JPS63164881A - Biosynthetic method and apparatus - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION [Object of the Invention] (Industrial Application Field) The present invention relates to a biosynthetic method and a biosynthetic apparatus used in this method.

BACKGROUND OF THE INVENTION Biosynthetic routes involving biomass cultivation to obtain commercially valuable products have been used for a considerable period of time. Many examples include the brewing industry, such as yeast fermentation, and the pharmaceutical industry, such as penicillins and cephalosporins (c
ephalosporin) found in the production of antibiotics.

Improvements are always needed in implementing such biosynthesis. (Thus, for example, wa fermentation reactions are usually carried out in large-scale fermenters, where it is difficult to control the reaction conditions, e.g. the degree of agitation required for aS yield. It is also difficult to clean.

Other problems encountered include the growth of undesirable microscopic species such as bacteria, amoebas and rotifers within the biomass culture.

The present invention seeks to provide an improved biosynthetic method that can be carried out on a substantially commercial scale using organisms that grow well in the absence or partial absence of light, and an apparatus for use therein. It is something.

[Structure of the Invention] (Means for Solving the Problems) The biosynthesis method of the first invention of the present invention is a biosynthesis method consisting of living plants that can grow in the absence or partial absence of light. flowing the mixture along with nutrients essential for the growth of the plant through a substantially opaque tube wrapped around an upright core structure, and recovering one or more synthetic products from the mixture. .

The substantially opaque tube preferably forms part of a closed circuit in which the synthesis mixture is continuously flowed, while synthesis products are continuously withdrawn from the mixture and nutrients and constituent liquids are continuously added. do.

One method of operating such a closed circuit involves withdrawing a volume of liquid from the circuit, filtering the solid biomass product from the liquid, adding a nutrient composition liquid to the filtrate, and, after sterilization, removing the filtrate. Return to open circuit.

In this method, the recovery of biomass product from the liquid and the addition of nutrient liquid to the filtrate are carried out outside the open circuit;
The filtrate is sterilized before being returned to the open circuit. Thus, sterile conditions within the closed circuit are not lost.

After the filtrate has been sterilized and cooled, carbon dioxide can also be introduced into the filtrate before returning it to the open circuit.

The biosynthetic device of the second invention of the present invention comprises an upright core structure, a substantially opaque tube wrapped around the core structure, and a living plant that can grow in the absence or partial absence of light. and means for flowing the synthetic mixture consisting of the U-compound along with the essential nutrients necessary for plant growth through the coiled tubing and for recovering one or more synthetic products from the U compound.

Conveniently, the tube is opaque polyethylene or polyethylene with an opaque coating.

The core structure is substantially cylindrical in form with the tube helically wrapped onto the cylinder.

The tube may be in the form of a single spiral layer on the core;
Or multiple layers may be provided. Alternatively, the tube can be wrapped in a series of spirally wound concentric plate configurations connected in an upright stack.

Preferably, the synthesis mixture is pumped to the top of the core structure and flowed downward through the wrapped tube under turbulent conditions. Alternatively, the synthesis mixture can be pumped through a tube to the head tank with a suitable gas introduced at the base of the tube.

The method can be carried out under aerobic or anaerobic conditions. Thus, carbon dioxide or air can be supplied to the tube, or other gases such as oxygen or air/oxygen mixtures can also be employed according to the desired synthesis product. Some biosynthetic reactions proceed under anaerobic conditions. In such cases, introduction of such gas is not necessary.

In one preferred form of the invention, the fact that some biosynthetic reactions proceed aerobically, while others proceed aerobically, is incorporated into two or more biosynthetic reactions as described above. - and can be used by continuous operation. The first reactor (or bank of reactors) is used to perform an anaerobic reaction that generates a gas such as carbon dioxide, which gas is released after separation of the initial product biomass. , used in other actors for aerobic reactions that utilize this gas.

According to a particularly preferred form of the invention, ammonia gas is used as the nitrogen source or one of the nitrogen sources. The controlled use of ammonia injection has been found to be advantageous in minimizing the growth of undesirable microscopic species such as bacteria, amoebae and rotifers.

It is believed that the presence of ammonium salts and ammonium ions shuts down such growth and acts as a nutrient source for force value object growth.

Nutrients for the synthesis are provided at least in part by wastewaters such as sugar mill or oil refinery effluents or other carbohydrate effluents with high biological oxygen demand. Biomass is thus produced as a valuable by-product of the wastewater treatment process, as such wastewater is purified during the process.

A particularly preferred tubing material is Borieburene, especially low density polyethylene, which has the valuable advantages of low cost and also resistance to attack by biomass media. The tube is suitably black or blackish opaque, or has an opaque backing, or is coated with paint to provide such a finish. Metal tubes can also be used. If desired, the opaque tube can also be exposed to sunlight to absorb the heat required for the reaction. In this case, the tube can also be provided with a protective coating to prevent deterioration due to sunlight. If desired, the tubes can also be reinforced or coated with resin for reinforcement purposes, for example for pressure reactions.

(Example) The present decoy will be described by way of an example with reference to the accompanying drawings.

The biosynthesis shown in FIG. 1 consists of a core structure 2 (indicated by dotted lines) that is upright and substantially cylindrical.

The core structure has a continuous external surface and can be formed, for example, from a hollow concrete section. According to the size of the reactor, the concrete part is inspected C1
and can have internal passage stairs that allow maintenance personnel to enter the interior and reach equipment located at the top of the core structure. Alternatively, the core structure can be an openwork structure, for example the trademark
exion) J, or the core can be a cylindrical metal network.

On top of the core structure 2 is a horizontally spirally wound tube 6 of substantially opaque material. The preferred material is polyethylene, preferably low density polyethylene; such tubing is low cost, can be easily extruded, and can be wrapped in long lengths. Polyethylene also has the advantage of good corrosion resistance, and is resistant to the chemical conditions of reaction mixtures to a much wider range than materials such as stainless steel, which are conventionally used in bulk fermentation processes. . However, other plastic materials,
For example, opaque plastic polyvinyl chloride can be used, or rubber can also be used. Materials such as darkened glass may also be used if they withstand the conditions of use. A peg (not shown) can protrude from the core structure to support the tube and prevent the wrapping tube from slipping off.

If desired for reinforcement reasons, the tube can also have a reinforced outer coating, for example of resin reinforced with glass fibers or other fibers. This is particularly the case when it is desirable to carry out biosynthesis under considerable pressure.

The core structure can be three-dimensional, for example an existing tank arranged with tube wraps. The tank can be of any desired diameter, for example from 2 to 5 m.

The lower end of the core structure 2 is buried underground, and the lower end 10 of the tube 6 passes underground to a control pump arrangement, generally indicated at 12. The pumping device @ 12, which can be equipped with a diaphragm pump or any other suitable type of pump, is used to pump the tubes 6, the central tube 16 and the pumps added to the stream of the synthesis mixture, such as carbon dioxide and/or air, ammonia, ammonium salts, It is connected to a supply line 14 of nutrients and nitrogen, such as provided by urea, synthetic nutrients, etc. The feed line is conveniently computer controlled. The outlet pipe 15 conveys the synthesis mixture from the pumping device up a central pipe 16 arranged extending upwardly through the hollow center of the core structure 2 to a head tank 18 . The head tank may have a suitable wash trough (not shown) to allow the product stream to be collected in line 20.

For example, the more concentrated product rising towards the top of the mixture in the wash trough can be recovered by means of a weir device. Alternatively, other separation means, such as a hydrocyclone, can be placed in the head tank 18. Although line 20 is shown emerging from the side of head tank 18, it could equally well extend downward from the center of core structure 2 so that product could be collected at the base of the core structure. In good condition. The head tank can also have a removal system to remove generated gases.

Alternatively, the composite mixture can be pumped from the coil pile tube with a suitable gas introduced at the base of the coil pile tube to the head tank 18, and from the tank 18 to the return line 1 to the pump.
It will be appreciated that 6 may have flow meters and/or heat exchangers or such devices incorporated therein if necessary.

The dimensions of the biosynthesis equipment vary according to the biomass production carried out.

Examples of the use of biosynthetic equipment include the fermentation of WI mothers and the cultivation of molds to yield, for example, cef-10sporin or penicillin antibiotics.

Although it will be appreciated that the tube diameter may vary, the tube diameter should be kept small enough to encourage turbulence as the material passes through the coil. Turbulent flow helps prevent undesired fouling inside the pipes, thus increasing the working life of the plant. Preferably the turbulence is of such a degree as to give a Reynolds number of at least 2000.

Of course, the pumping device could be located above ground, but the illustrated biosynthesis device has the pumping device 12 buried underground to maximize the space between each bioreactor. It will be appreciated that in the embodiment shown in FIG. 2, embedding the pumping equipment provides adequate space to allow for facility passageways between biosynthesis equipment and product collection and transportation means.

Such equipment includes equipment for dealing with extreme weather conditions, such as heat exchangers included in the circuit;
And, for example, in cold-night conditions with the possibility of frost, a removable insulating covering placed on top of the biosynthesis apparatus can be installed when necessary. Similarly, under very hot conditions, coolant can be sprayed onto the actor from an external device. Alternatively, the biosynthesis device itself directs the dispersion of coolant into a tube 6, such as the provision of a watering ring, which is fed to the top of the core structure 2 by a feed tube passing above the hollow core structure, when desired. It can also include equipment for Of course, the biosynthetic apparatus does not need to operate in an external environment.

In the shown and actor modifications (not shown), the core structure 2 is provided with means for recovering the product via a central circulating wash trough leading to a tube extending to the bottom of the core structure. and is mounted on a rotating platform. Such an arrangement has the advantage of using rotational speed to create flow characteristics within the tube.

Another method of biomass production suitable for large scale use uses the plant illustrated in FIG.

The plant consists of a pair of substantially opaque coiled tubes 21 and 22 arranged in parallel and each mounted on a suitable core structure (not shown). The coil is, for example, a plastic tube with a length of 500 m and a diameter of 30 III. Although two coils are shown, more coils can be used. The synthetic mixture is fed from a single line 25 via lines 23 and 24 to coils 21 and 22 and to a flow meter 26.
via a pumping device 27 (preferably a diaphragm pump) with recycled reactant pumped. line 25
Before reaching , the recycled reactant passes through a heat exchange unit 28 with an immersion heater 29 . Unit 28 serves to supply heat if necessary in cold weather or to remove heat in hot ambient conditions. Carbon dioxide and air are also supplied in appropriate streams on lines 30J and 31, respectively.
d meter (not shown) to line 25.

At the top of coils 21 and 22, the flowing synthetic mixture passes through return pipe 35 where the mixture passes through bag filter 3.
6 (where the product biomass is collected and periodically withdrawn), an overflow collector device 3 with downward flow to
They come together in a line 32 for passage to a head tank 33 with 4. This bag is a mesh that retains large mature products while allowing smaller growth products to pass through. The bag filter 36 is placed in a filter unit 37 that is supplied with constituent water by line 38 and nutrients by line 39 from a series of nutrient supplies 40 .

The filtrate in the unit 37, with addition of water J3 and ferns, is returned by line 41 to the head tank 33 under the action of a pump 42 responsive to a water level control valve 43.

Excess carbon dioxide (and/or other gaseous products) is recovered from head tank 33 in line 411. When needed, air is transferred to the head tank 33 via an air purge line 45.
supply to. The overflow collector device 34 circulates the synthesis mixture that does not flow down and returns to the pump device 27 in line 46.

Circulation within the system and product withdrawal are controlled by electrically operated pressure control valves. This valve is involved in operation once the optimal synthesis conditions have been established (as determined by optical density measurements, for example) after commissioning.

The circulation system may incorporate a squeeze collection and sterilization circuit, such as that described further below in connection with FIG.

In the apparatus described above, it is observed that the synthesis mixture is pumped into the coil and the product flows downwards under the force of gravity through the return pipe 35. Straight and relatively long downwards,
Return tube 35 to filter bag 36 provides high velocity to prevent clogging.

When using tubular modules, intermediate pumps and/or
or provide for air or steam injection. This is particularly valuable when the reaction medium has a tendency to become viscous, for example during certain fermentation processes.

Figure 4 shows the pressure drop (and when warm, e.g. 35
An improved reactor embodying an arrangement for overcoming the problem (in the case of tube elongation at °C) is schematically described. Thus,
The coil 50 is arranged on this side of the core IO1. The synthesis mixture is pumped into the coil via a recirculation pump and heat exchanger unit 52, but rather than all being introduced at the bottom of the coil, it is introduced along it by a series of inlet lines 53. The necessary reaction gases are also introduced into each line 53 via a feed main 57 having a valve inlet 58 to each line 53. Similarly, exit login 54
is connected to the head tank 55 from each coil section via the rising main pipe 56.
be led to. It is advantageous to configure the rising main pipe 56 so that it can be moved on the side of the coil to accommodate the increase in pipe size. This main tube can thus be flexible or have a rigid but flexible attachment. The length of the tube in each coil section will vary depending on flow rate, tube internal diameter, tower height, etc., but approximately 8 subunits with a length of approximately 300 II and an internal diameter of 20 am have been found to be suitable. ing.

It will be appreciated that the biosynthetic apparatus can be easily assembled and configured in modular form if desired. Thus, the tubular coil is easily constructed of parts with valves and joints that can optionally remove reaction products and/or supply any additional nutrients needed.

This is particularly useful for rapid reactions such as certain fermentation reactions.

Pumping is often desirable (see above) to provide the necessary power to provide high turbulence in the tube, but in some reactions, for example, the product cells are natural products that require extreme haste. In some cases it is desirable to employ lower flow conditions.

In such cases, it is sufficient to use air and/or other gas supplies as lift to maintain circulation. Compressed gas venturi jets or steam jets can be used if necessary. Steam injection is particularly suitable when certain stones require heat for growth.

The methods and apparatus described above are applicable to a wide range of biomass production methods. If desired, the feed system to the reactor can contain selenium, selenium,
It is also possible to control the introduction of one or more trace elements such as cobalt, copper, zinc, gallium and germanium in small amounts. It will be appreciated that the method allows for the production of single strain pure biomass under controlled conditions that yield a pure and consistent product. The auxiliary treatments employed can increase the concentration of valuable products in the biomass.

It will be appreciated that the nutrients supplied to the biosynthetic apparatus and the operating conditions can be varied considerably.

It has been found that there is value in using ammonia gas as one or only nitrogen source. Controlled injection of ammonia gas promotes the growth of some organisms, but inhibits the growth of undesirable microscopic life such as bacteria, amoebas, and rotifers. This is of considerable commercial importance since the proliferation of such microscopic species has been a major problem in previously proposed biomass production methods. Other gases that inhibit bacterial growth can also be used, such as carbon oxide.

Alternatively, or in addition, the bacterial infiltration problem can be alleviated, at least in part, by using ultraviolet irradiation of the transparent portion of the tube. This can be done, for example, by wrapping an ultraviolet radiation coil around one or more transparent parts of the tube and/or by
This can preferably be achieved by encapsulating the UV radiation tube within a particularly enlarged section of the tube.

The actors described above are suitable for use in both aerobic and anaerobic biomass production processes.

Thus, gases such as carbon dioxide or air can be used for example for Spirulina and Chlorella, while air/oxygen mixtures or oxygen alone can be used for methods such as yeast propagation. Anaerobic methods can also be carried out.

According to a further useful embodiment of the invention, two biosynthetic devices or a train of biosynthetic devices can be connected in series, the first biosynthetic device being used for an anaerobic reaction;
On the other hand, the second glue actor uses CO2 in the cultivation of oxygen-producing algae. Such a system will be explained with reference to FIG.

In the flow diagram of FIG. 5, waste fluid with high biological oxygen demand is passed through line 60 to the first row of actuators 61 as nutrients for carbon dioxide producing biological cultures. Carbon dioxide produced in reactor 61 is recovered in line 62. Liquid from reactor 61 is collected by line 63 at filter unit 64 where solid biomass is collected. The carbon dioxide flow in line 62 is combined with liquid from filter unit 64 in line 65 for passage as nutrients to a second row glue actor 66 for culturing carbon dioxide consuming and oxygen producing organisms. let's do it together. Oxygen is recovered in line 67 for any suitable purpose, while liquid recovered in line 68 is filtered in unit 69 to obtain a solid biomass product. The liquid remaining in line 70 has a significantly reduced biological oxygen demand. Both rows of reactors 61 and 6G consist of opaque tubes in accordance with the present invention to carry out the biological reaction in the absence of light. Of course, additional steps can be added and any desired number of biosynthetic devices can be used in each train.

Any other gas produced in the first glue actor can be used in the next biosynthesis device or for other purposes.

The use of multiple polyactors may also prove useful when one polyactor is used for photosynthetic reactions while another polyactor is used to proceed with biosynthetic reactions in the absence of light.

Providing a method that can recirculate the mixture and operate continuously keeps the consumption of gases and nutrients as low as possible with minimal losses. The product gas can be used in any adjacent chemical plant. The heat produced can be used in heat exchange. High biomass density can be obtained. Even derelict and heterogeneous resources can be used, and the size of the device can be easily expanded to meet increasing demands. lowest 10
Maintaining a temperature of °C requires at most low-grade heat even in harsh winters. Suitable temperature for cooling in summer
Desirable to maintain R temperature. The use of turbulent flow conditions allows for long periods of operation before cleaning is required, so that downtime can be kept to a minimum. In either case, cleaning the tube is a relatively easy operation and can be accomplished by passing a cleaning "glob" through the tube. This is in contrast to the problems faced in cleaning conventional fermentation tanks, where contaminants often have to be scraped off the walls.

It will be appreciated that the system described above lends itself to automatic control and the entire module arrangement can be computer controlled, thereby saving manpower. Another advantage is that parts of such multi-modular systems can be switched, if necessary, from the production of one type of product to another without the need for modification of the main plant. It is.

In FIG. 6, the numeral 101 indicates one type of glue actor as shown in FIG. 1, in which the biomass culture is produced under sterile conditions at a processing temperature, for example 28-30°C. Liquid is continuously pumped from the vessel to the detachment head 102 and, if necessary, returned to the vessel 101 by a recirculation pump 112 with a heat exchanger 113. Detachable head 102
Inside, gases generated during processing are discharged into the atmosphere.

Valve 103 is located in tube 115 leading from IJIR head 102, and periodically this valve
llI! It is opened to allow liquid to flow out from the decapitated head 102. This liquid can be a fixed amount of outflow, e.g. 5-10% of the circulating liquid in a closed system, and the valve is arranged at predetermined intervals, e.g. every hour, to deliver 10-20% of the total liquid from the reactor 101. can be operated on. The liquid flowing along the tube 115 enters the filtration thread 104 and from there a solid biomass product B such as spirulina.
is removed from the liquid filtrate. The filtration thread can be one of any of a number of withdrawal types, ie a rotary vacuum filter, a microfilter, an edge filter or a simple Puchner type or even a filter press. The particular form of filter thread is chosen depending on the particular type of biomass culture being produced. The liquid from the filtration thread is transferred to a mixing tank 1 where nutrients are further added from a source 105.
Avoid passing on 06. This nutrient is in liquid form and is added to the liquid in the tank according to an automatic analyzer that shows the values of carbonates, phosphates, nitrates or other chemicals that are appropriate in the liquid. .

Since sterility within reactor 101 may not be maintained in thread 114 and mixing tank 106, the liquid with added nutrients will be free of all hostile bacteria and other viable substances contained in the filtrate. It passes through a sterilizer 107 where it is killed. Before entering sterilization 2, the filtrate is conveniently passed through an ultrafilter unit in the form of a microfilter 116 in order to remove residual solids of at least 0.2 μm. The sterilization system is usually a steam sterilizer. Any residual biomass material and killed bacteria and other materials can be filtered from the sterilization fluid. The liquid is passed from the sterilizer through cooler 108 to reduce the temperature to about 20°C. If desired, the cooled liquid is then introduced into an absorption unit 109 where carbon dioxide from source 110 is added in an amount suitable to react with the alkali hydroxide present in the liquid to form a solution. carbonate and bicarbonate to the required value.

The amount of carbon dioxide added can be automatically controlled by an analyzer operating with an electrically controlled relay system.

This absorption unit 109 can be pressurized to secure the CO2 solution if desired. Sterile inoculum can be introduced into absorption unit 109 from source 114.

The treated liquid is then injected by the dosing pump 111 into the inlet side of the recirculation pump 112 and returned to the closed system of the treated liquid beam actor 101 and breakaway head 102 . An ultrafiltration unit in the form of a microfilter 117 can further be located between the donor pump 111 and the recirculation pump 112 to remove at least 0.2 μl of solids. A heat exchanger 113 can be located between the recirculation pump 112 and the reactor 101 so that the process temperature is at the required value.

The advantage of the arrangement shown in FIG. 6 is that the primary photophobic system remains sterile and the desired biomass production is produced continuously in a sterile manner. Biomass is removed by a bypass unit, where biomass is removed from a liquid and additional nutrient liquid is added to the liquid. The liquid is sterilized before being returned to the open chain system.

[Effects of the Invention] As explained above, according to the present invention, it is easy to control the reaction conditions and keep the apparatus clean, and the proliferation of undesirable species that can only be seen under a microscope is suppressed. ,
Biosynthesis can be carried out in practice on a substantial commercial scale using organisms that grow well in the absence or partial absence of light.

[Brief explanation of the drawing]

FIG. 1 is a perspective view schematically showing an embodiment of the biosynthesis device according to the present invention, FIG. 2 is a configuration diagram illustrating the arrangement of a large number of such actors, and FIG. Flowchart of the two-row biosynthesis apparatus; Figure 4 is a schematic diagram of another form of glue actor; Figure 5 is a diagram illustrating the process of aerobic and anaerobic stages; Figure 6 is a closure for biomass cultivation. This is a flowchart of the system. 2... Core structure, 6... Pipe, 12... Control pump device, 18... Head tank, 21.22... Coiled tube, 26... Total flow, 27... Pump device, 28.
...Heat exchange unit, 29...Immersion heater, 33...
・Head tank, 34... Overflow collector device, 35...
Return pipe, 36... Bag filter, 37... Filter unit, 40... Nutrient supply device, 42...
Pump, 43... Water level control valve, 50... Coil, 51... Core, 52... Heat exchanger unit, 55
...Head tank, 56... Main rising pipe, 58... Valve inlet, 61.66... Reactor, 64... Filter unit, 69... Unit, 101...
Reactor, 102... Detachment head, 103... Valve, 104... Filter thread, 105.106... Mixing tank, 107... Sterilizer, 108... Cooler, 10
9... Absorption system, 110.114... Source, 111
... Donor pump, 112 ... Recirculation pump, 113
...heat exchanger, 115...tube, 116.117...
・Micro filter,

Claims (12)

[Claims] (1) a substantially opaque synthetic mixture of viable plants capable of growing in the absence or partial absence of light, together with nutrients essential to the growth of the plants, wrapped around an upright core structure; A biosynthetic method comprising flowing through a tube and recovering one or more synthetic products from said mixture. (2) The method according to claim 1, wherein the nutrients essential for growth include carbon dioxide and nitrogen sources. (3) The biosynthesis method according to claim 1 or 2, wherein the nitrogen source is ammonia gas. (4) pumping the synthesis mixture to the top of the core structure and flowing it downward through the tube under turbulent conditions. Biosynthetic methods. (5) A biosynthetic method according to any one of claims 1 to 3, comprising pumping the synthesis mixture upwardly through a tube to the top of the upright core structure. (6) After performing the anaerobic synthesis in a tube in which the gas is generated and separating the biomass product, the mixture is transferred to a second substantially 6. A biosynthesis method according to any one of claims 1 to 5, comprising passing the biomass product through an opaque tube and recovering the biomass product from a second tube. (7) the substantially opaque tube forms part of a closed circuit in which the synthesis mixture is continuously flowed, at the same time synthesis products are continuously recovered from the reaction mixture, and nutrients are continuously added; A biosynthetic method according to claim 1, comprising: (8) withdrawing a quantity of liquid from the closed circuit, filtering solid biomass products from the liquid, adding nutrient constituent liquid to the filtrate, and returning the filtrate to the closed circuit after sterilization; A biosynthetic method according to claim 7. (9) having an upright core structure, a substantially opaque tube wrapped around the core structure, means for flowing a synthetic mixture through the wrapped tube and means for recovering one or more synthesis products from the mixture; A biosynthesis device characterized by: 10. The biosynthetic device of claim 9, wherein the core structure is generally cylindrical and the tube is wound horizontally around the cylindrical core. (11) The biosynthesis device according to claim 10, wherein the tube is wound around the core in multiple layers. (12) The tube is made of polyethylene or other suitable plastic material or rubber, which is opaque or provided with an opaque coating, or rubber. Biosynthesis equipment.