CN113801774A - Thallus culture equipment and method - Google Patents
Thallus culture equipment and method Download PDFInfo
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- CN113801774A CN113801774A CN202111114562.5A CN202111114562A CN113801774A CN 113801774 A CN113801774 A CN 113801774A CN 202111114562 A CN202111114562 A CN 202111114562A CN 113801774 A CN113801774 A CN 113801774A
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- 238000000034 method Methods 0.000 title claims abstract description 16
- 238000000855 fermentation Methods 0.000 claims abstract description 94
- 230000004151 fermentation Effects 0.000 claims abstract description 94
- 239000000463 material Substances 0.000 claims abstract description 47
- 239000007788 liquid Substances 0.000 claims abstract description 34
- 238000004062 sedimentation Methods 0.000 claims abstract description 24
- 238000000926 separation method Methods 0.000 claims abstract description 23
- 238000001514 detection method Methods 0.000 claims abstract description 17
- 238000007599 discharging Methods 0.000 claims abstract description 13
- 238000003756 stirring Methods 0.000 claims description 100
- 241001052560 Thallis Species 0.000 claims description 27
- 238000012258 culturing Methods 0.000 claims description 18
- 230000001580 bacterial effect Effects 0.000 claims description 16
- 239000001963 growth medium Substances 0.000 claims description 9
- 230000003698 anagen phase Effects 0.000 claims description 3
- 230000005540 biological transmission Effects 0.000 claims description 3
- 238000004113 cell culture Methods 0.000 claims 1
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- 239000012530 fluid Substances 0.000 abstract description 6
- 238000011897 real-time detection Methods 0.000 abstract description 3
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 6
- 240000006024 Lactobacillus plantarum Species 0.000 description 6
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 6
- 239000008103 glucose Substances 0.000 description 6
- 229940072205 lactobacillus plantarum Drugs 0.000 description 6
- 239000000758 substrate Substances 0.000 description 6
- 238000012136 culture method Methods 0.000 description 5
- 238000011081 inoculation Methods 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 230000012010 growth Effects 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 241000186660 Lactobacillus Species 0.000 description 3
- 241000235033 Zygosaccharomyces rouxii Species 0.000 description 3
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Abstract
The invention belongs to the technical field of thallus culture, and particularly discloses a thallus culture device and a method, aiming at solving the problems that the existing thallus culture device is complex in structure and is not beneficial to timely separation of thallus. According to the thallus culture equipment, the first chamber, the second chamber and the third chamber which are sequentially connected from top to bottom are arranged in the fermentation tank, thallus obtained by fermentation in the first chamber can be enriched by using the second chamber, and the thallus enriched in the second chamber can be fully accumulated and settled by using the third chamber, so that the thallus can be concentrated without additionally arranging devices such as a rotary sedimentation tank, a circulating pump, a circulating pipe and the like, and the equipment structure is simplified; simultaneously, through setting up turbidity detection device in the third chamber, do benefit to the turbidity of liquid in the real-time detection third chamber to be connected through vertical pipeline with centrifugal device's inlet and the leakage fluid dram of third chamber lower extreme, do benefit to in time discharging the material to centrifugal device after satisfying the turbidity requirement, in order to obtain the active mud of thallus with the quick centrifugal separation of material.
Description
Technical Field
The invention belongs to the technical field of thallus culture, and particularly relates to a thallus culture device and method.
Background
The thallus culture equipment is a facility equipment for providing a suitable environment for thallus growth according to the biological fermentation principle. At present, people adopt more high-density culture methods, namely, culture conditions are optimized, nutrients and oxygen are supplemented, or inhibition factors are removed, so that the quantity of thalli in single-batch fermentation is increased, and higher substrate utilization efficiency or product yield is obtained.
Because the thalli is separated from the fermentation tank in time in the fermentation process, the excessive consumption of nutrient substances and the generation of wastes can be reduced, the growth time of the thalli is prolonged, and the high-efficiency fermentation effect is obtained, the thalli culture equipment usually comprises a centrifugal device besides the fermentation tank and a stirring device. For example: the patent of Chinese utility model with the publication number of CN213624107U discloses a yeast fermentation separation device, which comprises a fermentation tank and a rotary sedimentation tank, wherein the fermentation tank is connected with the rotary sedimentation tank through a circulating pipe, a centrifugal pump and a rotary sedimentation feed valve are respectively arranged on the circulating pipe, an air inlet pipe and a fermentation return pipe are connected on the circulating pipe between the centrifugal pump and the rotary sedimentation feed valve, the other end of the fermentation return pipe is connected with the upper part of the fermentation tank, a centrifugal clear liquid pipe and a rotary sedimentation return pipe are connected on the fermentation return pipe between the fermentation return valve and the fermentation tank, the other end of the centrifugal clear liquid pipe is connected with a solid-liquid centrifuge, and the other end of the rotary sedimentation return pipe is connected with the rotary sedimentation tank; the bottom of the rotary sedimentation tank is connected with a solid-liquid centrifuge through a sedimentation material valve, and the bottom of the solid-liquid centrifuge is provided with a concentration yeast valve.
Although the yeast fermentation separation device can fully utilize the operation of the centrifugal pump to drive the fermentation liquor to be pre-separated in the rotary sedimentation tank, the operation time of the centrifugal machine is greatly reduced, thereby saving the operation cost, reducing the production cost of the yeast and simultaneously improving the fermentation effect; but it has increased and has had rotary sedimentation jar, centrifugal pump and circulating pipe, and when separating the thallus, need with the zymotic fluid pump in the rotary sedimentation jar, the zymotic fluid is discharged the deposit behind the rotary sedimentation jar and is made the supernatant fluid backward flow go into the fermentation cylinder when centrifuge, and not only device structure is complicated, and the thallus is cultivateed and is separated separately to be gone on moreover, can't realize the online separation of thallus, leads to the thallus to separate untimely, is unfavorable for the active maintenance of thallus.
Disclosure of Invention
The invention provides a thallus culture device, and aims to solve the problems that the existing thallus culture device is complex in structure and is not beneficial to timely separation of thallus.
The technical scheme adopted by the invention for solving the technical problems is as follows: the thallus culture equipment comprises a fermentation tank, a stirring device, a centrifugal device and a turbidity detection device; the fermentation tank is provided with a first chamber, a second chamber and a third chamber which are sequentially connected from top to bottom, and the lower end of the third chamber is provided with a liquid outlet; the stirring device is arranged on the fermentation tank, and stirring paddles of the stirring device are positioned in the first chamber and the second chamber; the turbidity detection device is arranged in the third chamber, and a liquid inlet of the centrifugal device is connected with a liquid outlet through a vertical pipeline.
Further, the stirring device comprises a stirring shaft vertically arranged on the fermentation tank and extending into the first chamber and the second chamber, a stirring blade arranged on the stirring shaft, and a stirring motor arranged on the fermentation tank and in transmission connection with the stirring shaft; the stirring shaft and the stirring blade jointly form a stirring paddle of the stirring device.
Further, a valve is arranged on the vertical pipeline.
Further, the centrifugal device is a disk centrifuge.
Further, the volume of the first chamber is 65-75% of the total volume of the fermentation tank, the volume of the second chamber is 20-30% of the total volume of the fermentation tank, and the volume of the third chamber is 2-5% of the total volume of the fermentation tank.
Further, the first chamber comprises a cylindrical main body and an inverted cone-shaped bottom arranged at the lower end of the cylindrical main body, the second chamber and the third chamber are in inverted cone-shaped structures, and the first chamber, the second chamber and the third chamber are coaxially arranged.
Further, the diameter of the joint of the first chamber and the second chamber is 40-50% of the diameter of the cylindrical main body of the first chamber, and the diameter of the joint of the second chamber and the third chamber is 30-40% of the diameter of the upper end of the third chamber.
Further, the taper of the third chamber is 0.7: 1-1.2: 1.
The invention also provides a thallus culture method capable of improving the utilization rate of the substrate and the yield of the product, and the method adopts the thallus culture equipment to produce thallus.
Further, the above method comprises a fermentation step, a sedimentation step and a separation step;
a fermentation step: adding the sterilized culture medium into a fermentation tank through a first chamber, adding the thallus seed liquid into the fermentation tank through the first chamber, and starting a stirring device to stir and ferment; a settling step: stopping the stirring device after the fermentation of the thallus seeds reaches the logarithmic growth phase; after 3-8 min, starting the stirring device to stir at the rotating speed of 2-10 rpm for 0.5-2 min, so that part of thalli are settled into a third chamber; then, continuing stirring and fermenting according to the stirring mode of the fermentation step;
a separation step: performing cyclic operation according to the sedimentation step, and discharging materials to the centrifugal device through a vertical pipeline after the turbidity detection device detects that the turbidity of the liquid in the third chamber exceeds a set value, wherein the volume of the discharged materials is 80-150% of the volume of the third chamber; after the materials are discharged, the culture solution is supplemented into the first chamber, and the volume of the supplemented culture solution is equal to the volume of the discharged materials; discharging the materials into a centrifugal device, and carrying out centrifugal separation on the materials in the centrifugal device at the rotating speed of 10000-20000 rpm to obtain the thallus active mud.
The invention has the beneficial effects that: according to the thallus culture equipment, the first chamber, the second chamber and the third chamber which are sequentially connected from top to bottom are arranged in the fermentation tank, thallus obtained by fermentation in the first chamber can be enriched by using the second chamber, and the enriched thallus in the second chamber can be fully accumulated and settled by using the third chamber, so that the thallus can be concentrated without additionally arranging a rotary sedimentation tank, a circulating pump, a circulating pipe and other devices, the equipment structure is simplified, and the convenience in operation and control of the equipment and the production efficiency are improved; meanwhile, the stirring paddles of the stirring device are positioned in the first chamber and the second chamber, so that on one hand, the culture medium and the thallus seed liquid added into the first chamber can be stirred to ensure that the thallus seeds fully absorb nutrition to achieve a good fermentation culture effect, and on the other hand, the thallus enriched in the second chamber can be stirred to keep the activity of the thallus; in addition, through set up turbidity detection device in the third chamber, do benefit to the turbidity of liquid in the real-time detection third chamber to be connected with the leakage fluid dram of third chamber lower extreme with centrifugal device's inlet through vertical pipeline, do benefit to in time discharging the material to centrifugal device after satisfying the turbidity requirement, in order to obtain the active mud of thallus with the quick centrifugal separation of material. According to the thallus culture method, thallus is produced by adopting the thallus culture equipment, and culture solution with the same volume is supplemented for continuous culture after materials are discharged, so that the culture and separation of the thallus can be carried out simultaneously, the fermentation time is prolonged, the utilization rate of a substrate is further improved, the product yield is improved, the online separation of the thallus can be realized, the timely separation of the thallus is facilitated, and the good activity is kept.
Drawings
FIG. 1 is a schematic diagram showing an embodiment of an apparatus for culturing microorganisms according to the present invention;
labeled as: the device comprises a first chamber 1, a second chamber 2, a third chamber 3, a stirring device 4, a turbidity detection device 5, a liquid discharge port 6, a vertical pipeline 7, a valve 8 and a centrifugal device 9.
Detailed Description
The invention is further illustrated by the following figures and examples.
As shown in FIG. 1, the apparatus for culturing the bacterial cells comprises a fermentation tank, a stirring device 4, a centrifugal device 9, and a turbidity detecting device 5; the fermentation tank is provided with a first chamber 1, a second chamber 2 and a third chamber 3 which are sequentially connected from top to bottom, and the lower end of the third chamber 3 is provided with a liquid outlet 6; the stirring device 4 is arranged on the fermentation tank, and stirring paddles of the stirring device are positioned in the first chamber 1 and the second chamber 2; the turbidity detection device 5 is arranged in the third chamber 3, and a liquid inlet of the centrifugal device 9 is connected with a liquid outlet 6 through a vertical pipeline 7.
According to the thallus culture equipment, the first chamber 1, the second chamber 2 and the third chamber 3 are arranged in the fermentation tank and are sequentially connected from top to bottom, thallus can be cultured by fermentation through the first chamber 1, thallus can be enriched through the second chamber 2, thallus can be fully accumulated and settled through the third chamber 3, and the volume of a culture solution brought out when materials are discharged is reduced, so that thallus can be concentrated without additionally arranging devices such as a rotary sedimentation tank, a circulating pump, a circulating pipe and the like, the equipment structure is simplified, and the convenience in equipment control and the production efficiency are improved; meanwhile, the stirring paddles of the stirring device 4 are arranged in the first chamber 1 and the second chamber 2, so that on one hand, the culture medium and the thallus seed liquid added into the first chamber 1 can be stirred to enable thallus seeds to fully absorb nutrition to achieve a good fermentation culture effect, and on the other hand, the thallus enriched in the second chamber 2 can be stirred to keep the activity of the thallus; in addition, through set up turbidity detection device 5 in third chamber 3, do benefit to the turbidity of liquid in the real-time detection third chamber 3 to be connected with the leakage fluid dram 6 of 3 lower extremes of third chamber with centrifugal device 9's inlet through vertical pipeline 7, do benefit to in time discharging the material to centrifugal device 9 after satisfying the turbidity requirement, obtain the active mud of thallus with the quick centrifugal separation of material, do benefit to the active maintenance of thallus.
The fermentation tank is a main component of the thallus culture equipment, the first chamber 1 is a main part of thallus culture of the fermentation tank, and the first chamber 1 generally needs to ensure a large enough volume, preferably, the volume of the first chamber 1 accounts for 65-75% of the total volume of the fermentation tank; the second chamber 2 of the fermentation tank is mainly used as a settling chamber of the thallus, the second chamber 2 needs to be smaller than the first chamber 1 to achieve the purpose of enriching the thallus, meanwhile, the volume of the second chamber 2 needs to be capable of accommodating a stirring paddle extending into the second chamber to facilitate stirring to keep the activity of the thallus, and preferably, the volume of the second chamber 2 accounts for 20-30% of the total volume of the fermentation tank; the fermentation tank is provided with a third chamber 3 mainly used for enabling thalli to be fully gathered and settled, the volume of the third chamber 3 is small enough to reduce the volume of culture solution brought out when the thalli are discharged, and preferably, the volume of the third chamber 3 accounts for 2-5% of the total volume of the fermentation tank; a liquid outlet 6 formed at the lower end of the third chamber 3 is mainly used for discharging materials formed after the thalli are fully gathered and settled, and a control switch for opening and closing the liquid outlet 6 is usually arranged at the liquid outlet 6.
In order to facilitate the fermentation culture, enrichment and sedimentation of the thallus, it is preferable that as shown in fig. 1, the first chamber 1 includes a cylindrical main body and an inverted cone-shaped bottom portion disposed at the lower end of the cylindrical main body, the second chamber 2 and the third chamber 3 are both in inverted cone-shaped structures, and the first chamber 1, the second chamber 2 and the third chamber 3 are coaxially disposed. The cylindrical main body of the first chamber 1 is beneficial to stirring and can provide a proper environment for the growth of thalli; the inverted conical bottom of the first chamber 1 is beneficial to the grown thalli to flow towards the second chamber 2; the second chamber 2 with the inverted cone structure is beneficial to the enrichment of thalli and the flow of the enriched thalli towards the third chamber 3; the third chamber 3 with the inverted cone structure is beneficial to the sedimentation of enriched thalli and ensures that only a small amount of culture solution is brought out when materials are discharged.
On the basis, in order to further optimize the structure of the fermentation tank, the diameter of the joint of the first chamber 1 and the second chamber 2 is 40-50% of the diameter of the cylindrical main body of the first chamber 1, so that the thalli in the second chamber 2 cannot easily return to the first chamber 1, and the air in the first chamber 1 can enter the second chamber 2; the diameter of the joint of the second chamber 2 and the third chamber 3 is 30-40% of the diameter of the upper end of the third chamber 3, so that the third chamber 3 can fully enrich and settle the thalli, and excessive culture solution is further prevented from being discharged when materials are discharged; the taper of the third chamber 3 is set to be 0.7: 1-1.2: 1, so that the thalli are fully settled at the bottom of the third chamber 3, the third chamber 3 has enough height, the funnel flow is reduced, the turbidity detection device 5 is favorably installed, and the washing is convenient.
The stirring device 4 is mainly used for stirring the culture medium and the thallus seed liquid added into the first chamber 1 so that the thallus seeds can fully absorb nutrition to achieve a good fermentation culture effect; the stirring device 4 can also stir the bacteria enriched in the second chamber 2 to keep the activity of the bacteria; the stirring device 4 generally comprises a stirring shaft vertically arranged on the fermentation tank and extending into the first chamber 1 and the second chamber 2, a stirring blade arranged on the stirring shaft, and a stirring motor arranged on the fermentation tank and in transmission connection with the stirring shaft; the stirring shaft and the stirring blade jointly form a stirring paddle of the stirring device 4. The stirring motor is used for driving the stirring shaft to rotate, and is preferably a servo motor.
The turbidity detection device 5 is mainly used for detecting the turbidity of the liquid in the third chamber 3 so as to facilitate the judgment of the concentration of the thalli in the liquid by workers; the turbidity detecting means 5 may be various, for example: turbidity detectors, turbidity detection sensors, and the like.
The vertical pipeline 7 is a vertically arranged conveying pipeline and is mainly used for conveying materials discharged by the fermentation tank; for the delivery control, a valve 8 is usually provided on the vertical pipe 7, and the valve 8 may be various, for example: manual valves, solenoid valves, etc.
The centrifugal device 9 is mainly used for separating the thalli in the material, and a solid-liquid centrifuge is usually selected, and a disc centrifuge is preferably selected.
The invention also provides a thallus culture method capable of improving the utilization rate of the substrate and the yield of the product, and the method adopts the thallus culture equipment to produce thallus.
Specifically, the method comprises a fermentation step, a sedimentation step and a separation step;
a fermentation step: firstly, adding a sterilized culture medium into a fermentation tank through a first chamber 1, then adding a thallus seed solution into the fermentation tank through the first chamber 1, and simultaneously starting a stirring device 4 for stirring and fermenting; in this step, the stirring speed of the stirring device 4 is generally based on the ability to suspend the bacterial seeds in the culture medium;the inoculation density of the cell seeds in the cell seed solution is usually 1X 105～5×105CFU/mL；
A settling step: circularly operating according to the sedimentation step, and stopping the stirring device 4 after the fermentation of the thallus seeds reaches the logarithmic growth phase; after 3-8 min, starting the stirring device 4 to stir at the rotating speed of 2-10 rpm for 0.5-2 min, so that part of thalli are settled into the third chamber 3; then, continuing stirring and fermenting according to the stirring mode of the fermentation step; the logarithmic phase is the phase that the specific growth speed reaches the maximum after the microbial fermentation system grows for a certain time when the microbial fermentation system is proliferated;
a separation step: when the turbidity detection device 5 detects that the turbidity of the liquid in the third chamber 3 exceeds a set value, discharging the material to the centrifugal device 9 through the vertical pipeline 7, wherein the volume of the discharged material is 80-150% of the volume of the third chamber 3; after the materials are discharged, the culture solution is supplemented into the first chamber 1, and the volume of the supplemented culture solution is equal to that of the discharged materials; the material discharged into the centrifugal device 9 is subjected to centrifugal separation in the centrifugal device 9 at the rotating speed of 10000-20000 rpm to obtain thallus active mud; in the step, the times of discharging the materials are more than the times of supplementing the culture solution, namely the culture solution is not supplemented after the last time of discharging the materials, and the materials are usually discharged for 2-5 times. The numerical value set in the turbidity detecting apparatus 5 is generally set according to the type of the cultured bacterial cells, and is usually 7000 to 10000 NTU.
According to the thallus culture method, thallus is produced by adopting the thallus culture equipment, and the stirring rotating speed and the settling time are effectively controlled, so that the thallus is favorably cultured and effectively settled into the third chamber 3, the thallus amount in fermentation is effectively reduced, and the utilization rate of a substrate is improved; in addition, the culture solution with the same volume is supplemented after the materials are discharged to continue the culture, the simultaneous culture and separation of the thalli are ensured, the fermentation time is prolonged, the utilization rate of a substrate is further improved, the product yield is improved, the online separation of the thalli can be realized, the timely separation of the thalli is facilitated, and the good activity is kept.
Example 1
The yeast is continuously cultured by utilizing the thallus culture equipment and the thallus culture equipment method provided by the invention;
in this example, the fermenter used had a total volume of about 650L, a cylindrical body of the first chamber 1 having a height of 75cm and a diameter of 80cm, an inverted conical bottom of the first chamber 1 having a height of 20cm and a lower end diameter of 35 cm; the height of the inverted conical second chamber 2 is 100cm, and the diameters of the upper end and the lower end are 80cm and 10cm respectively; the height of the inverted conical third chamber 3 is 53cm, and the diameter of the upper end is 42 cm; the volumes of the first chamber 1, the second chamber 2 and the third chamber 3 respectively account for about 66 percent, 30 percent and 4 percent of the total volume of the fermentation tank.
When culturing, 500L yeast malt extract culture medium is pumped into a fermentation tank, and sterilized, and then the seed solution of Zygosaccharomyces rouxii is inoculated into the fermentation tank with an inoculation density of 105CFU/mL, culturing at 30 ℃ after inoculation, and simultaneously starting a stirring device 4 to stir and ferment at the rotating speed of 200 rpm;
after the culture reaches the logarithmic phase after 24 hours, stopping stirring, and standing the fermentation liquor for 5 min; then, starting the stirring device 4 to stir at the rotating speed of 5rpm for 1min, so that part of thalli is settled into the third chamber 3; then, regulating and controlling the stirring device 4 to 200rpm, and continuing stirring and fermenting; continuing culturing for 5h, then reaching logarithmic phase again, stopping stirring, and standing the fermentation liquor for 5 min; then, starting the stirring device 4 to stir at the rotating speed of 5rpm for 1min, so that part of thalli are settled into the third chamber 3, and performing reciprocating and circulating operation in the way;
when the turbidity detection device 5 detects that the turbidity of the liquid in the third chamber 3 reaches 10000NTU, opening the valve 8 to discharge the material to the centrifugal device 9 through the vertical pipeline 7, wherein the volume of the discharged material is about 30L; after discharging the materials, supplementing a glucose solution with the mass concentration of 3% into the first chamber 1 through a feed inlet of the fermentation tank, wherein the volume of the glucose solution supplemented is equal to that of the discharged materials; the material discharged into the centrifugal device 9 is centrifugally separated at the rotating speed of 10000rpm of the centrifugal device 9 to obtain the yeast active bacterial sludge.
And continuously culturing by continuing the culture operation until all materials are discharged after the glucose solution is supplemented for 2 times, and finishing the production.
The verification proves that 2-2.5 kg of yeast mud can be obtained from every 100L of culture solution of the zygosaccharomyces rouxii cultured in the embodiment, compared with the zygosaccharomyces rouxii cultured by the traditional method (1.8-2 kg of yeast mud can be obtained from every 100L of culture solution), the yield is obviously improved, and the viable count of the active lyophilized powder prepared by adding the protective agent is not obviously different from that of the active lyophilized powder prepared by the traditional method.
Example 2
By utilizing the thallus culture equipment and the thallus culture equipment method provided by the invention, lactobacillus plantarum is continuously cultured;
in this example, the fermenter used had a total volume of about 2500L, a cylindrical body of the first chamber 1 having a height of 1m and a diameter of 1.36cm, an inverted conical bottom of the first chamber 1 having a height of 30cm and a diameter of 43cm at the lower end; the height of the inverted conical second chamber 2 is 107cm, and the diameters of the upper end and the lower end are 136cm and 36.5cm respectively; the height of the inverted conical third chamber 3 is 57cm, and the diameter of the upper end is 57 cm; the volumes of the first chamber 1, the second chamber 2 and the third chamber 3 respectively account for about 68 percent, 30 percent and 2 percent of the total volume of the fermentation tank.
During culture, the 2000LMRS culture medium is pumped into a fermentation tank, and is sterilized, and then the Lactobacillus plantarum seed solution is inoculated into the fermentation tank, wherein the inoculation density is 105CFU/mL, culturing at 37 ℃ after inoculation, and simultaneously starting the stirring device 4 to stir and ferment at the rotating speed of 100 rpm;
after the culture reaches the logarithmic phase after 24 hours, stopping stirring, and standing the fermentation liquor for 8 min; then, starting the stirring device 4 to stir at the rotating speed of 2rpm for 30s, so that part of the thalli is settled into the third chamber 3; then, regulating the stirring device 4 to 100rpm, and continuing stirring and fermenting; continuing culturing for 5h, then reaching logarithmic phase again, stopping stirring, and standing the fermentation liquor for 8 min; then, starting the stirring device 4 to stir at the rotating speed of 2rpm for 30s, so that part of thalli are settled into the third chamber 3, and performing reciprocating and circulating operation in the way;
when the turbidity detection device 5 detects that the turbidity of the liquid in the third chamber 3 reaches 7000NTU, opening the valve 8 to discharge the materials to the centrifugal device 9 through the vertical pipeline 7, wherein the volume of the discharged materials is about 40L; after discharging the materials, supplementing a glucose solution with the mass concentration of 5% into the first chamber 1 through a feed inlet of the fermentation tank, wherein the volume of the glucose solution supplemented is equal to that of the discharged materials; the material discharged into the centrifugal device 9 is centrifugally separated at the rotating speed of 15000rpm of the centrifugal device 9 to obtain the lactobacillus active bacteria mud.
And continuously culturing by continuing the culture operation until all materials are discharged after the glucose solution is supplemented for 3 times, and finishing the production.
According to verification, 1.2-1.8 kg of lactobacillus bacterial sludge can be obtained per 100L of culture solution of the lactobacillus plantarum cultured in the embodiment, compared with lactobacillus plantarum cultured by a traditional method (1-1.5 kg of lactobacillus bacterial sludge can be obtained per 100L of culture solution), the yield is remarkably improved, and the viable count of the lactobacillus plantarum cultured by adding the protective agent to prepare the active lyophilized bacterial powder is not remarkably different from that of the lactobacillus plantarum cultured by the traditional method.
Claims (10)
1. Thallus culture equipment includes fermentation cylinder, agitating unit (4) and centrifugal device (9), its characterized in that: also comprises a turbidity detection device (5); the fermentation tank is provided with a first chamber (1), a second chamber (2) and a third chamber (3) which are sequentially connected from top to bottom, and the lower end of the third chamber (3) is provided with a liquid outlet (6); the stirring device (4) is arranged on the fermentation tank, and stirring paddles of the stirring device are positioned in the first chamber (1) and the second chamber (2); the turbidity detection device (5) is arranged in the third chamber (3), and a liquid inlet of the centrifugal device (9) is connected with a liquid outlet (6) through a vertical pipeline (7).
2. The apparatus for culturing bacterial cells according to claim 1, wherein: the stirring device (4) comprises a stirring shaft which is vertically arranged on the fermentation tank and extends into the first chamber (1) and the second chamber (2), a stirring blade which is arranged on the stirring shaft, and a stirring motor which is arranged on the fermentation tank and is in transmission connection with the stirring shaft; the stirring shaft and the stirring blade jointly form a stirring blade of the stirring device (4).
3. The apparatus for culturing bacterial cells according to claim 1, wherein: and a valve (8) is arranged on the vertical pipeline (7).
4. The apparatus for culturing bacterial cells according to claim 1, wherein: the centrifugal device (9) is a disc centrifuge.
5. The apparatus for culturing bacterial cells according to any one of claims 1 to 4, wherein: the volume of the first chamber (1) is 65-75% of the total volume of the fermentation tank, the volume of the second chamber (2) is 20-30% of the total volume of the fermentation tank, and the volume of the third chamber (3) is 2-5% of the total volume of the fermentation tank.
6. The apparatus for culturing bacterial cells according to claim 5, wherein: the first chamber (1) comprises a cylindrical main body and an inverted cone-shaped bottom arranged at the lower end of the cylindrical main body, the second chamber (2) and the third chamber (3) are in inverted cone-shaped structures, and the first chamber (1), the second chamber (2) and the third chamber (3) are coaxially arranged.
7. The apparatus for culturing bacterial cells according to claim 6, wherein: the diameter of the joint of the first chamber (1) and the second chamber (2) is 40-50% of the diameter of the cylindrical main body of the first chamber (1), and the diameter of the joint of the second chamber (2) and the third chamber (3) is 30-40% of the diameter of the upper end of the third chamber (3).
8. The apparatus for culturing bacterial cells according to claim 6 or 7, wherein: the taper of the third chamber (3) is 0.7: 1-1.2: 1.
9. A method for culturing a bacterial cell, comprising: a cell culture apparatus according to any one of claims 1 to 8, wherein the cell is produced.
10. The method for culturing bacterial cells according to claim 9, wherein: comprises a fermentation step, a sedimentation step and a separation step;
a fermentation step: adding the sterilized culture medium into a fermentation tank through a first chamber (1), adding the thallus seed liquid into the fermentation tank through the first chamber (1), and starting a stirring device (4) to stir and ferment;
a settling step: when the fermentation of the thallus seeds reaches the logarithmic growth phase, the stirring device (4) is stopped; after 3-8 min, starting the stirring device (4) to stir at the rotating speed of 2-10 rpm for 0.5-2 min, so that part of thalli is settled into the third chamber (3); then, continuing stirring and fermenting according to the stirring mode of the fermentation step;
a separation step: circularly operating according to the sedimentation step, when the turbidity detection device (5) detects that the turbidity of the liquid in the third chamber (3) exceeds a set value, discharging the material to the centrifugal device (9) through the vertical pipeline (7), wherein the volume of the discharged material is 80-150% of the volume of the third chamber (3); after the materials are discharged, the culture solution is supplemented into the first chamber (1), and the volume of the supplemented culture solution is equal to that of the discharged materials; and discharging the material into a centrifugal device (9), and performing centrifugal separation on the material in the centrifugal device (9) at the rotating speed of 10000-20000 rpm to obtain the thallus active mud.
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