WO2013035851A1

WO2013035851A1 - Filling device and filling method

Info

Publication number: WO2013035851A1
Application number: PCT/JP2012/072934
Authority: WO
Inventors: 隆文山口
Original assignee: ダイヤニトリックス株式会社
Priority date: 2011-09-08
Filing date: 2012-09-07
Publication date: 2013-03-14
Also published as: CN103781897A; JP6052178B2; US20140209204A1; CN103781897B; JPWO2013035851A1

Abstract

The present invention relates to a filling method that has a filling step in which a valve (2) at the tip of a vertically movable nozzle (3) of a filling device (10) is disposed within a container (30) and a liquid containing at least either or both of cells of a microorganism and a substance treated thereby transferred from a storage tank (20) is sprayed from the valve (2). In the filling step, either or both of an initial flow rate for filling and a later stage flow rate for filling are less than the maximum flow rate. In addition, the present invention relates to a filling method in which, at the initial filling stage in this filling step, the valve (2) at the tip of the movable nozzle (3) is disposed in the proximity of the bottom surface of the container (30), and thereafter, the movable nozzle (3) is made to rise according to the filling conditions for the liquid in the container (30). According to the present invention, a filling device and filling method that can efficiently fill a container with liquid containing at least either or both of cells of a microorganism or a substance treated thereby can be provided.

Description

Filling apparatus and filling method

The present invention relates to a filling apparatus and a filling method for filling a container with a liquid containing microorganism cells stored in a storage tank and / or a processed product thereof.
This application claims priority based on Japanese Patent Application No. 2011-196036 for which it applied to Japan on September 8, 2011, and uses the content here.

In recent years, a movement to use a biocatalyst has been actively promoted from the viewpoint of effectively utilizing a biochemical reaction having characteristics such as energy saving, resource saving, and low pollution process for production of useful substances. As the biocatalyst, animal cells, plant cells, cell organelles, microbial cells, processed products thereof, and the like containing an enzyme that catalyzes the target reaction are used. Examples of such enzymes include nitrile hydratase having nitrile group hydrating ability, nitrilase having nitrile group hydrolytic ability, or amidase having amide group hydrolytic ability. The use of these enzymes enables inexpensive production of amides or carboxylic acids that are important in the chemical industry.
In recent years, due to the widespread use of biocatalysts in the industrial process field, improvements or proposals have been made not only on the reaction mode of enzyme reactions but also on peripheral technologies such as biocatalyst storage methods and filling methods. For example, in Patent Document 1, as a method for preserving a liquid containing a fungus body or a treated product of a microorganism before being subjected to heat sterilization treatment, for example, a microorganism containing nitrile hydratase, the pH of the liquid is set to 6-8. A method of aeration and agitation while adjusting to the above has been proposed.

Japanese Patent No. 4305332

A liquid containing microbial cells and / or a processed product thereof easily foams. Therefore, there is a problem that foaming occurs when the filling container is filled from the storage tank storing the liquid, and the filling rate per container volume is reduced.
The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a filling apparatus and a filling method capable of efficiently filling a container with a liquid containing microbial cells and / or a processed product thereof. .

The present invention for solving the above problems has the following aspects.
[1] A filling device that fills a container with a liquid containing microbial cells and / or processed products thereof stored in a storage tank,
A filling unit that includes a nozzle that enters the container and discharges the liquid transferred from the storage tank into the container, and the liquid discharge unit of the nozzle is in the filling liquid halfway through the filling of the liquid. apparatus.
[2] A filling device that fills a container with a liquid containing microbial cells and / or processed products thereof stored in a storage tank,
A nozzle for discharging the liquid transferred from the storage tank into the container, a detecting means for detecting a filling state of the liquid in the container, and a flow rate of the liquid discharged from the nozzle A filling apparatus comprising: a flow rate control unit that controls the filling state of the liquid.
[3] A filling device for filling a container with a liquid containing microorganism cells and / or processed products thereof stored in a storage tank,
A movable nozzle capable of moving up and down to discharge the liquid transferred from the storage tank into the container, a detecting means for detecting a filling state of the liquid in the container, and a position of a tip of the movable nozzle, A filling apparatus comprising nozzle position control means for controlling according to a filling state of the liquid in the container.
[4] The filling device according to any one of claims 1 to 3, wherein the storage tank is located above the filling device and is configured to fill the container with the liquid by gravity.
[5] The filling device according to any one of [1] to [4], wherein the cells of the microorganism and / or the processed product thereof have nitrile hydratase activity.
[6] A filling method in which a container is filled with a liquid containing microbial cells stored in a storage tank and / or a processed product thereof,
A nozzle for discharging the liquid transferred from the storage tank into the container, a detecting means for detecting a filling state of the liquid in the container, and a flow rate of the liquid discharged from the nozzle A filling step of disposing the tip of the nozzle of the filling device including a flow rate control means for controlling the filling state of the liquid in the container, and discharging the liquid from the tip;
The filling method, wherein the filling step includes making a flow rate at an initial stage of filling and / or a flow rate at a later stage of filling smaller than a maximum flow rate.
[7] The filling method according to claim 6, wherein the flow rate at the initial stage of the filling step is ½ or less of the maximum flow rate.
[8] The filling method according to claim 6 or 7, wherein the flow rate in the latter stage of the filling step is ½ or less of the maximum flow rate.
[9] A filling method in which a container is filled with a liquid containing microbial cells and / or processed products thereof stored in a storage tank,
A movable nozzle capable of moving up and down to discharge the liquid transferred from the storage tank into the container, a detecting means for detecting a filling state of the liquid in the container, and a position of a tip of the movable nozzle, A filling step of disposing the tip of the movable nozzle of the filling device comprising a nozzle position control means for controlling according to the filling state of the liquid in the container, and discharging the liquid from the tip;
A filling method in which a tip of the movable nozzle is arranged in the vicinity of a bottom surface of the container at an initial stage of filling in the filling step, and then the movable nozzle is raised according to a filling state of the liquid in the container.
[10] The filling method according to any one of [6] to [9], wherein the cells of the microorganism and / or the processed product thereof have nitrile hydratase activity.

That is, the present invention relates to the following.
(1) A filling device for filling a container with a liquid containing at least one of the microorganisms stored in the storage tank and the processed product thereof, and enters the inside of the container and is transferred from the storage tank A filling device comprising a nozzle for discharging the liquid into the container, wherein the liquid discharge portion of the nozzle is in the filling liquid in the middle of the filling of the liquid;
(2) A filling device for filling a container with a liquid containing at least one of the microorganisms stored in the storage tank and a processed product thereof, and discharging the liquid transferred from the storage tank into the container A nozzle for detecting the filling state of the liquid in the container, and a flow rate control means for controlling the flow rate of the liquid discharged from the nozzle according to the filling state of the liquid in the container. A filling device comprising,
(3) A filling device that fills a container with a liquid containing at least one of microorganisms stored in a storage tank and a processed product thereof, and discharges the liquid transferred from the storage tank into the container A movable nozzle capable of moving up and down, a detecting means for detecting the filling state of the liquid in the container, and a nozzle position for controlling the position of the tip of the movable nozzle according to the filling state of the liquid in the container A filling device comprising control means,
(4) The filling device according to any one of (1) to (3), wherein the storage storage tank is provided at an upper portion of the filling device and configured to fill the container with the liquid by gravity.
(5) The filling device according to any one of (1) to (4), wherein at least one of the cells of the microorganism and the processed product thereof has nitrile hydratase activity,
(6) A filling method in which a container is filled with a liquid containing at least one of microorganisms stored in a storage tank and a processed product thereof, and the liquid transferred from the storage tank is discharged into the container. A nozzle for detecting the filling state of the liquid in the container, and a flow rate control means for controlling the flow rate of the liquid discharged from the nozzle according to the filling state of the liquid in the container. The tip of the nozzle of the filling device is provided in the container and has a filling step of discharging the liquid from the tip, and the filling step has at least one of a flow rate at the initial stage of filling and a flow rate at the late stage of filling at a maximum. The filling method, comprising making the flow rate smaller than the flow rate;
(7) The filling method according to (6), wherein the flow rate at the initial stage of the filling step is ½ or less of the maximum flow rate,
(8) The filling method according to (6) or (7), wherein the flow rate in the latter stage of the filling step is ½ or less of the maximum flow rate,
(9) A filling method for filling a container with a liquid containing at least one of microorganisms stored in a storage tank and a processed product thereof, and discharging the liquid transferred from the storage tank into the container A movable nozzle capable of moving up and down, a detecting means for detecting the filling state of the liquid in the container, and a nozzle position for controlling the position of the tip of the movable nozzle according to the filling state of the liquid in the container A tip of the movable nozzle of a filling device comprising a control means is disposed in the container, and the liquid nozzle is discharged from the tip, and the tip of the movable nozzle is placed in the container at the initial filling stage of the filling step. A filling method in which the movable nozzle is raised in accordance with a filling state of the liquid in the container, and
(10) The filling method according to any one of (6) to (9), wherein at least one of the microorganisms and the processed product thereof has nitrile hydratase activity.

According to the present invention, it is possible to provide a filling apparatus and a filling method capable of efficiently filling a container with a liquid containing at least one of microbial cells and processed products thereof.

It is the schematic which shows the structure of the filling apparatus 10 used in Example 1. FIG.

The filling device and the filling method of the present invention are a filling device and a filling method for filling a container with a liquid containing at least one of microorganisms stored in a storage tank and a processed product thereof.
Here, the microbial cell of the microorganism used in the present invention may be a live cell or a dead cell.
Examples of treated cells include crushed cells (live cells or dead cells), extracts from cells, immobilized products obtained by immobilizing cells or extracts on a carrier, and the like. As the extract, an enzyme (crude enzyme or purified enzyme) is preferable.
Examples of the method for immobilizing the cells or the extract include a comprehensive method, a crosslinking method, a carrier binding method, and the like. The inclusion method is a method in which cells or enzymes are encased in a fine lattice of a polymer gel or covered with a semipermeable membrane of a polymer. The crosslinking method is a method in which an enzyme is crosslinked with a reagent having two or more functional groups (polyfunctional crosslinking agent). The carrier binding method is a method of binding an enzyme to a water-insoluble carrier.
Examples of the carrier used for immobilization include glass beads, silica gel, polyurethane, polyacrylamide, polyvinyl alcohol, carrageenan, alginic acid, agar, and gelatin.

As at least one of the microorganisms and the processed product thereof, those having an enzyme activity that can be used as a biocatalyst are preferable. Examples of the enzyme activity include nitrile hydratase activity, nitrilase activity, or amidase activity. Among these, nitrile hydratase activity is preferable.
Nitrile hydratase activity means the activity of acting on the nitrile group of a nitrile compound to convert it into the corresponding amide compound. Examples of the method for measuring the nitrile hydratase activity include a method of measuring a reaction rate per 1 mg of dried cells when reacted at a reaction temperature of 10 ° C. for 10 minutes.
A filling method for filling a container with a liquid containing at least one of microorganisms stored in the storage tank of the present invention and a processed product thereof includes microorganisms having a nitrile hydratase activity of 50 to 500 U / mg, and It is preferable to use at least one of the processed products.
At least one of the microbial cells having nitrile hydratase activity and the treated product thereof include microbial cells containing nitrile hydratase, nitrile hydratase extracted from the microbial cells, the microbial cells or nitrile hydratase. Examples include immobilized immobilization products.

As the microorganism containing nitrile hydratase, any microorganism can be used as long as it has a catalytic activity (nitrile hydratase activity) for converting nitriles to amides. For example, the genus Bacillus, the genus Bacterium, the genus MicrocrOccus, the genus Brevibacterium, the genus Corynebacterium, the genus Nocardia, the genus Pseudomonas (Pseudomonas) Microbacteria, Rhodococcus, Gordona, Vibrio, Nitrosomonas, Streptococcus, Lactobacilto, Lactobacilto Genus, Sa S. genus Saccharomyces, Endomyces, Aspergillus, Penicillium, Mucor, Rhizopus, terrocac, terrochoc Examples include microorganisms belonging to the genus, Fusarium genus, Agrobacterium genus and the like, and Rhodococcus genus is preferable.
Further, a transformant obtained by obtaining a target gene such as nitrile hydratase, nitrilase, or amidase derived from the microorganism and introducing the gene into an arbitrary host by modifying the gene as it is or artificially by a conventional method Can also be used (see Molecular Cloning 2nd Edition, Cold Spring Harbor Laboratory Press, 1989). Examples of such transformants include E. coli MT10770 (FERM P-14756) transformed with nitrile hydratase from the genus Achromobacter (see JP-A-8-266277), Pseudocardia Nitrile hydratase of Escherichia coli MT10822 (FERM BP-5785) (see JP-A-9-275978) or Rhodococcus rhodochrous species transformed with nitrile hydratase of the genus bacterium (JP-A-4-21379) And the like, and the like.
Any one of the microorganisms may be used alone, or two or more thereof may be used in combination.

The liquid containing at least one of the microbial cells and the processed product thereof is a culture solution obtained by culturing the microorganisms in a liquid medium, a suspension of microbial cells separated from the culture solution and optionally washed, or quiescent cells. Examples include a cell disruption solution obtained by crushing a suspension with ultrasonic waves or metal beads at a low temperature.
As the culture conditions (liquid medium composition, pH, culture temperature, etc.), known culture conditions corresponding to the microorganism to be cultured are employed. For example, in the case of microorganisms having nitrile hydratase activity, carbon sources (sugars such as glucose or fructose), nitrogen sources (inorganic nitrogen sources such as ammonium sulfate, ammonium chloride, or ammonium nitrate), organic substances such as yeast extract, peptone, or meat extract Nitrogen source) and optionally in a liquid medium supplemented with inorganic salts, metal salts, vitamins or the like, the cells are cultured at 20 to 40 ° C. and pH 5 to 9. The culture may be shake culture or stirring culture as appropriate.

The liquid containing at least one of the microorganisms obtained as described above and the processed product thereof is usually stored in a storage tank, and then filled in a container for transporting or storing the liquid.
The storage tank is used to store the liquid from the time it is manufactured until it is filled in the container. Industrially, the storage tank is made of metal, and the temperature of the stored liquid is controlled to a temperature suitable for storage (for example, 5 ° C or less). What can be used is preferably used.
The container is not particularly limited as long as it is suitable for filling with a liquid, and heretofore, a container used for transporting or storing a liquid containing at least one of microbial cells and a processed product thereof is known. Available. Generally, a drum or tank made of metal or resin is used. As the drum, one having an internal volume of about 200 L is usually used. A tank having an internal volume of about 1 to 10 m ³ is usually used.
The filling device and the filling method of the present invention are used when filling the container from the storage tank.

The filling device of the present invention includes a nozzle that enters the inside of the container and discharges the liquid transferred from the storage tank into the container, so that the liquid discharge portion of the nozzle is in the filling liquid from the middle of filling the liquid. It is a thing.
It is preferable that the filling device includes a control device that controls the liquid discharge unit of the nozzle to be in the filling liquid halfway through the filling of the liquid.
In the filling apparatus of the present invention, when the nozzle that discharges the liquid transferred from the storage tank into the container enters the inside of the container, it means that the liquid discharging part of the nozzle is inserted from the upper lid to the inner side of the upper lid. .
In addition, it is impossible to make the liquid discharge part of the nozzle in the filling liquid in the middle of filling because there is no filling liquid in the initial stage of filling, but when the filling liquid has accumulated in the container, This means that the nozzle is inserted into the liquid so that the liquid discharge part of the nozzle is in the liquid. That is, it means that the liquid discharge part of the nozzle is located in the filling liquid in the container.
By filling the container with the liquid in such a state, air entrainment at the time of discharging the liquid from the nozzle is suppressed. This suppresses foaming and improves the filling efficiency per container volume.

In addition to the nozzle, the filling device of the present invention includes a detecting means for detecting the filling state of the liquid in the container, and a flow rate control for controlling the flow rate of the liquid discharged from the nozzle according to the filling state of the liquid in the container. Means.
Examples of the filling state in the container include a filling amount (weight of the filled liquid), a filling rate (a ratio of the volume of the filled liquid to the internal volume of the container), or a filling stage.
Examples of the detecting means for detecting the liquid filling state include a means for detecting the filling state using the weight measurement and the filling rate converted by the liquid level sensor, and sending it to the control device as a signal. Specifically, a weight measuring device, a liquid level sensor, a flow rate sensor, or the like can be given.
The said control apparatus is an apparatus which processes the signal sent from the detection means, and sends the processed signal to a flow control means. The signal processed by the control device is sent to the flow rate control means, and the flow rate control means is operated as necessary to control the flow rate of the liquid. The control device is preferably a device controlled by a computer.
The flow rate control is to make it possible to control the flow rate of the liquid according to the filling state of the liquid in the container by the opening degree of the flow rate control valve installed in the liquid flow path such as a pipe or a nozzle. The flow rate control valve can be provided in a pipe connecting the storage tank and the filling nozzle, in the nozzle, or in the discharge portion of the nozzle.
Examples of the flow rate control means include a flow rate control valve, a valve, or a flow rate control of a pump.

In the filling apparatus of the present invention, it is preferable that the nozzle is a movable nozzle that can move up and down. In this case, it is preferable that the filling device further includes nozzle position control means for controlling the position of the tip of the movable nozzle in accordance with the state of filling of the liquid in the container.
The movable nozzle that can move up and down is a nozzle that can move up and down in a direction perpendicular to the bottom of the container.
As described above, the signal sent to the control device by the detection means is processed by the control device. When the nozzle is a movable nozzle that can move up and down, the signal processed by the control device is sent to the nozzle position control means, and the nozzle position control means is operated as necessary to control the position of the tip of the nozzle. . The control device is preferably a device controlled by a computer.
The nozzle position control means may be any method as long as it can control the position of the nozzle tip (height from the bottom surface to the nozzle tip) according to the liquid filling condition in the container. Absent.
Examples of the nozzle position control means include a nozzle lifting / lowering device or a nozzle expansion / contraction device.

In the filling device of the present invention, it is preferable that the storage tank is located above the filling device and is configured to fill the container with the liquid by gravity.
When the storage tank is at the top of the filling device and the container is filled with the liquid by gravity, the liquid level of the liquid containing at least one of the microorganisms stored in the storage tank and the processed product thereof is always At the top of the filling device, that is, higher than the top lid of the container placed in the filling device, the liquid in the storage tank is transferred to the container and filled by gravity and the opening of the flow control valve without using a pump. Say to do.

The filling device according to one aspect of the present invention includes:
A storage tank for storing a liquid containing at least one of microbial cells and processed products thereof;
A container for transporting or storing the liquid;
A nozzle for discharging the liquid transferred from the storage tank into the container;
Detecting means for detecting the filling state of the liquid in the container;
It is preferable to include a flow rate control unit that controls the flow rate of the liquid discharged from the nozzle in accordance with the filling state of the liquid in the container.

Another aspect of the present invention is a filling device.
A storage tank for storing a liquid containing at least one of microbial cells and processed products thereof;
A container for transporting or storing the liquid;
A movable nozzle capable of moving up and down to discharge the liquid transferred from the storage tank into the container;
Detecting means for detecting the filling state of the liquid in the container;
It is preferable to include nozzle position control means for controlling the position of the tip of the movable nozzle according to the filling state of the liquid in the container.

The filling method of the present invention is a method using the filling device, and is a method of filling the liquid into the container so that the liquid discharge part of the nozzle is in the filling liquid from the middle of the liquid filling. . Specifically, the filling method of the present invention includes a filling step in which a tip of a nozzle is disposed in a container and liquid is discharged from the tip.
The filling method of the present invention is preferably carried out using a filling apparatus comprising the nozzle, the detection means, and the flow rate control means described above.

In the filling step, it is preferable that at least one of the initial flow rate and the late flow rate be smaller than the maximum flow rate.
The initial stage of filling indicates a stage where the filling rate of the container is less than 12.5%, and the latter stage of filling indicates a stage where the filling rate is 97.5% or more. The stage between the initial stage of filling and the latter stage of filling is called the middle filling stage, and the flow rate in the middle filling stage is the maximum flow rate.
The flow rate is a volume per unit time of the liquid discharged from the nozzle, and is measured by a flow meter, a weight measuring device, or the like.
When the liquid flow rate is the maximum flow rate in the initial stage of filling, that is, when there is almost no liquid in the container, the discharge linear velocity is high, and foaming is likely to occur due to contact between the liquid and the inner wall of the container or air entrainment. By making the liquid flow rate at the initial stage of filling smaller than the maximum flow rate, foaming of the liquid can be suppressed, and the filling efficiency per container volume can be increased.
In addition, by making the amount in the later stage of filling smaller than the maximum flow rate, the filling amount can be easily adjusted at the end of filling, and the filling efficiency per container volume can be increased.
The initial, middle, or late filling rate setting value is not restricted by the above setting value, but should be optimized according to the bottom area and height of the filling container, and the viscosity and ease of foaming of the filled bacterial solution. Is preferred.

The maximum liquid flow rate (medium flow rate) is preferably set so that the liquid discharge linear velocity at the valve at the tip of the movable nozzle is 0.2 to 3 m / sec, and is 0.2 to 2 m / sec. It is more preferable to set so that.
The discharge linear velocity is a value obtained by dividing the discharge flow rate by the nozzle discharge port cross-sectional area, and is measured by a flow meter, a weight measuring device, or the like.
The initial flow rate should be less than the maximum flow rate, but is preferably 1/2 or less of the maximum flow rate from the viewpoint of excellent effect of improving the charging efficiency. In particular, 1/5 or less of the maximum flow rate is more preferable.
The lower limit is not particularly limited, but considering the time required for filling, 1/100 or more of the maximum flow rate is preferable.
That is, the range of the initial flow rate is preferably 1/100 or more and 1/2 or less, more preferably 1/100 or more and 1/5 or less of the maximum flow rate.
The flow rate at the end of filling should be less than the maximum flow rate, but is preferably 1/2 or less of the maximum flow rate from the viewpoint of excellent foaming suppression effect. In particular, 1/5 or less of the maximum flow rate is more preferable. The lower limit is not particularly limited, but considering the time required for filling, 1/100 or more of the maximum flow rate is preferable.
That is, the range of the flow rate in the later stage of filling is preferably 1/100 or more and 1/2 or less, more preferably 1/100 or more and 1/5 or less of the maximum flow rate.

The filling method of the present invention includes a movable nozzle that can move up and down as a nozzle, and further includes a nozzle position control unit that controls the position of the tip of the movable nozzle in accordance with the state of filling of the liquid in the container. It is preferable that the tip of the movable nozzle is disposed in the vicinity of the bottom surface of the container at the initial stage of filling in the filling step using a filling device, and then the movable nozzle is raised according to the filling state of the liquid in the container.
The vicinity of the bottom surface of the container means a height of the container height of 20% from the bottom surface.
Raising the movable nozzle according to the state of liquid filling means that the movable nozzle is submerged in the liquid so that the liquid discharge part of the movable nozzle is in the liquid filling after the filling liquid is accumulated in the container. It means to be in a state where it is plugged in. By causing the liquid discharge portion of the movable nozzle to be present in the filling liquid, air entrainment during the discharge of the liquid from the movable nozzle is suppressed. This suppresses foaming and improves the filling efficiency per container volume.
When the working nozzle is inserted into the filling liquid, the length from the filling liquid surface to the discharge section is preferably 1 to 50 cm.
Further, by raising the liquid discharge portion of the movable nozzle in accordance with the rise of the liquid level, the volume of the nozzle immersed in the filling liquid can be minimized, leading to an improvement in filling efficiency.
The volume of the nozzle immersed in the filling liquid is preferably 0.00002 to 0.005 m ³ .

As described above, according to the filling apparatus and the filling method of the present invention, foaming at the time of filling is performed by controlling at least one of the flow rate and the position of the tip of the nozzle according to the filling state of the liquid in the container. And the filling efficiency per container volume can be improved.

Hereinafter, the present invention will be specifically described with reference to examples and comparative examples, but the present invention is not limited to these examples within the scope of the gist thereof.

[Example 1]
(1) About Embodiment FIG. 1 is a schematic diagram showing a configuration of a filling apparatus 10 used in this example.
The filling device 10
A pipe 1 having an upstream end connected to a storage tank 20 for storing a liquid L (bacterial cell suspension) containing at least one of the microorganisms and the processed product thereof;
A vertically movable movable nozzle 3 provided at the downstream end of the pipe 1;
A valve 2 provided at the tip of the movable nozzle;
A nozzle lifting and lowering device 4 for moving the movable nozzle 3 up and down;
A weight measuring device 5 for measuring the weight of the container 30;
A control device 6 electrically connected to the valve 2, the nozzle lifting device 4, the weight measuring device 5;
Is provided.
A container 30 is placed on the weight measuring device 5, and a storage tank 20 is installed above it.
Specifically, as the valve 2, a nozzle having a lid at the tip of the nozzle and adjusting the flow rate by opening and closing the lid slightly is used. The container 2 can be filled with the liquid L in the storage tank 20 by the valve 2 and gravity.
A followable long nozzle type was used as the movable nozzle 3.
As the nozzle lifting device 4, a hydraulic lifting device was used.
As the weight measuring device 5, a load cell type scale was used.

(2) Preparation method of bacterial cell suspension Rhodococcus rhodochrous J-1 strain (FERM BP-1478) having nitrile hydratase activity was prepared by using glucose 2 mass%, urea 1 mass%, peptone 0.5 Cultured aerobically in a medium (pH 7.0 at 20 ° C.) containing 10% by mass, 0.3% by mass of yeast extract and 0.05% by mass of copal chloride. After completion of the culture, the cultured cells were collected by centrifugation, and washed with 50 mM phosphate buffer (pH 7.0 at 20 ° C.). The buffer solution was added to the washed cells to obtain a cell suspension (10% by mass in terms of dry cells).

(3) Filling the container from the storage tank of the bacterial cell suspension 3000 L of the above-mentioned cell suspension (10% by mass in terms of dry cells) is added to a 9 m ³ volume stainless steel storage tank 20, and the container 30 Filling was started. At the time of filling, the valve 2 is slightly opened and filled at 0.5 L / min when the filling rate is 12.5% or less and when the filling rate is 97.5% or more. Was fully opened and filled at 20 L / min.
At the time of filling, the position of the movable nozzle 3 is adjusted by the control device 6 so that the valve 2 at the tip of the movable nozzle 3 is always immersed in the cell suspension at a certain depth according to the weight measurement value in the weight measuring device 5. Controlled.
As a result, the cell suspension could be filled into the container 30 while suppressing foaming.
In this way, foaming when filling the cell suspension was suppressed, and the volume of the movable nozzle 3 immersed in the filled cell suspension could be minimized. , Filling efficiency improved.

According to the present invention, it is possible to provide a filling apparatus and a filling method capable of efficiently filling a container with a liquid containing at least one of microbial cells and processed products thereof. Therefore, the present invention is useful in the use of biocatalysts in the industrial process field.

1: Piping 2: Valve 3: Movable nozzle 4: Nozzle lifting device 5: Weight measuring device 6: Control device 10: Filling device 20: Storage tank 30: Container L: Liquid

Claims

A filling device for filling a container with a liquid containing at least one of microbial cells stored in a storage tank and a processed product thereof,
A filling unit that includes a nozzle that enters the container and discharges the liquid transferred from the storage tank into the container, and the liquid discharge unit of the nozzle is in the filling liquid halfway through the filling of the liquid. apparatus.
A filling device for filling a container with a liquid containing at least one of microbial cells stored in a storage tank and a processed product thereof,
A nozzle for discharging the liquid transferred from the storage tank into the container, a detecting means for detecting a filling state of the liquid in the container, and a flow rate of the liquid discharged from the nozzle A filling apparatus comprising: a flow rate control unit that controls the filling state of the liquid.
A filling device for filling a container with a liquid containing at least one of microbial cells stored in a storage tank and a processed product thereof,
A movable nozzle capable of moving up and down to discharge the liquid transferred from the storage tank into the container, a detecting means for detecting a filling state of the liquid in the container, and a position of a tip of the movable nozzle, A filling apparatus comprising nozzle position control means for controlling according to a filling state of the liquid in the container.
The filling device according to any one of claims 1 to 3, wherein the storage tank is provided at an upper portion of the filling device and configured to fill the container with the liquid by gravity.
The filling device according to any one of claims 1 to 4, wherein at least one of the microorganisms and the processed product thereof has nitrile hydratase activity.
A filling method for filling a container with a liquid containing at least one of microorganisms stored in a storage tank and a processed product thereof,
A nozzle for discharging the liquid transferred from the storage tank into the container, a detecting means for detecting a filling state of the liquid in the container, and a flow rate of the liquid discharged from the nozzle A filling step of disposing the tip of the nozzle of the filling device including a flow rate control means for controlling the filling state of the liquid in the container, and discharging the liquid from the tip;
The filling method, wherein the filling step includes making at least one of a flow rate at an early stage of filling and a flow rate at a later stage of filling smaller than a maximum flow rate.
The filling method according to claim 6, wherein a flow rate at an initial stage of the filling step is ½ or less of a maximum flow rate.
The filling method according to claim 6 or 7, wherein the flow rate in the latter stage of the filling step is ½ or less of the maximum flow rate.
A filling method for filling a container with a liquid containing at least one of microorganisms stored in a storage tank and a processed product thereof,
A movable nozzle capable of moving up and down to discharge the liquid transferred from the storage tank into the container, a detecting means for detecting a filling state of the liquid in the container, and a position of a tip of the movable nozzle, A filling step of disposing the tip of the movable nozzle of the filling device comprising a nozzle position control means for controlling according to the filling state of the liquid in the container, and discharging the liquid from the tip;
A filling method in which a tip of the movable nozzle is arranged in the vicinity of a bottom surface of the container at an initial stage of filling in the filling step, and then the movable nozzle is raised according to a filling state of the liquid in the container.
The filling method according to any one of claims 6 to 9, wherein at least one of the cells of the microorganism and the processed product thereof has nitrile hydratase activity.