JP3052846U - Large scale culture system for photosynthetic bacteria - Google Patents

Abstract

(57) [Summary] [PROBLEMS] To mass-culture purely photosynthetic bacteria at low cost without using a pressure-resistant container or the like. It is possible to efficiently perform photosynthesis without forming a light projecting window or arranging a light source in a device main body. It can be implemented without specialized equipment managers. SOLUTION: A sealable container made of FRP, stirring means for stirring a liquid medium before sterilization or a mixture of liquid medium after sterilization and photosynthetic bacteria filled in the container, and operation of the stirring means It has a stirring control means for controlling time, a heating means for heating the contents of the container, a heating control means for controlling the operation of the heating means, and a means for venting the contents of the container. The stirring control means continuously stirs the liquid medium before the sterilization, and intermittently stirs the liquid medium and the photosynthetic bacteria after the sterilization. The heating control means controls the heating of the liquid medium before the sterilization treatment to a boiling temperature state and a predetermined temperature state for promoting the germination of spore-forming germs, and the mixture of the liquid culture medium and the photosynthetic bacteria after the sterilization treatment for the growth of photosynthetic bacteria. Control the temperature to a suitable level.

Description

[Detailed description of the invention]

[0001]

[Technical field to which the invention belongs]

 The present invention relates to an apparatus for purely culturing photosynthetic bacteria in large quantities.

[0002]

[Prior art]

 As an apparatus for purely cultivating a large amount of photosynthetic bacteria, there is a pressure-resistant fermentation tank. This fermentation tank contains a liquid medium for about 7 to 8 minutes in a container body of a pressure-resistant container having a double-wall structure, and is provided with stirring means and aeration means, heating means and pressurizing means inside. . Then, live steam is blown together with air from below the tank through a pressurizing means, and the inside of the container body is maintained at 120 to 130 ° C. and 1.5 to 2 atm. The heating and pressurizing state is continued for about 40 to 60 minutes to sterilize the inside of the container body containing the liquid medium. During this time, ventilation and stirring are performed via the stirring means and the ventilation means. After sterilization, cooling liquid is injected between the inner and outer walls of the pressure-resistant container to cool the liquid medium, and the inoculum is injected. The cultivation is started while maintaining the inside of the container body at about 30 ° C. and performing aeration and agitation, and the cultivation is completed in about 4 days to 1 week.

[0003]

[Problems to be solved by the invention]

 Conventional culture equipment requires expensive equipment such as pressure-resistant vessels, boilers, and coolers. As the temperature of the surrounding environment of the culture device rises, adequate ventilation must be provided. In addition, it is necessary to have a specialist operator who handles pressure vessels and boilers stationed. As a result, these devices tend to have high running costs as well as initial investment.

[0004] Further, the pressure-resistant container does not have translucency due to its material (for example, stainless steel SUS304). For this reason, in order to perform or promote photosynthesis by irradiating the culture with light, it is necessary to provide a pressure-resistant light-emitting window (viewing window) on the tank wall or to arrange a light source in the container body.

[0005] An object of the present invention is to provide an apparatus for large-scale pure culture of photosynthetic bacteria, which can stably perform large-scale pure culture of photosynthetic bacteria relatively inexpensively without using a pressure-resistant container or a boiler. It is in.

Another object of the present invention is to provide a pure culture apparatus for photosynthetic bacteria, which allows efficient photosynthesis without forming a light-emitting window or disposing a light source in the apparatus body. is there.

[0007] Still another object of the present invention is to provide a large-scale pure culture apparatus for photosynthetic bacteria, which can be implemented without the need for an expert manager of equipment.

[0008]

[Means for achieving the object]

 The present invention has the following configuration to achieve the above object. That is, the device of the present invention comprises a sealable container formed of fiber reinforced plastic (FRP), and a liquid medium before sterilization filled in this container, or a liquid medium after sterilization, and a photosynthetic bacterium. Stirring means for stirring the mixture of the above, stirring control means for controlling the operation time of the stirring means, heating means for heating the contents of the container, heating control means for controlling the operation of the heating means, and contents of the container And means for ventilating the air. Then, the stirring control means continuously rotates the stirring blade when the container is filled with the liquid medium before the sterilization, and when the container is filled with the liquid medium after the sterilization and the photosynthetic bacteria, the stirring blade. Is controlled to rotate intermittently. When the container is filled with the liquid medium before the sterilization treatment, the heating control means sets the liquid medium to a boiling temperature state so as to intermittently sterilize various bacteria in the liquid medium and a predetermined temperature state to promote germination of spore-forming germs. When the mixture of the liquid medium and the photosynthetic bacterium after the sterilization treatment is filled in the container, the mixture is controlled to a temperature suitable for the growth of the photosynthetic bacterium.

[0009] The photosynthetic bacteria that can be cultured are not particularly limited, and a typical example is Rhodobaccus capsulata. The heating control means controls the heating means such that, for example, the liquid medium is heated at least two and a half cycles with one cycle being a boiling temperature state and a predetermined temperature state for promoting germination of spore-forming germs.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION

 Hereinafter, the present invention will be described in detail based on illustrated embodiments. FIG. 1 is a front view, partially in section, of an apparatus according to an embodiment of the present invention. In the figure, reference numeral 1 denotes a container serving as an incubator, which is integrally formed of FRP (fiber reinforced plastic). This container 1 has a cylindrical shape without a window. Because FRP is easy to process, it can be formed relatively inexpensively, for example, from a container of about 300 liters to a container of about 10 tons. A lid 2 is attached to the upper part of the container so as to be freely opened and closed. The lid 2 is provided with a sealable exhaust port 3 and an inoculation port 4. Although not shown, the exhaust port 3 is provided with a filter material through which steam can pass.

A sheath heater 5 is laid at the bottom of the container. The seeds heater 5 is appropriately ON / OFF controlled by the control circuit 5a in order to heat and boil the liquid medium or culture the photosynthetic bacteria. Further, a blade 6 for ventilation and stirring is provided in the container. The shaft 6a of the stirring blade 6 is connected to a motor 7 on the lid. The motor 7 is timely driven by the control circuit 7a, and rotates the stirring blade 6 via the shaft 6a to stir the contents of the container. The body of the container 1 is covered by a jacket 8 made of FRP for forcibly cooling the contents. The cooling jacket 8 has a cooling water inlet 8a at a lower part and a cooling water outlet 8b at an upper part.

[0012] Specifically, when the container is filled with the liquid medium before sterilization, the heater control circuit 5a controls the temperature of the liquid medium at the boiling temperature and the spore-forming bacteria so as to intermittently sterilize the various bacteria in the liquid medium. When the mixture is filled with a mixture of the liquid medium after sterilization and the photosynthetic bacteria, the mixture is controlled to a temperature suitable for the growth of the photosynthetic bacteria.

The composition of the liquid medium is appropriately selected according to the type of photosynthetic bacteria to be cultured, and the liquid medium is injected into the container 1 as an incubator for about 8 to 9 minutes. Intermittent sterilization is performed by repeated heating and boiling. For example, the liquid medium is heated to 100 ° C. by the sheath heater 5 and boiled for about 3 hours. Since the steam accompanying the boiling is discharged out of the vessel through the exhaust port 3, the internal pressure of the container does not increase. After heating and boiling for a predetermined time, the heater 5 is once turned off. At the same time, water flows in from the inlet 8a of the jacket 8 and circulates through the jacket to discharge the absorbed heat from the outlet 8b. Thereby, the liquid medium in the container is rapidly cooled. The liquid medium rapidly cooled to about 30 ° C. is kept at a constant temperature overnight while maintaining the temperature (about 30 ° C.).

After the incubation at the constant temperature, the heater control circuit 5a controls the operation of the heater so that the boiling, the rapid cooling, and the incubation at the heater are further repeated once, and then the re-boiling and the re-cooling are performed by the heater. I do. By the first boiling, the germinated bacteria contained in the liquid medium are killed. During the first incubation, the spore-form bacteria contained in the liquid medium germinate. The germinated bacteria are similarly killed by the second boil. If bacteria that did not germinate in the first incubation may germinate in the second incubation, they will die in the third boiling. By repeating boiling and leaving twice, the sterilization state of the liquid medium can be obtained as much as possible. However, it is desirable to perform three times of intermittent sterilization just in case, and the heater control circuit 5a controls the sterilization. .

The heater control circuit 5a also controls the temperature in the container after the sterilization as described above. This temperature control controls the temperature at which the photosynthetic bacterium grows in a liquid medium after the sterilization treatment. The amount of the inoculum to be added to the liquid medium after sterilization is preferably in the range of 1 to 20%, particularly about 10% of the liquid medium.

The control circuit 7a of the motor 7 that drives the shaft 6a of the stirring blade 6 continuously controls the motor 7 while the liquid medium in the container is intermittently sterilized, that is, during a series of heating and cooling processing steps. Drive. The stirring blade 6 receiving the driving force of the motor 7 constantly stirs and ventilates the liquid medium. After the inoculation of the inoculum, the control circuit 7a of the motor 7 rotates the stirring blade 6 intermittently. Stirring is performed at a rotation speed of 50 to 100 rpm for about 10 minutes a day. Frequent agitation is preferably avoided because dissolved oxygen in the contents of the container (mixture of photosynthetic bacteria and liquid medium) tends to cause growth of bacteria other than photosynthetic bacteria.

The culturing days are about 7 to 10 days. During this culture period, the photosynthetic bacteria in the container contents grow preferentially because the liquid medium is sterilized as described above. Furthermore, since the container is molded by FRP, unless the pigment is selected to have a color with poor translucency, light is applied to the contents of the container, making photosynthesis by the photosynthetic bacteria more aggressive and efficient. Will be.

Experimental Example Hereinafter, a specific experimental example in which a photosynthetic bacterium (Rhodobacter capsulata) was cultured using the device of the present invention will be described. In a 10 t culture tank, sodium propionate 0.1% and monopotassium phosphate 0.1% were added. 5%, dipotassium phosphate 0.06%, ammonium sulfate 0.1%, magnesium sulfate heptahydrate 0.02%, sodium chloride 0.02%, calcium chloride dihydrate 0.05%, Bacto yeast extract (Difco) 0.01%, vitamin solution (solution of 50 mg thiamine hydrochloride, 50 mg niacin, 30 mg paraaminobenzoic acid, 10 mg pyridoxyhydrochloride, and 5 mg pithione in 100 ml distilled water) 1 ml / liter, trace elements Solution (1000 mg of EDTA-2Na, FeCl _Two 6H_TwoO 2000mg, ZnCl_Two 100 mg, MnCl_Two・ 4H_TwoO 100 mg, H_ThreeBO_Three 100 mg, CoCl_Two・ 2H_TwoO 100 mg, Na_TwoMo O_Four・ 2H_TwoO 20mg, CuCl_Two・ 2H_TwoO 10mg, NiCl_Two・ 6H_TwoO 10 mg and Na_TwoSeO_Three 9000 liters of a medium containing 1 ml / liter of 5 mg dissolved in 1 liter of distilled water was added, and intermittent sterilization was repeated three times at 100 ° C. for 3 hours. After cooling the medium to 30 ° C., 900 liters of an inoculum having a concentration of 3 g of Rhodopbacter capsulata cultivated in another culture tank and having a wet weight / liter concentration was aseptically inoculated. After stirring at 100 rpm for 10 minutes, the stirring was stopped and culture was performed. Agitation was performed for 10 minutes each day, and culturing was performed for 10 days. As a result, cells with a wet weight of 34 kg were obtained.

[0019]

[Effect of the invention]

 According to the present invention, in a FRP container, the heating control means and the stirring control means perform intermittent sterilization by heating and boiling the liquid medium and standing at a constant temperature, and prepare conditions necessary for the growth of photosynthetic bacteria. Therefore, large-scale pure culture of photosynthetic bacteria can be performed at low cost and stably without using equipment such as a pressure-resistant container and a boiler.

Further, according to the present invention, since a container made of FRP having a light-transmitting property is used for the container, the photosynthesis of the photosynthetic bacteria in the container can be efficiently performed without providing a light transmitting window or a light source in the container. Can be done.

Furthermore, according to the present invention, a large amount of photosynthetic bacteria can be easily and automatically cultured in a large scale without the need for an expert administrator for equipment such as a pressure vessel and a boiler.

[Brief description of the drawings]

FIG. 1 is a front view showing an apparatus according to an embodiment of the present invention in a partial cross section.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 ... Container 3 ... Discharge port 4 ... Inoculation port 5 ... Heater 6 ... Stirring blade 8 ... ·Jacket

Claims (3)

[Utility model registration claims] 1. A sealable container molded of fiber reinforced plastic (FRP), and a liquid medium before sterilization filled in this container is stirred or a mixture of the liquid medium after sterilization and photosynthetic bacteria is stirred. Stirring means for controlling the operation time of the stirring means; heating means for heating the contents of the container; heating control means for controlling the operation of the heating means; and means for venting the contents of the container. The stirring control means is configured to continuously rotate the stirring blade when the container is filled with the liquid medium before the sterilization, and to stir when the container is filled with the liquid medium after the sterilization and the photosynthetic bacteria. When the container is filled with the liquid medium before the sterilization treatment, the heating control means controls the liquid medium to a boiling temperature so as to intermittently sterilize various bacteria in the liquid medium. Heating to a predetermined temperature state that promotes the germination of sporulated germs and the sporulated bacteria, and when the mixture of the liquid medium and the photosynthetic bacteria after sterilization is filled in the container, the mixture is brought to a temperature suitable for the growth of the photosynthetic bacteria. A mass cultivation device for photosynthetic bacteria. 2. The apparatus according to claim 1, wherein said photosynthetic bacterium is Rhodobacta capsulata. 3. The heating control means controls the heating means such that the liquid medium is heated for at least two and a half cycles with one cycle comprising a boiling temperature state and a predetermined temperature state for promoting germination of spore-forming germs. The device according to claim 1.