027084C (en) | Process for preparing L-sorbose | |
| Zhang et al. | Isolation of xylose fermentation strains for ethanol production and xylose fermentation research | |
| CN108179112B (en) | Method for producing hydrogen by chlorella pyrenoidosa combined bacteria | |
| CN104611401A (en) | Method for improving microbial fermentation for production of 2, 3-butanediol | |
| Giridhar et al. | Repeated fed-batch sorbose fermentation by Gluconobacter oxydans |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: JIANGXI CHENGZHI BIOLOGICAL ENGINEERING Co.,Ltd. Assignor: Chengzhi Life Sci. & Tech. Co.,Ltd. Contract record no.: 2011360000083 Denomination of invention: D-ribose fermentation process regulating method Granted publication date: 20051214 License type: Exclusive License Open date: 20040630 Record date: 20110901 |
|
| CX01 | Expiry of patent term | ||
| CX01 | Expiry of patent term |
Granted publication date: 20051214 |