CN106701535B - A kind of Zhongshengmycin fermentation continuous feeding process units and its production method - Google Patents
A kind of Zhongshengmycin fermentation continuous feeding process units and its production method Download PDFInfo
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- CN106701535B CN106701535B CN201710037400.3A CN201710037400A CN106701535B CN 106701535 B CN106701535 B CN 106701535B CN 201710037400 A CN201710037400 A CN 201710037400A CN 106701535 B CN106701535 B CN 106701535B
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Abstract
The invention discloses a kind of Zhongshengmycin fermentation continuous feeding process units and its production methods, supplemented medium in Zhongshengmycin fermentation process is without the damp and hot sterilization of steam high temperature, transportation system, flowmeter metering, sterilization and filtration system acquisition asepsis feeding culture medium are prepared by supplemented medium, it continuously adds in fermentor, is normally carried out Zhongshengmycin fermentation lasts.Sterilization and filtration system obtains sterilization and pressure maintaining in fermentor while fermentation medium sterilization.The present invention dramatically reduces the labor intensity of employee, so that Zhongshengmycin fermentation process control of additive raw material is more flexible, accurate, has saved a large amount of steam, and it is horizontal to improve Zhongshengmycin fermenting and producing.There is not been reported for a kind of Zhongshengmycin fermentation continuous feeding process units of the present invention and its production method.
Description
Technical field
The present invention relates to a kind of fermenting and producing of biological pesticide, espespecially a kind of Zhongshengmycin fermentation continuous feeding process units
And its production method.
Background technique
According to the statistics of FAO (Food and Agriculture Organization of the United Nation) (FAO), during whole world agricultural and forestry production；Every year because disease pest and weed is made
At loss more than one third.3,000,000 chemical pesticide poisonings about occur every year for the whole world.In development China
Family, the whole world are annual because edible pollution by pesticides food causes collective poisoning event about 1650.Therefore, it greatly develops safe, low
It is malicious and nontoxic, low to accumulate and have become without accumulation, biological pesticide free from environmental pollution inevitable trend and whole world common recognition.Middle raw bacterium
Element is a kind of novel agricultural antibiotics biological pesticide, and application belongs to fungicide classification, for preventing and treating corps diseases.It
Producing strains be it is isolated from the soil in China Hainan Island, be named as Strepiomyces lavendulae Hainan mutation (Strepromyces
Lavenduae hainanensis n.var).As a kind of agricultural anti-microbial type biological pesticide to gram-positive bacteria, gram
Negative bacterium and filamentous fungi have strong inhibition or killing effect.To bacterial blight of rice, soft rot of cabbage, ring rot of apple
There is good preventive effect with apple leaf spot etc..But it is safety low-poison to human and livestock, it is compatible with the environment friendly.Zhongshengmycin (is changed
Scientific name: 1-N glucoside base-2-amino of streptolidine base L-- 2 deoxidation gulose amine of lysine；Molecular formula: C19H34O8N8；Molecule
Amount: 502.0) exactly having the characteristics that high-efficiency low-toxicity, low accumulation, noresidue, by the peptide bond for inhibiting pathogenetic bacteria protein
It generates, eventually leads to bacterial death；The growth of mycelia can inhibit to fungi, inhibit the sprouting of spore, play prevention and treatment fungoid disease
Harmful effect；It is the indispensable medication of nuisanceless fruit tree, vegetables production base.It is widely used in preventing and treating bacterial blight of rice, apple
Ring spot, soft rot of cabbage etc..Therefore, a kind of Zhongshengmycin fermentation continuous feeding process units and production method, reduction are developed
Energy consumption improves Zhongshengmycin fermenting and producing level, and then improves Zhongshengmycin yield with good economic and social profit.
Zhongshengmycin is soybean cake powder, cornstarch, corn flour, Portugal using the mutation of Strepiomyces lavendulae Hainan as producing strains
Grape sugar, calcium carbonate, sodium chloride, ammonium chloride, magnesium sulfate, glutamic acid, arginine, piece alkali etc. are that primary raw material is obtained by fermented and cultured
The secondary metabolite arrived.The main method of Zhongshengmycin fermentation at present are as follows: the culture of Zhongshengmycin producing strains shake-flask seed →
It is inoculated into seeding tank expansion and cultivates → be inoculated into ferment tank culture generation Zhongshengmycin, in 50 hours ~ 240 hours hairs
One to five fed-batch is carried out in the ferment period.
At present in place of technique Shortcomings:
1, current supplemented medium use moist heat sterilization method known in the industry, due to moist heat sterilization to reach 121 DEG C,
Sterilizing in 30 minutes is held time, and the pyroprocess for causing supplemented medium to be in 80 DEG C or more will be when 60 minutes or more
Between, it causing supplemented medium appearance after long-time high temperature rubescent or brown is presented, the nutriment of supplemented medium is destroyed,
The substance toxic to Zhongshengmycin producing strains is even generated, the raising of Zhongshengmycin fermentation level is influenced.
2, current supplying technics are a large amount of feed supplements of interval, cause nutrient media components fluctuation in fermentor larger, to middle life
Micro- growing environment of rhzomorph producing strains causes strong influence, is unfavorable for the growth of producing strains and the growth of secondary metabolite.
3, a large amount of feed supplements of current fermentation process interval, easily form glucose effect known in the industry, cause to secondary generation
It thanks to the inhibition of product generation and influences the growth of Zhongshengmycin yield.
4, a large amount of feed supplements of current interval make the prometaphase overnutrition that ferments, mass propagation thallus；Middle and later periods nutrition is not
Foot, can not further increase Zhongshengmycin fermentation level, Zhongshengmycin content is hovered in 8.00g/L -10.00g/L in fermentation liquid
Between.
5, a large amount of supplying technics of current interval need to consume a large amount of steam carry out multiple supplemented medium disinfection and
The disinfection of multiple feed supplement pipeline, staff labor intensity is big and mass energy is caused to consume.
Summary of the invention
Technical problem to be solved by the present invention lies in provide a kind of Zhongshengmycin fermentation continuous feeding device and its production
It is horizontal that Zhongshengmycin fermenting and producing can be improved by the continuous feeding device and its production method in method；Mitigate the labor of employee
Fatigue resistance；It is energy-saving that the realization of steam usage amount can also be reduced.
To solve the above problems, the technical solution of the invention is as follows:
1, a kind of Zhongshengmycin fermentation continuous feeding process units, including sterilization and filtration system, flowmeter and feed-batch culture
Basigamy transportation system.
(1), the outlet of the sterilization and filtration system of the continuous feeding device passes through pipeline and fermentor feed supplement conduit entrance
The import of connection, sterilization and filtration system is connect by pipeline with flowmeter outlet, and flowmeter import passes through pipeline and feed-batch culture
The outlet of basigamy transportation system connects.
(2), the liquid fine filter and a precision that the sterilization and filtration system is 0.22 μ by 2 filtering accuracies are 0.45 μ
Liquid prefilter formed by placed in series, liquid prefilter be sterilization and filtration system input end, sterilization and filtration system into
Mouth pipeline connects the first valve and the second valve by the first threeway and pipeline.
(3), it is connected by the first valve through pipeline between the sterilization and filtration system import and flowmeter outlet.
(4), the flowmeter import connects the second threeway outlet through pipeline.
(6), the import of the second threeway of the connection flowmeter import connects supplemented medium by pipeline and prepares conveying
The outlet of the dnockout pumps of system.
(7), the import of the dnockout pumps connects the outlet of third threeway by pipeline.
(8), an import of the third threeway of the connection dnockout pump inlet connects the outlet of third valve by pipeline,
The import of the third valve is connect by pipeline with the first supplemented medium ingredient pot bottom.
(9), another import of the third threeway of the connection dnockout pump inlet connects going out for the 4th valve by pipeline
Mouthful, the import of the 4th valve is connect by pipeline with the second supplemented medium ingredient pot bottom.
(10), another outlet of the second threeway of the connection flowmeter import connects the 4th threeway by pipeline
Import；Two outlets of the 4th threeway respectively connect the 5th valve and the 6th valve, the 5th valve and the 6th by pipeline
Valve all respectively connects a pipeline correspondence and returns in respective supplemented medium material-compound tank.
2, a kind of production method of Zhongshengmycin fermentation continuous feeding, including following technical process:
(1), the sterilizing of sterilization and filtration system and associated connecting pipe: the side of real tank sterilization known in the industry is used
Method is in fermentor while culture medium sterilization, to the liquid of the liquid fine filter of 2 0.22 μ of precision, 1 0.45 μ of precision
Body prefilter and associated connecting pipe carry out live steam sterilization.
(2), the pressure maintaining after sterilization and filtration system and associated connecting pipe sterilize: sterilizing terminates to utilize the nothing in fermentor
Liquid prefilter and associated communicating pipe of the bacterium air to the liquid fine filter of 2 concatenated 0.22 μ of precision, 1 0.45 μ of precision
Positive pressure after road is sterilized is kept.
(3), the preparation of supplemented medium: by a certain proportion of glucose, sodium chloride, ammonium chloride, magnesium sulfate, biphosphate
Potassium, glutamic acid, it is molten match after arginine in two supplemented medium material-compound tanks plus drinking water stirring, be completely dissolved, be configured to
Certain concentration.
(4), feed supplement: certain density supplemented medium in the first supplemented medium material-compound tank is exported with dnockout pumps, warp
Overcurrent meter, 1 0.45 μ liquid prefilter of precision, 2 0.22 μ of precision liquid fine filter sterilizing after, reach sterile feed supplement
Culture medium, which is added, sets out in fermentation tank.
(5), supplemented medium flows back: for the flow for controlling supplemented medium, the supplemented medium part of dnockout pumps output
It is back in the first supplemented medium material-compound tank through return line flow.
(6), continuous feeding: 2 supplemented medium preparing tanks replace ingredients, feed supplement, by feeding-system third valve and
The switching control of 4th valve is realized to a small amount of multiple fed-batch of fermentor or continuous flow feeding.
The liquid fine filter and associated communicating pipe that 1 precision is 0.45 μ liquid prefilter, 2 precision are 0.22 μ
The sterilizing in road carries out all to synchronize when the sterilizing of every batch of fermentation tank culture medium.
The 0.45 μ liquid prefilter filter core material of precision, 0.22 μ liquid fine filter filter core material of precision are all tolerable to be made
Reached tens of times with 121 DEG C of steam realization, sterilizing in 30 minutes.
The material of the 0.22 μ liquid fine filter filter core of precision be PES(polyether sulfone), NL(nylon 6), PTFE(polytetrafluoro
Ethylene), PVDF(polyvinylidene fluoride).
The material of the 0.45 μ liquid prefilter filter core of precision be PES(polyether sulfone), NL(nylon 6), PTFE(polytetrafluoro
Ethylene), PVDF(polyvinylidene fluoride), PP(polypropylene).
The supplemented medium be formulated as by feed supplement glucose, sodium chloride, ammonium chloride, magnesium sulfate, potassium dihydrogen phosphate,
Glutamic acid, it is molten match after arginine investment fill in the supplemented medium material-compound tank of a certain amount of drinking water, stirring makes feed-batch culture
Base sufficiently dissolves, and adds drinking water, and the supplemented medium in supplemented medium material-compound tank is settled to respectively containing glucose
0.00% -35.00%, sodium chloride-containing 0.00% -5.00%, containing ammonium chloride 0.00% -6.00%, containing magnesium sulfate 0.00% -6.50%,
Phosphoric acid potassium dihydrogen 0.00% -3.50% contains glutamic acid 0.00% -6.50%, contains arginine 0.00% -7.50%.
The continuous feeding be fermentor in concentration of reduced sugar≤2.50% when, will be in the first supplemented medium material-compound tank
Prepared supplemented medium passes through flowmeter, 1 0.45 μ liquid prefilter of precision, 2 precision by dnockout pumps output
0.22 μ liquid fine filter sterilizing filtering enter fermentor in, maintain fermentor in concentration of reduced sugar 0.50% -2.50% it
Between.
Supplemented medium reflux enter in fermentor to guarantee the flow of supplemented medium as required and by dnockout
Pump output supplemented medium more than needed is returned in former supplemented medium material-compound tank by by-pass line.
The continuous feeding is to guarantee that the concentration of reduced sugar in fermentor maintains a concentration ranges, can be lacked at any time
Measure that multiple intermittent injecting more continuously continuously adds drinking water or the supplemented medium of >=a kind of different ratio enters fermentation
The process of tank, is used alternatingly using 2 supplemented medium material-compound tanks, and feed supplement type, feed supplement speed are flexibly grasped according to process requirements
Degree and feed supplement time, feed supplement time can put the overall process of tank from fermentation incipient " 0 " hour to fermentation ends.
After adopting the above scheme, the present invention has following distinguishing feature compared with prior art technology:
1, the supplemented medium of Zhongshengmycin continuous feeding of the invention does not need the moist heat sterilization of steam, reduces feed supplement
Destruction and formation caused by culturing gene high-temperature heat sterilization are to the harmful substance of Zhongshengmycin producing strains.
2, the continuous feeding can maintain the relatively stable of fermentation liquid microenvironment in Zhongshengmycin fermentation process, avoid because
The a large amount of feed supplements of interval cause the variation of fermentation liquid microenvironment, influence or change Zhongshengmycin producing strains eubolism approach.
3, the continuous feeding can control the concentration of fermentation liquid glucose, avoid the glucose of high concentration to Zhongshengmycin
Producing strains generate Zhongshengmycin and unfavorable inhibition are caused to influence, and are conducive to extend the time of Zhongshengmycin producing strains cometabolism,
Improve Zhongshengmycin fermentation level.
4, the continuous feeding can flexibly grasp Zhongshengmycin fermentation feed medium type, feed supplement according to process requirements
Speed and feed supplement time, realize being precisely controlled for fermentation process.
5, Zhongshengmycin continuous feeding culture medium of the invention does not need the moist heat sterilization of steam, has saved a large amount of feed supplement
The steam for the multiple sterilization of feed supplement pipeline that culture medium sterilization and a large amount of feed supplements of interval need, while reducing employee's
Labor intensity has saved manpower and material resources, has good economic and social profit.
Detailed description of the invention
Fig. 1 is Zhongshengmycin fermentation continuous feeding device and its production method schematic diagram of the present invention:
Specific embodiment
The present invention is described in further detail in the following with reference to the drawings and specific embodiments.
It ferments continuous feeding process units disclosed is a kind of Zhongshengmycin, as shown in Figure 1, for institute of the present invention
State Zhongshengmycin fermentation continuous feeding process units schematic diagram.The process units of the Zhongshengmycin continuous feeding includes: hair
Fermentation tank 1, liquid fine filter 2, liquid fine filter 3, liquid prefilter 4, the second valve 5, the first valve 6, flowmeter 7, the 5th valve
Door the 8, the 6th valve 9, the first supplemented medium material-compound tank 10, the second supplemented medium material-compound tank 11, third valve 12, the 4th valve
Door 13, dnockout pumps 14.Wherein: the import of the fermentor 1 is connect with the outlet of liquid fine filter 2 by pipeline, the liquid refined filtration
The import of device 2 is connect with the outlet of liquid fine filter 3 by pipeline, the import of the liquid fine filter 3 and going out for liquid prefilter 4
Mouth is connected by pipeline, and the import of the liquid prefilter 4 and the import of the second valve 5 and the outlet of the first valve 6 pass through pipeline
Connection, the outlet of second valve 5 is connected by pipeline to be emptied, and the import of first valve 6 and the outlet of flowmeter 7 pass through pipe
Road connection, the import of the flowmeter 7 and the outlet of dnockout pumps 14 and the import of the 5th valve 8 and the 6th valve 9 pass through pipeline
The outlet of connection, the 5th valve 8 is entered in the first supplemented medium material-compound tank 10 by pipeline, the outlet of the 6th valve 9
Entered in the second supplemented medium material-compound tank 11 by pipeline, the outlet of the first supplemented medium material-compound tank 10 and third valve
12 import is connected by pipeline, and the outlet of the second supplemented medium material-compound tank 11 and the import of the 4th valve 13 pass through pipeline
Connection, the outlet of the third valve 12 and the outlet of the 4th valve 13 are connect with the import of dnockout pumps 14 by pipeline.
The present invention also discloses a kind of Zhongshengmycin fermentation continuous feeding production method, as shown in Figure 1, the production method work
Skill process are as follows:
(1) sterilization of liquid fine filter 2, liquid fine filter 3, liquid prefilter 4: with moise-heat sterilization known in the industry
The method of sterilizing carries out sterilize in batches during culture medium in fermentor, closes the first valve 6, the second valve of pico- unlatching
5, guarantee that liquid fine filter 2, liquid fine filter 3, liquid prefilter 4, the second valve 5 have live steam to be full of and control flow row
Out, reach 121 DEG C, the temperature of 30 minutes sterilizations with hold time.
(2) the sterile pressure maintaining of liquid fine filter 2, liquid fine filter 3, liquid prefilter 4: medium sterilization knot in fermentor
Tank body pressure >=0.02Mpa is maintained with filtrated air after beam, simultaneously closes off the second valve 5, and maintain in entire fermentation process
Ferment pressure tank >=0.02Mpa, maintains pressure >=0.02Mpa of liquid fine filter 2, liquid fine filter 3, liquid prefilter 4.
(3) supplemented medium prepares one: closing third valve 12, squeezes into one in advance in the first supplemented medium material-compound tank 10
Quantitative drinking water puts into supplemented medium load weighted in proportion in advance, opens stirring, add drinking water, it is ensured that first mends
Supplemented medium in material culture medium material-compound tank 10 is completely dissolved and reaches required volume and matched proportion density.
(4) supplemented medium prepares two: closing the 4th valve 13, squeezes into one in advance in the second supplemented medium material-compound tank 11
Quantitative drinking water puts into supplemented medium load weighted in proportion in advance, opens stirring, add drinking water, it is ensured that second mends
Supplemented medium in material culture medium material-compound tank 11 is completely dissolved and reaches required volume and matched proportion density.
(5) feed supplement one: the numerical value of fermentation liquid reduced sugar in fermentor is measured by sampling, when the reduced sugar of fermentation liquid in fermentor
When reaching the range for needing feed supplement, third valve 12 is opened, the 5th valve 8 is opened, closes the 6th valve 9, close the 4th valve
13 unlatching dnockout pumps 14 make the supplemented medium in the first supplemented medium material-compound tank 10 pass through third valve 12, dnockout pumps
14, the 5th valve 8 is recycled；The open degree slowly opened the first valve 6, adjust the 5th valve 8, so that the stream of flowmeter 7
Amount reaches supplying technics requirement, and culture medium concentration of reduced sugar is 0.50% ~ 2.50% in maintenance fermentor.
(6) feed supplement two: when the supplemented medium in the first supplemented medium material-compound tank 10 be finished or technique require add it
When his supplemented medium, switching uses the supplemented medium in the second supplemented medium material-compound tank 11；The 4th valve 13 is opened, is closed
Third valve 12 is closed, the 6th valve 9 is opened, closes the 5th valve 8, dnockout pumps 14 is maintained to open, adjusts the first valve 6 and the 6th
The open degree of valve 9 maintains culture medium reduced sugar in fermentor dense so that the flow of flowmeter 7 reaches the requirement of supplying technics
Degree is 0.50% ~ 2.50%.
(7) continuous feeding: according to the supplying technics needs of fermentation process, according to step (3), step (4) in the first feed supplement
Culture medium material-compound tank 10, the second supplemented medium material-compound tank 11 carry out alternately preparing various supplemented mediums；According to step (5),
Supplemented medium in step (6) switching the first supplemented medium material-compound tank 10 of conveying or the second supplemented medium material-compound tank 11 connects
It is continuous to add the needs that Zhongshengmycin fermenting and producing is maintained in fermentor.
(8) fermentation ends clean 2 supplemented medium material-compound tanks, dnockout pumps, flowmeter, liquid pre-flock using drinking water
Device, 2 liquid fine filters and associated pipeline valve, it is ensured that the cleaning of continuous feeding device is stand-by.
Illustrate the production method of Zhongshengmycin fermentation continuous feeding of the present invention below by specific embodiment
Embodiment 1
Zhongshengmycin fermented and cultured, the continuous feeding device of fermentor culture medium in fermentor are carried out with 30L fermentor
Carry out disinfection sterilizing while sterilization, and the continuous feeding device of fermentor is in sterile during Zhongshengmycin fermented and cultured
State of pressure keeping.For Zhongshengmycin fermented and cultured to 15 hours, measuring the content of reducing sugar in fermentation liquid was 2.32%, starts to add and contains
Glucose 15.00%, sodium chloride-containing 0.50%, the benefit of containing ammonium chloride 0.55%, containing magnesium sulfate 0.05%, phosphoric acid potassium dihydrogen 0.01%
Expect culture medium.Controlling feed-batch culture Dryweather flow is≤2.50L/h, every 1 hour measurement Zhongshengmycin fermentation liquid content of reducing sugar,
Content of reducing sugar stopping feed supplement in 10 hours before 1.50% ~ 2.00%, continuous feeding to fermentation ends in the fermentation liquid is controlled, is fermented
Liquid continues fermented and cultured to fermentation ends, and fermentation ends put tank, and it is 0.85% that tank fermentation liquid content of reducing sugar is put in measurement, puts tank hair
Zymotic fluid Zhongshengmycin content 11.33g/L.
Embodiment 2
Zhongshengmycin fermented and cultured, the continuous feeding device of fermentor culture medium in fermentor are carried out with 30L fermentor
Carry out disinfection sterilizing while sterilization, and the continuous feeding device of fermentor is in sterile during Zhongshengmycin fermented and cultured
State of pressure keeping.For Zhongshengmycin fermented and cultured to 25 hours, measuring the content of reducing sugar in fermentation liquid was 1.96%, starts to add and contains
Glucose 20.00%, containing ammonium chloride 0.65%, containing magnesium sulfate 0.04%, phosphoric acid potassium dihydrogen 0.01%, contains paddy at sodium chloride-containing 0.55%
Propylhomoserin 0.03%, the supplemented medium containing arginine 0.02%.Controlling feed-batch culture Dryweather flow is≤2.50L/h, is surveyed every 1 hour
Determine Zhongshengmycin fermentation liquid content of reducing sugar, controls content of reducing sugar in Zhongshengmycin fermentation liquid and continuously mended 1.30% ~ 1.80%
Material is to stopping feed supplement in 5 hours before fermentation ends, and fermentation liquid continues fermented and cultured to fermentation ends, and fermentation ends put tank, and measurement is put
Tank fermentation liquid content of reducing sugar is 1.05%, puts tank fermentation liquid Zhongshengmycin content 10.56g/L.
Embodiment 3
Zhongshengmycin fermented and cultured, the continuous feeding device of fermentor culture medium in fermentor are carried out with 30L fermentor
Carry out disinfection sterilizing while sterilization, and the continuous feeding device of fermentor is in sterile during Zhongshengmycin fermented and cultured
State of pressure keeping.For Zhongshengmycin fermented and cultured to 18 hours, measuring the content of reducing sugar in fermentation liquid was 2.15%, starts to add and contains
Glucose 35.00%, sodium chloride-containing 0.45%, containing ammonium chloride 0.55%, containing magnesium sulfate 0.03%, phosphoric acid potassium dihydrogen 0.02%, paddy ammonia
Sour 0.05%, the supplemented medium containing arginine 0.04%.Controlling feed supplement flow is≤3.00L/h, every raw bacterium in measurement in 1 hour
Plain fermentation liquid content of reducing sugar, controlling content of reducing sugar in Zhongshengmycin fermentation liquid, 2.00% ~ 2.50%, continuous feeding 30 is small
When；Continue fermentation and be down to 1.30% ~ 1.50% to Zhongshengmycin fermentation liquid reduced sugar, starts to add the feed supplement containing glucose 15.00%
Culture medium, control feed-batch culture Dryweather flow are≤3.00L/h, every 1 hour measurement Zhongshengmycin fermentation liquid content of reducing sugar, control
Zhongshengmycin fermentation liquid content of reducing sugar processed is between 1.20% ~ 1.60%, and stopping in 8 hours is mended before continuous feeding to fermentation ends
Material, fermentation liquid continue fermented and cultured to fermentation ends, and fermentation ends put tank, and it is 0.72% that tank fermentation liquid content of reducing sugar is put in measurement,
Put tank fermentation liquid Zhongshengmycin content 12.21g/L.
Embodiment 4
Zhongshengmycin fermented and cultured, the continuous feeding device of fermentor culture medium in fermentor are carried out with 30L fermentor
Carry out disinfection sterilizing while sterilization, and the continuous feeding device of fermentor is in sterile during Zhongshengmycin fermented and cultured
State of pressure keeping.For Zhongshengmycin fermented and cultured to 10 hours, measuring the content of reducing sugar in fermentation liquid was 3.16%, started to add not
Supplemented medium containing any other raw and auxiliary material containing only drinking water, control feed-batch culture Dryweather flow is≤3.50L/h, every 1
Hour measurement Zhongshengmycin fermentation liquid content of reducing sugar, controls in Zhongshengmycin fermentation liquid content of reducing sugar 2.00% ~ 2.50%,
Start to add containing glucose 25.00%, sodium chloride-containing 0.55%, containing ammonium chloride 0.55%, containing magnesium sulfate 0.04%, phosphoric acid potassium dihydrogen
0.02%, the supplemented medium containing arginine 1.15%.Control feed-batch culture Dryweather flow be≤3.50L/h, continuous feeding 30 hours,
Continue fermentation and be down to 1.10% ~ 1.30% to Zhongshengmycin fermentation liquid reduced sugar, starts to add the feed supplement training containing glucose 10.00%
Base is supported, control feed-batch culture Dryweather flow is≤3.50L/h, every 1 hour measurement Zhongshengmycin fermentation liquid content of reducing sugar, control
Fermentation liquid content of reducing sugar is between 1.00% ~ 1.30%, stopping feed supplement in 6 hours before continuous feeding to fermentation ends, fermentation liquid after
Continue fermented and cultured to fermentation ends, fermentation ends put tank, and it is 0.81% that tank fermentation liquid content of reducing sugar is put in measurement, put tank fermentation liquid
Zhongshengmycin content 12.56g/L.
Embodiment 5
Zhongshengmycin fermented and cultured, the continuous feeding device of fermentor culture medium in fermentor are carried out with 30L fermentor
Carry out disinfection sterilizing while sterilization, and the continuous feeding device of fermentor is in sterile during Zhongshengmycin fermented and cultured
State of pressure keeping.For Zhongshengmycin fermented and cultured to 12 hours, measuring the content of reducing sugar in fermentation liquid was 2.88%, started to add not
Supplemented medium containing any raw and auxiliary material containing only drinking water.Controlling feed-batch culture Dryweather flow was≤3.00L/h, every 1 hour
Zhongshengmycin fermentation liquid content of reducing sugar is measured, content of reducing sugar in Zhongshengmycin fermentation liquid is controlled and starts 2.00% ~ 2.50%
It adds containing glucose 30.00%, phosphoric acid potassium dihydrogen 0.02%, glutamic acid 0.86%, the supplemented medium containing arginine 1.15%.Control
Feed-batch culture Dryweather flow processed is≤3.00L/h, continuous feeding 35 hours, continues fermentation to Zhongshengmycin fermentation liquid reduced sugar and is down to
1.30% ~ 1.50%, start to add the supplemented medium containing glucose 15.00%, control feed-batch culture Dryweather flow is≤3.00L/h,
Every 1 hour measurement Zhongshengmycin fermentation liquid content of reducing sugar, fermentation liquid content of reducing sugar was controlled between 0.80% ~ 1.30%,
Stopping feed supplement in 5 hours before continuous feeding to fermentation ends, fermentation liquid continue fermented and cultured to fermentation ends, and tank fermentation liquid is put in measurement
Content of reducing sugar is 0.88%, puts tank fermentation liquid Zhongshengmycin content 11.88g/L.
The measuring method of the fermentation liquid content of reducing sugar uses fermentation liquid content of reducing sugar measuring method known in the industry.
The measuring method of the fermentation liquid Zhongshengmycin content are as follows:
(1) bioassay: antibiotic-microbial assay.
(2) test organisms of bacteria suspension: bacillus subtilis [CMCC(B) 63501].
The supplemented medium material-compound tank of described schematic diagram Fig. 1 can be 2 and 2 or more, dense for different formulations and difference
The supplemented medium of degree is prepared, so that flexibly the supplemented medium of various needs is added in control in fermentation process.
The continuous feeding device of described schematic diagram Fig. 1 can be 1 set and 1 set or more and connect with same fermentor, realize each
Plant the function of single or compound supplemented medium flexible feed supplement simultaneously.
The continuous feeding device of described schematic diagram Fig. 1 can be expanded into before flowmeter, be connected to forage feeding station with pipeline, be led to
Cross function of the control realization to 1 and 1 or more the flexible continuous feeding of fermentor of forage feeding station valve.
The foregoing is merely general examples of the invention, are not used to limit the scope of implementation of the present invention.Therefore in every case
The changes or modifications that claim under this invention and specification are done all should belong within the scope of the invention patent covers.
Claims (9)
- The continuous feeding process units 1. a kind of Zhongshengmycin ferments, is primarily characterized in that: including fermentor, aseptic filtration system System, flowmeter and supplemented medium prepare transportation system；The liquid that the liquid fine filter and a precision that the sterilization and filtration system is 0.22 μ by 2 filtering accuracies are 0.45 μ is pre- Filter is formed by placed in series, liquid prefilter be sterilization and filtration system input end, sterilization and filtration system inlet pipeline by First threeway and pipeline connect the first valve and the second valve；It is mended by pipeline and fermentor the outlet of the sterilization and filtration system Pipe material import connection, is connected by wherein the first valve through pipeline between the sterilization and filtration system import and flowmeter outlet； The flowmeter import connects the second threeway outlet through pipeline；The supplemented medium is prepared transportation system and is made of two supplemented medium material-compound tanks and a dnockout pumps, two feed supplements Culture medium material-compound tank is respectively the first supplemented medium material-compound tank, the second supplemented medium material-compound tank, the wherein import of dnockout pumps The outlet of third threeway is connected by pipeline；One import of the third threeway of the connection dnockout pump inlet is connected by pipeline The import of the outlet of one third valve, the third valve is connect by pipeline with the first supplemented medium ingredient pot bottom；Institute Another import for stating the third threeway of connection dnockout pump inlet connects the outlet of the 4th valve by pipeline, the 4th valve Import is connect by pipeline with the second supplemented medium ingredient pot bottom；Another outlet of second threeway of the connection flowmeter import connects the import of the 4th threeway by pipeline；4th Two outlets of threeway respectively connect the 5th valve and the 6th valve by pipeline, and the 5th valve connects one article of pipeline and returns to first Supplemented medium material-compound tank is interior, the 6th valve connects one article of pipeline and returns in the second supplemented medium material-compound tank；4th threeway Import by pipeline connect supplemented medium prepare transportation system dnockout pumps outlet.
- The continuous feeding process units 2. a kind of Zhongshengmycin according to claim 1 ferments, is primarily characterized in that: feed supplement Culture medium material-compound tank is 2 or more, can alternately switch ingredient feed supplement, realize continuous feeding.
- The continuous feeding process units 3. a kind of Zhongshengmycin according to claim 1 ferments, is primarily characterized in that: described 0.45 μ liquid prefilter filter core material of precision, 0.22 μ liquid fine filter filter core material of precision are all tolerable to be realized using steam It sterilizes for 121 DEG C, 30 minutes and reaches tens of times.
- The continuous feeding process units 4. a kind of Zhongshengmycin according to claim 1 ferments, is primarily characterized in that: described The material of 0.22 μ liquid fine filter filter core of precision is PES(polyether sulfone), NL(nylon 6), PTFE(polytetrafluoroethylene (PTFE)), PVDF(it is poly- Vinylidene fluoride).
- The continuous feeding process units 5. a kind of Zhongshengmycin according to claim 1 ferments, is primarily characterized in that: described The material of 0.45 μ liquid prefilter filter core of precision is PES(polyether sulfone), NL(nylon 6), PTFE(polytetrafluoroethylene (PTFE)), PVDF(it is poly- Vinylidene fluoride), PP(polypropylene).
- 6. a kind of production method of Zhongshengmycin fermentation continuous feeding process units according to claim 1, mainly walks Suddenly it is:(1), the sterilizing of fermentor, sterilization and filtration system and associated connecting pipe: with moise-heat sterilization known in the industry sterilizing Method carries out sterilize in batches during culture medium in fermentor, closes between sterilization and filtration system import and flowmeter outlet The first valve, pico- the first threeway connection for opening sterilization and filtration system inlet pipeline the second valve, guarantee 2 precision Second valve of the liquid fine filter of 0.22 μ, the liquid prefilter of 1 0.45 μ of precision and the first threeway connection, there is live steam Be full of and control flow discharge, reach 121 DEG C, the temperature of 30 minutes sterilizations with hold time；(2), the pressure maintaining after sterilization and filtration system and the sterilizing of associated connecting pipe: after step (1) sterilizes, the is simultaneously closed off Second valve of one threeway connection, using the filtrated air in fermentor to 2 concatenated liquid fine filters, 1 liquid pre-flock Positive pressure after device and associated connecting pipe are sterilized is kept；(3), the preparation of supplemented medium: by a certain proportion of glucose, sodium chloride, ammonium chloride, magnesium sulfate, potassium dihydrogen phosphate, Glutamic acid, it is molten match after arginine in two supplemented medium material-compound tanks plus drinking water stirring, be completely dissolved, be configured to it is certain Concentration；(4), feed supplement: will wherein certain density supplemented medium is exported with dnockout pumps in the first supplemented medium material-compound tank, pass through Overcurrent meter, 1 0.45 μ liquid prefilter of precision, 2 0.22 μ of precision liquid fine filter sterilizing after, reach sterile feed supplement Culture medium, which is added, sets out in fermentation tank；(5), supplemented medium flows back: for the flow for controlling supplemented medium, the supplemented medium of dnockout pumps output is according to flow The requirement of meter, maintain fermentor in culture medium concentration of reduced sugar within a preset range, part supplemented medium more than needed is through second Threeway is flow back into the first supplemented medium material-compound tank along the 4th threeway；(6), continuous feeding: the ingredients of 2 supplemented medium preparing tank alternate steps (3), step (4), (5) feed supplement and return Stream, realized by the switching control that supplemented medium prepares transportation system valve to a small amount of multiple fed-batch of fermentor or Continuous flow feeding.
- The continuous feeding production method 7. a kind of Zhongshengmycin according to claim 6 ferments, is primarily characterized in that: can be with When add the supplemented medium of drinking water and >=a kind of different ratio, supplemented medium material-compound tank is used alternatingly, and flexibly grasps feed supplement Type, feed rate and feed supplement time, feed supplement time put the overall process of tank from fermentation incipient " 0 " hour to fermentation ends.
- The continuous feeding production method 8. a kind of Zhongshengmycin according to claim 6 ferments, is primarily characterized in that: step (5) in, when the reduced sugar of culture medium in fermentor reaches the range for needing feed supplement, the first supplemented medium material-compound tank is opened Third, the 5th valve close the four, the 6th valves of the second supplemented medium material-compound tank, open dnockout pumps and make the first feed supplement Supplemented medium in culture medium material-compound tank is recycled by third, the 5th valve, dnockout pumps；Slowly open aseptic filtration system The unlatching of the bottom third valve of the first valve, the first supplemented medium material-compound tank of adjustment between system import and flowmeter outlet Degree so that the flow of flowmeter reaches supplying technics requirement, maintain in fermentor culture medium concentration of reduced sugar 0.50% ~ 2.50%。
- The continuous feeding production method 9. a kind of Zhongshengmycin according to claim 6 ferments, is primarily characterized in that: step (6) in, when the supplemented medium in the first supplemented medium material-compound tank is finished or technique requires to add other supplemented mediums, Switching uses the supplemented medium in the second supplemented medium material-compound tank；The of the second supplemented medium material-compound tank is opened at this time Four, the 6th valve closes third, the 5th valve of the first supplemented medium material-compound tank, dnockout pumps is maintained to open, adjusts degerming The 4th valve of bottom of the first valve, the second supplemented medium material-compound tank of adjustment between filter system inlets and flowmeter outlet Open degree, so that the flow of flowmeter reaches the requirement of supplying technics, culture medium concentration of reduced sugar is 0.50% in maintenance fermentor ~2.50%。
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| CN1772877A (en) * | 2005-09-28 | 2006-05-17 | 哈尔滨工业大学 | Continuous flow culture method of industrial biological hydrogen preparing spawn and biological hydrogen preparing system reinforcing method |
| CN103566384A (en) * | 2013-10-10 | 2014-02-12 | 宁夏启元药业有限公司 | Deactivating method of vitamin C one-step fermentation broth |
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