CN101619302A - Liquid fermentation process of bacillus megatherium - Google Patents
Liquid fermentation process of bacillus megatherium Download PDFInfo
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Abstract
The invention discloses a liquid fermentation process of bacillus megatherium, comprising the following steps: (1) proportioning: the concentration of a solid of a supplementary material is 30-35 percent, and the pH value is 6.2-7.0; (2) sterilizing a seeding tank: the temperature is 115-126 DEG C, the pressure of the tank is 0.11-0.13MPa, and the time is 30-40 min; (3) inoculating seeds according to 1-2 percent of the mass ratio of the supplementary material at the temperature of 26-30 DEG C; (4) fermenting the seeding tank: the temperature is 28-30 DEG C, the fluxes of sterile air before and after 10h are respectively 0.4-0.6 and 0.9-1.1 supplementary material volume/h, and the temperature is 40-48 DEG C; (5) sterilizing a fermentation tank: the process is same as the step (2); (6) inoculating seeds in the seeding tank according to 8-12 percent of the mass ratio of the supplementary material, other processes are same as the step (3); (7) fermenting the fermentation tank: the process is same as the step (4); and (8) discharging. The whole process flow has high efficiency, less pollution, low reladling rate and reasonable flow and can be widely used for producing bacillus megatherium liquid and solid products (needing to combine a dry process) in batch.
Description
Technical field
The present invention relates to a kind of liquid fermentation process, more specifically to the liquid fermentation process of a kind of bacillus megaterium.
Background technology
Existing bacillus megaterium breeds in soil.This bacterial classification in process of growth, can dissociate in a large number have cured in the soil can not be improved the utilization ratio of chemical fertilizer by the nitrogen of crop absorption, phosphorus, potassium in a large number, effectively alleviate because the negative effects such as soil compaction that a large amount of use chemical fertilizer cause.Simultaneously, can generate multiple growth parahormone again, promote plant growth, improve its degeneration-resistant proterties.Bacillus megaterium can also produce multiple antibiotic, suppresses the germ growth, and it is highly beneficial that this uploads germ to elimination.
But, in the soil bacillus megaterium quantity of self-sow very little, insignificant.And the domestic artificial production that does not also have this type of bacterial classification does not at present just have corresponding production technique yet.
Summary of the invention
Technical problem to be solved by this invention provides a kind of liquid fermentation process efficiently scale operation, sophisticated bacillus megaterium.
In order to solve the problems of the technologies described above, the liquid fermentation process of bacillus megaterium provided by the invention comprises the steps:
(1) batching: protein and carbohydrate supplementary material are made up of by the starch of 10-12%, the sucrose of 4-6%, the peptone of 1-3%, the yeast extract paste of 0.1-0.3, the ammonium sulfate of 0.8-1.2%, sal epsom and remaining water of 0.4-0.6% mass ratio, with protein and carbohydrate supplementary material, water and sodium hydroxide feed material mixing tank, mix while sending, until mixing, the solids concentration that makes supplementary material is 30-35%, and the pH value of supplementary material is 6.2-7.0;
(2) seed ullage slake is real disappears: sterilization slack tank (sky disappears) earlier, after real jar of the supplementary material sterilization (disappearing in fact) of packing into, with the superheated vapour sterilization, sterilisation temp is 115-126 ℃, tank pressure is 0.11-0.13MPa, the time is 30-40 minute;
(3) inoculation: after the real jar of sterilization,, during to 26-30 ℃, inoculate the seed of bacillus megaterium by the ratio of the 1-2% of supplementary material mass ratio with tube-cooled water cooling material;
(4) seeding tank fermentation: leavening temperature is controlled at 28-30 ℃, the flow that feeds sterile air preceding ten hours be supplementary material volume X (0.4-0.6)/hour, be later on supplementary material volume X (0.9-1.1)/hour, fermentation time is 40-48 hour;
(5) fermentation ullage slake is real disappears: sterilization slack tank earlier, after real jar of the supplementary material sterilization of packing into, with the superheated vapour sterilization, sterilisation temp is 115-126 ℃, tank pressure is 0.11-0.13MPa, the time is 30-40 minute;
(6) inoculation: after real the disappearing, with tube-cooled water cooling material, during to 26-30 ℃, the 8-12% that presses the supplementary material mass ratio inoculates the seed of the bacillus megaterium of seeding tank fermentation gained;
(7) ferment tank: leavening temperature is controlled at 28-30 ℃, the flow that feeds sterile air preceding ten hours be supplementary material volume X (0.4-0.6)/hour, be later on supplementary material volume X (0.9-1.1)/hour, fermentation time is 40-48 hour;
(8) open the bleeder valve discharging and enter storage tank with standby.
Protein and carbohydrate supplementary material are made up of by the starch of 10-12%, the sucrose of 4-6%, the peptone of 1-3%, the yeast extract paste of 0.1-0.3, the ammonium sulfate of 0.8-1.2%, sal epsom and remaining water of 0.4-0.6% mass ratio (down together).Protein that this proportioning is made and the fermentation of carbohydrate supplementary material are very fast, and effect is comparatively desirable.
The production technique of sterile air is: gather air in the upper atmosphere more than the 10M of the face of land, it is carried out coarse filtration remove group's dust, enter gas-holder with the air compression of air compressor machine after with coarse filtration, remove airborne water and oil by gas-liquid separator separates, then air being sent into loft drier further dewaters and cleaningization through cascade filtration, send into heat exchanger again and carry out the constant temperature processing, but then send into the cotton strainer filtration sterilization of constant temperature and pressure autosterilization, but the secondary absolute filter of sending into same constant temperature and pressure autosterilization subsequently filters, detect the quality of sterile air at last with the filter effect determinator, qualified back feeds seeding tank or fermentor tank enters fermentation procedure.The sterile air of this explained hereafter is very clean, and quality is good, can satisfy the requirement of zymotechnique.
Protein and carbohydrate supplementary material are made up of the starch of mass ratio (down together) by 10%, 5% sucrose, 1% peptone, 0.1 yeast extract paste, 1% ammonium sulfate, 0.5% sal epsom and remaining water.Protein that this proportioning is made and the fermentation of carbohydrate supplementary material are fast, and effect is very good.
In operation (3) inoculation, after disappearing in fact, during to 26-30 ℃, press the seed that 2% of supplementary material mass ratio is inoculated bacillus megaterium with tube-cooled water cooling supplementary material; In operation (6) inoculation, after disappearing in fact, with tube-cooled water cooling supplementary material during to 26-30 ℃, the ferment seed of bacillus megaterium of gained of the 10% inoculation seeding tank of press the supplementary material mass ratio.This inoculative proportion is more reasonable, and fermentation is fast, and effect is very good, can not cause the waste of seed again.
This technology is at first poured protein and carbohydrate supplementary material into the mixing of materials jar, and the valve of control water inlet, the valve that advances sodium hydroxide are controlled at 30-35% with the solids concentration of supplementary material, and pH value is 6.2-7.0.Reach after the above-mentioned requirements supplementary material and enter seeding tank and carry out liquid fermenting, whole fermentation process is finished by system.When supplementary material reaches requirement, transmitter is given system with signal feedback, and system sends instruction and regulates to charging and ask that charging is opened between regulating, the beginning charging.High level alarm is installed on seeding tank, and high level alarm and charging switch are asked interlocking.Be that high level alarm is given system with signal feedback, system directive charging switch-valve moves.The charging switch asks automatically and cuts out when arriving high liquid level, and charging stops.
Begin to enter the real operation that disappears of seeding tank after charging finishes, temperature sensor and pressure transmitter are housed on the seeding tank, system CPU is realized sterilization automatically by the parameter of these two transmitters according to default to seeding tank.System's slack tank disinfectant principle is identical with real jar of sterilization before the charging.System opened the inoculation valves by the open and close of the signal control tube-cooled water of temperature sensor automatically when temperature drops to 28 ℃ after sterilization finished, and inoculated.Enter the seeding tank fermenting process.
The flow that feeds sterile air in the fermenting process preceding ten hours be supplementary material volume X (0.4-0.6)/hour, be later on supplementary material volume X (0.9-1.1)/hour, fermentation time is 40-48 hour.The temperature of whole process supplementary material all is by being controlled at 28-30 ℃.The flow of sterile air and time recorder interlocking.
The operating method of fermentor tank is identical with seeding tank, comprises the real sterilization of fermentation ullage slake, inoculation, ferment tank.When the fermentation termination, also be the signal of by automatic turbidity analyser the supplementary material in the jar being analyzed and sent the fermentation termination automatically.After system receives this signal, send the instruction of opening bleeder valve.
The liquid fermenting production unit of bacillus megaterium comprises the worm conveyor (auger) that sodium hydroxide, process water and protein and carbohydrate supplementary material are mixed at least, the mixing of materials jar that is connected with the outlet of worm conveyor, the seeding tank that is connected with the sterile air preparation apparatus with pipeline and mixing of materials jar respectively, the fermentor tank that is connected with the sterile air preparation apparatus with pipeline and seeding tank respectively, the outlet of fermentor tank is connecting bleeder valve, and seeding tank also is connected with steam-pipe with fermentor tank.
The sterile air preparation apparatus comprises the air pump of gathering air in the upper atmosphere more than the 10M of the face of land at least, be connected the coarse filter that air is carried out coarse filtration removal solid dust with the outlet of air pump, the air compressor machine that is connected with coarse filter with pipeline, the gas-holder that is connected with air compressor machine with pipeline, with pipeline and the tank connected gas-liquid separator of gas storage, the loft drier that is connected with gas-liquid separator with pipeline, the secondary filter that is connected with loft drier with pipeline, the heat exchanger that is connected with secondary filter with pipeline, the cotton strainer that is connected with heat exchanger with pipeline, secondary absolute filter that is connected with the cotton strainer with pipeline and the filter effect determinator that is installed in the outlet of secondary absolute filter.
Whole sterile air preparation apparatus adopts full-automatic control.At first adopted the high-altitude gas production, filtered by a coarse filter from the air in the atmosphere, the filtering purpose of this grade is to remove airborne solid dust.Air admission air compressor machine behind the prefiltration enters gas-holder after the air compression.Because coarse filter only can remove a spot of airborne water and oil, thus the gas of gas-holder need by gas-liquid separator with airborne water oil part from, enter loft drier then.
Air in the loft drier passes through heat exchanger again through cascade filtration.Temperature in the heat exchanger can be regulated by the transfer valve of water coolant and steam.Air admission cotton strainer after the processing.On the cotton strainer temp measuring system is installed.When humidity exceeded standard, system can report to the police automatically, and the automatic starting of media processing system meeting of cotton strainer is to guarantee Air quality.
Air then enters the secondary absolute filter.The secondary absolute filter is identical with the cotton strainer, has also realized autosterilization, and its two ends have installed differential manometer additional simultaneously.Be provided with high-low pressure warning dress system mutually by pressure relay and central processing unit, realized real-time monitoring its filtering medium property.
Product of the present invention is liquid, i.e. bacterium liquid.Its available barreled directly dispatches from the factory and sells the core material of producing as microbial fertilizer, perhaps is made into the convenient transportation of strain powder in conjunction with subsequent techniques such as dryings.To dilute earlier during use, be sprayed on the blade face of crop with utensil again and apply fertilizer.Also can be used for the fermentative production of solid fertilizer as seed.
Advantage of the present invention and positively effect are: technical process is reasonable, the most of automatic controls of production unit, has got rid of the living contaminants situation that manual operation causes, and greatly reduces the tank switching rate; The production efficiency height is particularly suitable for producing in enormous quantities; Can monitor at any time bacillus megaterium upgrowth situation, the fermented quality height; Production cost is low.It can be widely used in the production in enormous quantities of bacillus megaterium liquid and solid phase prod.
Embodiment
Embodiment 1: the liquid fermentation process of a kind of bacillus megaterium.It comprises the steps:
(1) batching: protein and carbohydrate supplementary material are made up of the starch of mass ratio (down together) by 10%, 5% sucrose, 1% peptone, 0.1 yeast extract paste, 1% ammonium sulfate, 0.5% sal epsom and remaining water, with protein and carbohydrate supplementary material, water and sodium hydroxide feed material mixing tank, mix while sending, until mixing, the solids concentration that makes supplementary material is 30%, and the pH value of supplementary material is 7.0;
(2) seed ullage slake is real disappears: sterilization slack tank earlier, after real jar of the supplementary material sterilization of packing into, with the superheated vapour sterilization, sterilisation temp is 115 ℃, tank pressure is 0.11MPa, the time is 40 minutes;
(3) inoculation: after real the disappearing,, during to 30 ℃, press the seed of 1% inoculation bacillus megaterium of supplementary material mass ratio with tube-cooled water cooling supplementary material;
(4) seeding tank fermentation: leavening temperature is controlled at 28 ℃, and the flow that feeds sterile air preceding ten hours was supplementary material volume X 0.4/ hour, was later on supplementary material volume X 0.9/ hour, and fermentation time is 40 hours;
(5) fermentation ullage slake is real disappears: sterilization slack tank earlier, after real jar of the supplementary material sterilization of packing into, with the superheated vapour sterilization, sterilisation temp is 115 ℃, tank pressure is 0.11MPa, the time is 40 minutes;
(6) inoculation: after real the disappearing,, during to 30 ℃, press the seed that 8% of supplementary material mass ratio is inoculated the bacillus megaterium of seeding tank fermentation gained with tube-cooled water cooling material.
(7) ferment tank: leavening temperature is controlled at 28 ℃, and the flow that feeds sterile air preceding ten hours was supplementary material volume X 0.4/ hour, was later on supplementary material volume X 0.9/ hour, and fermentation time is 48 hours;
(8) open the bleeder valve discharging and enter storage tank with standby.
The production technique of sterile air is: gather air in the upper atmosphere more than the 10M of the face of land, it is carried out coarse filtration remove group's dust, enter gas-holder with the air compression of air compressor machine after with coarse filtration, remove airborne water and oil by gas-liquid separator separates, then the air admission loft drier is further dewatered and cleaningization through cascade filtration, send into heat exchanger again and carry out the constant temperature processing, but then send into the cotton strainer filtration sterilization of constant temperature and pressure autosterilization, but the secondary absolute filter of sending into same constant temperature and pressure autosterilization subsequently filters, detect the quality of sterile air at last with the filter effect determinator, qualified back feeds seeding tank or fermentor tank enters fermentation procedure.
Embodiment 2: the liquid fermentation process of a kind of bacillus megaterium.It comprises the steps:
(1) batching: protein and carbohydrate supplementary material are made up of the starch of mass ratio (down together) by 12%, 4% sucrose, 2% peptone, 0.2 yeast extract paste, 0.8% ammonium sulfate, 0.6% sal epsom and remaining water, with protein and carbohydrate supplementary material, water and sodium hydroxide feed material mixing tank, mix while sending, until mixing, the solids concentration that makes supplementary material is 35%, and the pH value of supplementary material is 6.2;
(2) seed ullage slake is real disappears: sterilization slack tank earlier, after real jar of the supplementary material sterilization of packing into, with the superheated vapour sterilization, sterilisation temp is 126 ℃, tank pressure is 0.13MPa, the time is 30 minutes;
(3) inoculation: after real the disappearing,, during to 26 ℃, press the seed of 2% inoculation bacillus megaterium of supplementary material mass ratio with tube-cooled water cooling supplementary material;
(4) seeding tank fermentation: leavening temperature is controlled at 30 ℃, and the flow that feeds sterile air preceding ten hours was supplementary material volume X 0.6/ hour, was later on supplementary material volume X 1.1/ hour, and fermentation time is 46 hours;
(5) fermentation ullage slake is real disappears: sterilization slack tank earlier, after real jar of the supplementary material sterilization of packing into, with the superheated vapour sterilization, sterilisation temp is 120 ℃, tank pressure is 0.13MPa, the time is 35 minutes;
(6) inoculation: after real the disappearing,, during to 28 ℃, press the seed that 12% of supplementary material mass ratio is inoculated the bacillus megaterium of seeding tank fermentation gained with tube-cooled water cooling material;
(7) ferment tank: leavening temperature is controlled at 30 ℃, and the flow that feeds sterile air preceding ten hours was supplementary material volume X 0.5/ hour, was later on supplementary material volume X 1.1/ hour, and fermentation time is 45 hours;
(8) open the bleeder valve discharging and enter storage tank with standby.
The production technique of sterile air is: gather air in the upper atmosphere more than the 10M of the face of land, it is carried out coarse filtration remove group's dust, enter gas-holder with the air compression of air compressor machine after with coarse filtration, remove airborne water and oil by gas-liquid separator separates, then the air admission loft drier is further dewatered and cleaningization through cascade filtration, send into heat exchanger again and carry out the constant temperature processing, but then send into the cotton strainer filtration sterilization of constant temperature and pressure autosterilization, but the secondary absolute filter of sending into same constant temperature and pressure autosterilization subsequently filters, detect the quality of sterile air at last with the filter effect determinator, qualified back feeds seeding tank or fermentor tank enters fermentation procedure.
Embodiment 3: the liquid fermentation process of a kind of bacillus megaterium.It comprises the steps:
(1) batching: protein and carbohydrate supplementary material are made up of by 10% starch, 6% sucrose, 3% peptone, 0.3 yeast extract paste, 1.2% ammonium sulfate, 0.4% sal epsom and remaining water mass ratio, with protein and carbohydrate supplementary material, water and sodium hydroxide feed material mixing tank, mix while sending, until mixing, the solids concentration that makes supplementary material is 35%, and the pH value of supplementary material is 6.2;
(2) seed ullage slake is real disappears: sterilization slack tank earlier, after real jar of the supplementary material sterilization of packing into, with the superheated vapour sterilization, sterilisation temp is 120 ℃, tank pressure is 0.12MPa, the time is 35 minutes;
(3) inoculation: after real the disappearing,, during to 28 ℃, press the seed of 1.5% inoculation bacillus megaterium of supplementary material mass ratio with tube-cooled water cooling supplementary material;
(4) seeding tank fermentation: leavening temperature is controlled at 30 ℃, and the flow that feeds sterile air preceding ten hours was supplementary material volume X 0.5/ hour, was later on supplementary material volume X 1.0/ hour, and fermentation time is 48 hours;
(5) fermentation ullage slake is real disappears: sterilization slack tank earlier, after real jar of the supplementary material sterilization of packing into, with the superheated vapour sterilization, sterilisation temp is 126 ℃, tank pressure is 0.12MPa, the time is 30 minutes;
(6) inoculation: after real the disappearing,, during to 26 ℃, press the seed that 10% of supplementary material mass ratio is inoculated the bacillus megaterium of seeding tank fermentation gained with tube-cooled water cooling material;
(7) ferment tank: leavening temperature is controlled at 30 ℃, and the flow that feeds sterile air preceding ten hours was supplementary material volume X 0.6/ hour, was later on supplementary material volume X 1.0/ hour, and fermentation time is 40 hours;
(8) open the bleeder valve discharging and enter storage tank with standby.
The production technique of sterile air is: gather air in the upper atmosphere more than the 10M of the face of land, it is carried out coarse filtration remove group's dust, enter gas-holder with the air compression of air compressor machine after with coarse filtration, remove airborne water and oil by gas-liquid separator separates, then the air admission loft drier is further dewatered and cleaningization through cascade filtration, send into heat exchanger again and carry out the constant temperature processing, but then send into the cotton strainer filtration sterilization of constant temperature and pressure autosterilization, but the secondary absolute filter of sending into same constant temperature and pressure autosterilization subsequently filters, detect the quality of sterile air at last with the filter effect determinator, qualified back feeds seeding tank or fermentor tank enters fermentation procedure.
Claims (5)
1. the liquid fermentation process of a bacillus megaterium is characterized in that: comprise the steps:
(1), batching: protein and carbohydrate supplementary material are made up of by the starch of 10-12%, the sucrose of 4-6%, the peptone of 1-3%, the yeast extract paste of 0.1-0.3, the ammonium sulfate of 0.8-1.2%, sal epsom and remaining water of 0.4-0.6% mass ratio, with protein and carbohydrate supplementary material, water and sodium hydroxide feed material mixing tank, mix while sending, until mixing, the solids concentration that makes supplementary material is 30-35%, and the pH value of supplementary material is 6.2-7.0;
(2), seed ullage slake is real disappears: sterilization slack tank earlier, after real jar of the supplementary material sterilization of packing into, with the superheated vapour sterilization, sterilisation temp is 115-126 ℃, tank pressure is 0.11-0.13MPa, the time is 30-40 minute;
(3), inoculation: after real the disappearing,, during to 26-30 ℃, inoculate the seed of bacillus megaterium by the ratio of the 1-2% of supplementary material mass ratio with tube-cooled water cooling material;
(4), seeding tank fermentation: leavening temperature is controlled at 28-30 ℃, the flow that feeds sterile air preceding ten hours be supplementary material volume X (0.4～0.6)/hour, be later on supplementary material volume X (0.9-1.1)/hour, fermentation time is 40-48 hour;
(5), fermentation ullage slake is real disappears: sterilization slack tank earlier, after real jar of the supplementary material sterilization of packing into, with the superheated vapour sterilization, sterilisation temp is 115-126 ℃, tank pressure is 0.11-0.13MPa, the time is 30-40 minute;
(6), inoculation: after real the disappearing, with tube-cooled water cooling material, during to 26-30 ℃, the 8-12% that presses the supplementary material mass ratio inoculates the seed of the bacillus megaterium of seeding tank fermentation gained;
(7), ferment tank: leavening temperature is controlled at 28-30 ℃, the flow that feeds sterile air preceding ten hours be supplementary material volume X (0.4-0.6)/hour, be later on supplementary material volume X (0.9-1.1)/hour, fermentation time is 40-48 hour;
(8), open the bleeder valve discharging and enter storage tank with standby.
2. according to the liquid fermentation process of the described bacillus megaterium of claim 1, it is characterized in that: the production technique of described sterile air is: gather air in the upper atmosphere more than the 10M of the face of land, it is carried out coarse filtration remove group's dust, enter gas-holder with the air compression of air compressor machine after with coarse filtration, remove airborne water and oil by gas-liquid separator separates, then air being sent into loft drier further dewaters and cleaningization through cascade filtration, send into heat exchanger again and carry out the constant temperature processing, but then send into the cotton strainer filtration sterilization of constant temperature and pressure autosterilization, but the secondary absolute filter of sending into same constant temperature and pressure autosterilization subsequently filters, detect the quality of sterile air at last with the filter effect determinator, qualified back feeds seeding tank or fermentor tank enters fermentation procedure.
3. according to the liquid fermentation process of claim 1 or 2 described bacillus megateriums, it is characterized in that: described protein and carbohydrate supplementary material are made up of by 10% starch, 5% sucrose, 1% peptone, 0.1 yeast extract paste, 1% ammonium sulfate, 0.5% sal epsom and remaining water mass ratio.
4. according to the liquid fermentation process of claim 1 or 2 described bacillus megateriums, it is characterized in that: in described operation (3) inoculation, after disappearing in fact, during to 26-30% ℃, press the seed that 2% of supplementary material mass ratio is inoculated bacillus megaterium with tube-cooled water cooling material.
5. according to the liquid fermentation process of claim 1 or 2 described bacillus megateriums, it is characterized in that: in described operation (6) inoculation, after disappearing in fact, with tube-cooled water cooling supplementary material during, press the seed of bacillus megaterium of 10% inoculation seeding tank fermentation gained of supplementary material mass ratio to 26-30 ℃.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103570434A (en) * | 2012-07-20 | 2014-02-12 | 上海绿乐生物科技有限公司 | Microbial liquid, liquid microbial fertilizer, and production method and applications thereof |
| CN115386522A (en) * | 2022-09-30 | 2022-11-25 | 山东天润和生物工程有限公司 | Fermentation method of clostridium butyricum |
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2009
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103570434A (en) * | 2012-07-20 | 2014-02-12 | 上海绿乐生物科技有限公司 | Microbial liquid, liquid microbial fertilizer, and production method and applications thereof |
| CN103570434B (en) * | 2012-07-20 | 2016-01-13 | 上海绿乐生物科技有限公司 | A kind of microbial inoculum, liquid microbe fertilizer, its production method and application |
| CN115386522A (en) * | 2022-09-30 | 2022-11-25 | 山东天润和生物工程有限公司 | Fermentation method of clostridium butyricum |
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