CN102154091A - Fermentation tank for bacillus thuringiensis to generate unit cell - Google Patents
Fermentation tank for bacillus thuringiensis to generate unit cell Download PDFInfo
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- CN102154091A CN102154091A CN2010106237421A CN201010623742A CN102154091A CN 102154091 A CN102154091 A CN 102154091A CN 2010106237421 A CN2010106237421 A CN 2010106237421A CN 201010623742 A CN201010623742 A CN 201010623742A CN 102154091 A CN102154091 A CN 102154091A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/02—Stirrer or mobile mixing elements
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/22—Settling tanks; Sedimentation by gravity
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
- C12M41/22—Heat exchange systems, e.g. heat jackets or outer envelopes in contact with the bioreactor walls
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Abstract
The invention provides a fermentation tank for bacillus thuringiensis to generate unit cells, the height-diameter ratio is from 2:1 to 3:1, and the cone bottom and the tank top are provided with ellipse heads; 2-4 jet flow mixers are arranged above the cone bottom of a tank body, the cone bottom of the tank body is provide with one cooling belt, and two cooling belts are arranged at the 1/3-2/3 above the cone bottom of the tank body; the tank top is provided with a safety valve, a vacuum valve, a vapor-liquid shunt interface and an overhaul inlet, the cone bottom of the tank body is provided with one discharge hole, and one discharge hole is respectively arranged under the two cooling belts at the upper part of the tank body; and the fermentation tank is matched with a normal fermentation tank to be specially used for performing the post-fermentation on fermentation liquid when germ is cultivated to form at first in the normal fermentation tank. Therefore, the energy consumption caused by the fermentation can be reduced, the yield of the germ and crystalloid can be improved, and the fermentation tank is good for separation of the crystalloid and the germ.
Description
Technical field
The invention belongs to the fermentation equipment field, particularly relate to Tribactur (Bt) the fermentation later stage product brood cell and the fermentation equipment in crystal stage, promptly Tribactur produces the structure cell fermentor tank.
Background technology
Oxygen is difficult to water-soluble generally speaking, in fermented liquid, contain other various compositions, the solubleness of oxygen is than lower in pure water, and the high more dissolved oxygen speed of the concentration of fermented liquid is slow more, and the oxygen that needs in microbial respiratory and the Metabolic activity must be dissolved in and could directly be utilized in the fermented liquid, and the deficiency of oxygen supply and the briefly interrupted of oxygen supply will cause serious irreversible lesion to fermentation.Therefore, design fermentor tank effectively and reasonably, be significant.In recent years, domestic and international dissolved oxygen and the power saving of investigator in order to solve the ventilating fermentation process, the ventilation fermentation tank kind that proposes is a lot, developed some novel fermentation jars such as air lift type, self-priming, jet-type. one of main development tendency is to carry out the transition to air-flow from mechanical stirring to stir, and improved core is to improve oxygen transfer coefficient and conserve energy.
Mechanical agitating fermentation tank typical case representative is general form mechanical agitating fermentation tank and self-suction fermentor.Gas is subjected to twice dispersion in liquid phase in this class jar, and the capacity usage ratio of this jar of gas dispersion is lower.Self-suction fermentor adopts negative pressure fermentation, can obtain more than high Kla value (the liquid phase volume oxygen mass transfer coefficient of airlift fermentor, unit: L/h), but can not implement the air pressurize, in case running into equipment failure stirs and to be obstructed or situation such as power failure. will cause thalline death by suffocation, make stuck fermentation, even cause tank switching.The domestic and international at present generally Bt fermentation reactor of usefulness all is the mechanical agitation type fermentor tank, the most frequently used production method of Bt is to stir the fermentative production of finishing Bt in the ventilation fermentation tank at one, general six curved leaf turbine type and the arrow leaf formulas that adopt of its agitator, internal structure is complicated to have drive disk assembly with an axle envelope place, making has the dead angle and is difficult to realize sealing in the jar, assorted bacterium of easy infection and phage, cause the fermentation failure, and be subjected to structural limitations, the tank body aspect ratio is little, can not utilize the oxygen in the sterile air, thereby the sterile air requirement is big morely, jar cost height, energy consumption is big, has increased production cost.Data statistics shows, is used for mechanical type and stirs the energy that is consumed and account for about half of fermentation whole process.Continuous use along with superior strain, high density fermentation is depended merely on increases ventilation and stirring velocity, the supply of oxygen is limited, and the fermentation of Bt is divided into the yeast culture stage and brood cell, crystallin generation come off the stage, the purpose of producing is in order to obtain maximum have active brood cell of virulence and crystallins, in the later stage of Bt fermentation, intensive mechanical stirring, thalline, brood cell and crystal are damaged, be unfavorable for the formation of Pesticidal toxins.
The air-flow stirred fermentor mainly cooperates by nozzle and inside and outside circulating pipe or sieve plate disperses gas, mechanical rotating part, and typical case's representative is an airlift fermentor.Some investigators improve the power ventilation fermentor tank of producing Bt, replace mechanical agitating fermentation tank fermentative production Bt as adopting annular space gas lifting type loop reactor.The air that airlift fermentor feeds makes a jar interior different zones form density difference and macroscopical cycling stream, form local turbulence and microcosmic mixed flow simultaneously, to reach stirring, mix and to make the purpose of airborne oxygen dissolution in fermented liquid, its advantage be have simple in structure, movement-less part, aseptic technique reliability height, power consumption is few, operating maintenance is convenient and cost is low, higher fermentor tank height is through than (H/D), as height through than 1: 5-1: 8, can increase the duration of contact of oxygen and feed liquid, increase the solubleness of oxygen in feed liquid; Facts have proved that when the watt consumption of ventilation and unit volume was certain, H/D was that 2 drafting efficiency is that H/D is 1 1.4 times.The fermentation of employing airlift fermentor can make the fermentation time of Bt shorten, and the bacterium number improves.When airlift fermentor is used for hanging down the fermentation of solid content substratum, excellent property, but can not be used for the fermentation of high solid content substratum, the oxygen delivery capacity of reactor remains further to be improved.Have and settle some deflection elements, netted airlift fermentor in the guide shell that is reported in gas lift type common loop reactor, performance is improved, but it is limited that fermentor tank amplifies, compare need with mechanical stirring and increase ventilation, as dispose the outside circulation pipe and increase dissolved oxygen, the then outside pumping and the recycle system easily make fermented liquid be subjected to living contaminants and make cell injury, and too high aspect ratio makes the temperature mixing difficult, and the precipitation of cell, albumen etc. is also difficult.
The problem that the breather that traditional general form fermentor tank is adopted exists: bubble diameter increases along with the increase of air flow, and bobble rise velocity also increases thereupon; The bubble degree of grinding does not but increase along with the increase of mixing speed, it can only maintain on the same order of magnitude, increasing air flow and increasing power of agitator not only can not have by a relatively large margin raising to the container mass transfer coefficient, will cause the consumption of a large amount of energy and the raising of product cost on the contrary.Strengthen gas dispersion by the jet mixer that the investigator adopts nozzle, mixing tube and circulation tube to form, reduce bubble diameter, help the raising of increase, the growth of duration of contact and the volumetric mass transfer coefficient of gas, liquid two-phase specific surface area.Production practice show, with jet mixer the number of plies of agitator are reduced, and make the mechanical stirring ventilation fermentation tank improve dissolved oxygen effect, energy efficient more than 20%.
Fermentative production Bt product, what need is crystal, brood cell and the supernatant liquor of Bt.Present fermentor tank generally all is a round bottom, and tank body does not adopt the multistage cooling, does not have the segmentation discharging yet, can't promote precipitation to fermented liquid segmentation cooling, and by the discharging respectively of its different concns, but by the discharging together of jar bottom bleeder valve, filtration and centrifuging process cycle are long, and energy consumption is also big.
Summary of the invention
The purpose of this invention is to provide a kind of fermentor tank that Tribactur produces the structure cell stage that is used for,, improve brood cell, crystalline output, help separating of crystal and brood cell simultaneously to reduce the energy consumption of fermenting process.
The present invention realizes that as follows a kind of Tribactur produces the fermentor tank in structure cell stage, its aspect ratio 2: 1 to 3: 1, the awl end, tank deck elipse head; Top, the tank body awl end is equipped with 2-4 jet mixer, and the 1/3-2/3 place that 1 cooling zone, top, the tank body awl end are established in the tank body awl end establishes 2 cooling zones; Tank deck is provided with safety valve, vacuum valve, vapour-liquid splitter interface, maintenance hand-hole, and tank body awl bottom is provided with the below of a discharge port, 2 cooling zones in tank body top and respectively establishes 1 discharge port.
Above-mentioned a kind of Tribactur produces the fermentor tank in structure cell stage, and awl base angle degree is 70 °, establishes CIP original position washing unit in the tank body.
A kind of Tribactur provided by the present invention produces the front end of the fermentor tank in structure cell stage, can select mechanical stirring or airlift fermentor to be used for yeast culture, promptly as medium feed, be seeded to the brood cell and just formed the stage, or claim the fermentor tank in primary fermentation stage; And a kind of Tribactur of the present invention produces the fermentor tank in structure cell stage, is used for crystal, brood cell's formation, and promptly later stage fermentation usefulness can reduce the stirring energy of ventilation fermentation tank or the ventilation of airlift fermentor, also is equivalent to the effect of transfer jar.Its ferment effect and benefit are fairly obvious:
(1) 2-4 jet mixer is installed with enhancing gaseous exchange and dispersion, and can be had more dissolved oxygen requirement selection unlatching jet mixer size and number, be convenient to the control of oxygen.No whipping appts, the weakness that can overcome mechanical agitating fermentation tank is better than airlift fermentor stirring and dissolved oxygen again.
(2) aspect ratio between mechanical stirring ventilation fermentation tank and airlift fermentor (2: 1 to 3: 1) can prolong the duration of contact of air and feed liquid, increases the solubleness of oxygen in feed liquid, and makes whole jar of fermented liquid have good convection current mixed effect.
(3) fermentor tank upwards is respectively equipped with 3 sections cooling zones from the awl end, rational temperature control can make fermented liquid form systemic circulation and countless partial circulating, promote the dissolving of oxygen and the mixing of temperature, make the temperature of whole fermented liquid even, help convection current and the heat exchange of fermented liquid in jar.This fermentor tank mixing principle except turbulent flow and microcosmic mixed flow that the mixing of airlift fermentor forms, the convection current that also has the variable density of the stirring action that the jet mixer air inlet forms, cold and hot action of thermal difference, liquids and gases to cause.
(4) be provided with three discharge ports, discharging from top to bottom successively.The segmentation discharging can obtain the fermented liquid of different opacities, significantly reduces the filtration latch up phenomenon by clear to the dense aftertreatment of carrying out fermented liquid, improves filtration velocity.The denseer fermented liquid that discharge the bottom can be directly used in dry or centrifugal, reduces the treatment capacity of later stage fermentation liquid, shortens the aftertreatment time, reduces production costs.
(5) awl is provided with cooling zone in the end, after the later stage leaves standstill or adds pre-treatment fermented liquid such as flocculation agent, help big material such as crystal, brood cell precipitation, be collected in conical lower portion.At the bottom of the fermentor tank for the awl end, can reduce cylindrical can at the bottom of elipse head sedimentary residual.Awl base angle degree is 70 °, can reduce the eliminating resistance, is convenient to sedimental discharge.
In addition, tank deck adopts elipse head, at tank deck safety valve, vacuum valve, vapour-liquid splitter interface is set, and supplied gas is discharged and the fermentor tank scavenging solution enters, and establishes hand-hole, uses for maintenance; If CIP original position washing unit, lamp mirror and visor; Tall and thin coned fermentation tank develops to facade, has saved the place.
Description of drawings
The present invention is further described below in conjunction with accompanying drawing
A kind of Tribactur of Fig. 1 produces the fermentor tank in structure cell stage
1, cooling zone; 2, sterile air inlet pipe; 3, material feed-pipe; 4, vapor pipe;
5, discharge port; 6, CIP original position washing unit; 7, jet mixer.
Specific embodiment
Embodiment 1
Bacterial strain: with 8010 is fermentation strain fermentative production Bt.
Equipment: cultivate charging, be seeded to firm product born of the same parents' stage, adopt traditional 1.5L power ventilation stirred pot; Tribactur produces the fermentation stage in structure cell stage, adopt 1.5L a kind of Tribactur as shown in Figure 1 to produce the fermentor tank in structure cell stage, its aspect ratio 2: 1,70 ° of awl ends, the tank deck elipse head, top, the tank body awl end is equipped with 2 jet mixers, the tank body awl end and top 1/3 thereof and 2/3 eminence are respectively established 1 cooling zone, be provided with safety valve, vacuum valve, vapour-liquid splitter interface, maintenance hand-hole at tank deck, tank body awl bottom is provided with a discharge port, top, the awl end 1/4 and 1/2 locate respectively establish 1 discharge port.
Culture medium prescription: W-Gum 1%, dregs of beans 1.3%, groundnut meal 1%, corn steep liquor 1%, yeast powder 0.1%, CaCO
30.3%, KH
2PO
40.2%, MgSO
40.07%, pH 7.0-7.5,121 ℃ of autoclaving 40min.
Fermented liquid brood cell method of counting: get 85 ℃ of heat treated 5min of fermented liquid that fermentation has just finished, dilution plate viable bacteria technical process counting is adopted in cooling rapidly;
The toxicity test method: fermented liquid after 1000 times of dilutions to the biological assay of 2-3 small cabbage moth in age.
Method: medium feed, be seeded to firm product born of the same parents' stage and in 1.5L power ventilation stirred pot, ferment, behind the culture medium inoculated of sterilization, be controlled at 30 ℃ by substratum and sterile air amount 1: 1.5v/v/min, leavening temperature, the fermentation condition of mixing speed 250r/min, tank pressure 0.5atm, in 1.5L power ventilation stirred pot, ferment, when being cultured to the brood cell and just having begun to form, the fermented liquid of its generation claims primary fermentation liquid:
Primary fermentation liquid changes a kind of Tribactur provided by the present invention over to and produces the fermentor tank in structure cell stage, control sterile air amount 1: 1v/v/min, temperature is controlled at 33 ℃ of fermentations, ferment when the brood cell has just begun to come off, stop to ventilate, keep 33 ℃ when the brood cell comes off more than 50%, transfer about pH to 5, add flocculation agent etc., discharging changes the post-processed stage over to.
The result: fermentation time is 60h, the several 9,500,000,000/mL of finished product brood cell, and brood cell's sync rates is more than 95%, and crystal is neat; 48h small cabbage moth corrected mortality 100%; And traditional single batch fermentation scheme, fermentation time need be 71h, the several 6,100,000,000/mL of the brood cell of finished product, brood cell's rate of formation about 80%, 48h 2-3 small cabbage moth in age corrected mortality 83%.By 0.73 yuan of every degree electricity, every jar of 1.5t calculates, and can save and stir energy consumption more than 300 yuan, and fermentation period shortens 11h, and fermented liquid brood cell number has increased by 3,400,000,000/mL, and the small cabbage moth killing ability is improved 20%.The later stage membrane filtration is accelerated, and can shift to an earlier date 1h and finish post-processing stages.
Embodiment 2
As embodiment 1, medium feed, be seeded to firm product born of the same parents' stage, adopt traditional 2.5L power ventilation stirred pot; Tribactur produces the fermentation stage in structure cell stage, adopt 2.5L a kind of Tribactur as shown in Figure 1 to produce the fermentor tank in structure cell stage, its aspect ratio 2.5: 1, the awl end, the tank deck elipse head, top, the tank body awl end is equipped with 4 jet mixers, the tank body awl end and top 1/3 thereof and 3/5 eminence are respectively established 1 cooling zone, be provided with safety valve, vacuum valve, vapour-liquid splitter interface, maintenance hand-hole at tank deck, 1/4 and 1/2 place that tank body awl bottom is provided with a discharge port, top, the awl end respectively establishes 1 discharge port.
The effect of product is with embodiment 1, and benefit is slightly higher than embodiment 1.
Claims (2)
1. a Tribactur produces the fermentor tank in structure cell stage, it is characterized in that the fermentor tank aspect ratio 2: 1 to 3: 1, the awl end, tank deck elipse head; Top, the tank body awl end is equipped with 2-4 jet mixer, and the 1/3-2/3 place that 1 cooling zone, top, the tank body awl end are established in the tank body awl end establishes 2 cooling zones; Tank deck is provided with safety valve, vacuum valve, vapour-liquid splitter interface, maintenance hand-hole, and tank body awl bottom is provided with the below of a discharge port, 2 cooling zones in tank body top and respectively establishes 1 discharge port.
2. a kind of Tribactur according to claim 1 produces the fermentor tank in structure cell stage, and it is characterized in that boring the base angle degree is 70 °, establishes CIP original position washing unit in the tank body.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010623742 CN102154091B (en) | 2010-12-25 | 2010-12-25 | Fermentation tank for bacillus thuringiensis to generate unit cell |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010623742 CN102154091B (en) | 2010-12-25 | 2010-12-25 | Fermentation tank for bacillus thuringiensis to generate unit cell |
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| Publication Number | Publication Date |
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| CN102154091A true CN102154091A (en) | 2011-08-17 |
| CN102154091B CN102154091B (en) | 2013-10-02 |
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| CN 201010623742 Expired - Fee Related CN102154091B (en) | 2010-12-25 | 2010-12-25 | Fermentation tank for bacillus thuringiensis to generate unit cell |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112481114A (en) * | 2020-11-30 | 2021-03-12 | 中商构能生态科技(天津)有限公司 | Liquid fermentation system for preparing broussonetia papyrifera fermentation liquor and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN87216445U (en) * | 1987-12-17 | 1988-09-21 | 浙江省石油化工设计院 | Jet-type aerobic fermentation pot |
| CN1033528A (en) * | 1987-12-17 | 1989-06-28 | 浙江省石油化工设计院 | Jet-type aerobic leavening tank |
| CN2251557Y (en) * | 1995-11-24 | 1997-04-09 | 云南省轻纺工业设计院 | Combined high efficiency anaerobic fermentation tank |
-
2010
- 2010-12-25 CN CN 201010623742 patent/CN102154091B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN87216445U (en) * | 1987-12-17 | 1988-09-21 | 浙江省石油化工设计院 | Jet-type aerobic fermentation pot |
| CN1033528A (en) * | 1987-12-17 | 1989-06-28 | 浙江省石油化工设计院 | Jet-type aerobic leavening tank |
| CN2251557Y (en) * | 1995-11-24 | 1997-04-09 | 云南省轻纺工业设计院 | Combined high efficiency anaerobic fermentation tank |
Non-Patent Citations (2)
| Title |
|---|
| F.R.SCHMIDT: "Optimization and scale up of industrial fermentation processes", 《APPL MICROBIOL BIOTECHNOL》, vol. 68, no. 4, 31 December 2005 (2005-12-31), pages 425 - 435 * |
| 徐清华等: "机械搅拌通风发酵罐的节能设计", 《广西轻工业》, vol. 2009, no. 2, 28 February 2009 (2009-02-28) * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112481114A (en) * | 2020-11-30 | 2021-03-12 | 中商构能生态科技(天津)有限公司 | Liquid fermentation system for preparing broussonetia papyrifera fermentation liquor and application thereof |
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