JPS5817586B2 - Tadanrenzokunoushiyukuhatsukousouchi - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a fermentation apparatus characterized by culturing microorganisms aerobically and at the same time continuously concentrating the culture solution to continuously produce a target product at a high concentration.

The inventor has already filed a patent application for the basic patent of the present invention in 1972-
57161 has been submitted.

The gist is that in the fermentation method of culturing microorganisms aerobically,
The aim was to perform a concentration operation in the latter half of the fermentation at the same time as the fermentation to accumulate the target product at a high concentration.

The advantage is that in conventional fermentation methods, the concentration of the target product is low at the end of fermentation, and it is vulnerable to fluctuations in temperature, pressure, and pH, so the cost of separating and recovering the target product is a large proportion of the cost of the fermented product. However, in nature, at the end of fermentation, the target product is taken out as a culture solution with a concentration several to several tens of times higher than in conventional fermentation methods, reducing the cost of separating and recovering the target product and the time required. The aim was to reduce

However, the patent application No. 48-57161 uses the so-called batch fermentation method, and currently most fermented products are produced by this batch fermentation method, but the productivity is lower than that of the continuous method. remains as a drawback.

Also, it is uneconomical to limit the large amount of oxygen-containing gas used for concentration to just one use.

The purpose of the present invention is to improve the drawbacks of the batch fermentation method, and at the same time further improve the above-mentioned Japanese Patent Application No. 48-57161, to continuously extract the desired product in the form of a highly concentrated culture solution. The object of the present invention is to provide a multi-stage continuous concentration fermentation device that is highly productive, labor-saving, and energy-saving.

In conventional batch fermentation methods, fermentation preparation or product separation and recovery steps are concentrated at the start and end of fermentation operation.
Various equipment and labor are not allocated or utilized effectively, and the fermentation equipment itself has shortened operating time due to preparation and product separation and collection work, resulting in a relatively large amount of production per unit time. required equipment capacity.

Therefore, in order to improve the above-mentioned drawbacks, the inventor filed a patent application in 1973.
)-57161 was further developed into a continuous multi-tank type, and a multi-stage continuous fermentation device was completed.

The present invention will be explained in detail below based on the drawings.

The basic unit of the device of the present invention is the fermentation device shown in FIG.
As shown in FIG. 2, five of these are connected in series in the horizontal direction.

In Figure 1, by bringing the culture solution into direct contact with a large amount of oxygen-containing gas and the part 1 where most of the culture solution exists (hereinafter referred to as the culture solution tank), the oxygen in the air is dissolved and at the same time the moisture in the culture solution is removed. The main part is the part that transfers it into the gas (hereinafter referred to as the aeration concentration part) 2, and the liquid transporter 3 transports the culture solution between the two parts.
This is a device designed to circulate water.

In FIG. 1, the culture solution is continuously supplied from the supply pipe 4.

A part of the culture solution in the culture solution tank 1 is extracted from the extraction pipe 5, sent to the top of the apparatus with liquid transport +f93, and dropped from above the aeration concentration section 2.

While the culture solution is falling through the aeration concentration section 2, it is passed through the oxygen-containing gas supply pipe 6.
Oxygen in the oxygen-containing gas supplied to the aeration concentration section 2 is dissolved in the culture solution, and at the same time, water in the culture solution is transferred to the gas.

The culture solution then returns to the culture solution tank 1 again.

A portion of the culture solution is continuously drawn out from the tube 7 and sequentially supplied to the culture solution supply tube 4 of the next stage device.

The oxygen-containing gas enters through the supply pipe 6, takes in water from the culture solution, exits through the discharge pipe 8, and is sequentially supplied to the oxygen-containing gas supply pipe 6 of the preceding stage device.

Figure 2 shows the basic unit device of Figure 1 in the horizontal direction I, n, I.
, IV are connected in series in the order of FIG.

The number of devices connected in series is not limited to four, and may be any number.

The culture solution is supplied to the first fermentation device from the supply pipe 9, and while fermentation is being carried out, in a steady state at equilibrium, the concentrated fermentation solution is extracted from the discharge pipe 10 of the last stage device (①); ,
The culture solution is automatically discharged from the first device and enters the second device by the difference in the level of the culture solution that is generated depending on the total amount of water evaporated in the second and second devices.

The same thing is done in the No. 2 device, and the culture solution is sequentially fermented and evaporated while being sent from the No. 1 device to the No. It will be done.

The oxygen-containing gas is sent by the feeder 11, where it is dedusted and sterilized as necessary, and the temperature and humidity are adjusted to predetermined values by the air conditioner 12.Then, it enters the last stage, the device No. Through air-liquid contact with the culture solution, oxygen is supplied to the culture solution and water is concentrated by evaporation from the culture solution. The contained gas is finally exhausted from the exhaust port 13 of the first device.

That is, the flow of the culture solution and the oxygen-containing gas becomes a series counterflow.

The flow rate of oxygen-containing gas is determined by the amount of water to be evaporated, since it necessarily greatly exceeds the amount of oxygen required for fermentation.

Therefore, even after the introduced oxygen-containing gas is almost saturated with water, if it is brought into direct contact with the culture solution, sufficient oxygen necessary for fermentation will be supplied.

In a culture solution containing volatile substances such as lower alcohols as assimilation substances, a portion of these volatile substances also migrate into the oxygen-containing gas as water evaporates.

In this case, the final discharge port 13 may be equipped with a suitable condenser to collect the water together with the water, adjust the concentration of assimilated substances, etc., and then return it to the culture solution supply pipe 9.

The present device is particularly suitable for non-growth-linked fermentation where there is a time lag between the growth of microorganisms and the production of the desired product.

In other words, by performing the growth of microorganisms and the production of the desired product in separate devices, the conditions within each device can be optimized, resulting in a higher yield of the desired product and improved productivity. can be measured.

In particular, in the case of the device of the present invention, the purpose of each unit device is, for example, the first stage device has the main purpose of growing microorganisms;
The main purpose of the device in stage 2 is to produce the desired product and its secondary purpose is concentration, the device in stage 2 has the main purpose of concentrating the desired product and its secondary purpose is to produce the desired product, and It is possible to optimize the cultivation and concentration operations such that the main purpose is only concentration of substances, and productivity can be improved.

Next, the present invention will be described in more detail with examples.

Example 1 The multi-stage continuous fermentation apparatus shown in FIG. 2 was used to produce amylase by fermentation.

As the gas-liquid contact material for the aeration/concentration section, a large number of metal plates with a number of spherical protrusions attached thereto, of the type used in commercially available general cooling towers, were filled vertically into the apparatus.

The inner diameter of each device is 30 cm, and the height of the part filled with gas-liquid contact material is I and II.

40 cm and 3 for II and IV devices, respectively.
2cm.

22cm and 21cm, the capacity of the culture medium reservoir is 301cm.
It is.

The culture solution flows in the order of devices 1-11-I-IV, and the air flows in the order of devices IV-1-11-I.

50 g/l of soluble starch, Na2HPO4 in the equipment of
Hl 2I (204&/l, KH2PO40.6g
/l.

KCl O,051/A, peptone 19 g#, meat extract 0.5. ! il'/7. MgSO47H200
,5g/11゜CaCA2・2 H2O0,1g/l!
Pour 94ml of culture solution containing pH 7.0 and heat at 121°C1k.
Heat sterilize with gA11L for 10 minutes.

20.51 of tap water for the devices ■, ■, and ■, respectively.
16.31, 12.31, and similarly 1210C,
Sterilize at 1 kg/ci.

After cooling each device to 35°C with GC, 1. OA was inoculated into the device (2), and while the temperature was maintained at 35°C, the culture solution was circulated by a pump at a rate of 351/min, and air was passed at a flow rate of 204/min.

5 hours after the start of culture When the inoculum concentration is 2 mt/ml and the amylase concentration is 12 u/m/ml, the sterilized culture solution with the above composition is
．． It was supplied at a flow rate of 31/hour, while the temperature was 33°C and the humidity was 0.
Sterilized air adjusted to 023 kg H20/kg dry air is supplied to device ① at a flow rate of 155.5 kg/hour, and the flow rate of the culture solution is 1-1 between each device.
．． 92M/hour, n-i is 1.557/hour, I-JV is 0.81! / It was time.

The culture solution circulation rate of each device was 351/min.

After 5 days, the device was in steady state.

1 at this time. The amylase concentrations of each device II, III, and IV are shown in the table below.

For comparison, data obtained when a normal aerated stirring tank of the same capacity is used is also shown.

[Brief explanation of the drawing]

FIG. 1 is an explanatory diagram of the basic unit of the device of the present invention, and the second
The figure is a detailed explanatory diagram of the present invention. 1... Culture solution tank, 2... Ventilation concentration section,
3...Liquid transporter, 4...Culture solution supply pipe, 5...Circulating culture solution extraction pipe, 6...
Oxygen-containing gas supply pipe, 7...Culture solution extraction pipe, 8
......Oxygen-containing gas discharge pipe, 9...Culture solution supply pipe, 10...Concentrated culture solution extraction pipe, 11.
. . . Oxygen-containing gas supply machine, 12 . . . Air conditioner, 13 . . . Moisture-containing oxygen-containing gas discharge pipe.

Claims (1)

[Claims] 1 A device for aerobically cultivating microorganisms, in which two or more fermenters are connected horizontally in series, supplying the culture solution to the first fermenter and supplying oxygen-containing gas to the last fermenter. Then, by making the movement of the culture solution and the movement of the oxygen-containing gas countercurrent and bringing the gas and liquid into direct contact, the oxygen necessary for fermentation is supplied, and at the same time, water is sequentially and continuously moved into the oxygen-containing gas. A multistage concentration fermentation device characterized by continuously taking out a concentrated culture solution from a fermentation device in stages.