JPS63141576A - Continuous microorganism culturing and collecting apparatus by solid method - Google Patents

Abstract

PURPOSE:To efficiently produce useful metabolite in enhanced physiological and growth activity, by blowing air for removing bacteria together with a culture medium into a moving type continuously foamed immobilizing bed in large quantities in a system for continuously culturing and collecting bacteria in large quantities. CONSTITUTION:Growth beds of a microorganism is assembled on the conveyer chain rotating endlessly by a driving apparatus 6 and continuously pass through a reactor 1 in order and circulated. The reactor 1 is provided with a spray nozzle 2, an inlet 3 of air and a blower 7 for feeding large quantities of air to microorganisms in the growth beds 4. The apparatus is provided with a lower part reactor 5 receiving air passing through the growth beds 4 and cultivation liquids and provided with a discharge hole to an outer storage tank 12 for cultivation liquid, a high pressure spray water-spraying nozzle 25 for recovering microorganisms, a receiver 28, an air-heating equipment 9, a vessel 29 for recovering the organisms or the like.

Description

[Detailed description of the invention] [Industrial application field] The present invention is an aerobic bacterium that enhances physiological and growth activity.

This invention relates to a large-scale continuous culture separation device for facultative anaerobic bacteria, yeast, and fungi. This device can be used in industries that can replace the conventional jar fermenter, such as antibiotic production, enzyme production, and amino acid fermentation.

[Conventional technology]

Conventionally, as a method of supplying a large amount of oxygen to microorganisms, a method has generally been adopted in which air is blown in the form of bubbles into a solution through a porous material using a blower or the like.

[Problem that the invention attempts to solve]

In the conventional method, in the case of a culture solution rich in nutrients, oxygen is dissolved in the solution and then taken up by the microorganisms, so the rate of oxygen dissolution into the culture solution cannot keep up with the oxygen consumption by the microorganisms, and the apparent , the culture medium is always in an anoxic state. In other words, there was a drawback in that the frequency of microorganisms coming into contact with oxygen-containing bubbles was extremely low.

[Means for solving problems]

The present invention uses a device as described below to supply microorganisms with sufficient oxygen, thereby increasing the physiological and growth activity of microorganisms such as aerobic bacteria, bifacultative anaerobic bacteria, yeast, and fungi. Furthermore, it relates to a technique for continuously culturing and sorting.

The microorganism production apparatus (hereinafter referred to as Biopr) according to the present invention will be described below.
(abbreviated as ot-α) will be explained in detail using the drawings.

The figure is a schematic diagram showing one embodiment of the present invention, and in the figure (1)
an upper reaction chamber (4) equipped with a spray nozzle (2) for spraying the culture medium into this interior and an air inlet (3);
is attached to a conveyor chain that is endlessly rotated by a driving device (6) in a microbial growth bed, and is continuously circulated through the reaction chamber. (5) is a lower reaction chamber that receives the air and culture solution that have passed through the proliferation bed (4) and is equipped with an outlet to the culture solution storage tank (12) outside the main body; each of the devices described in (1) to (6) above; In addition to the high-pressure spray water ejection nozzle (25) that collects the grown microorganisms (described later) and the liquid receiving tank (28) directly below it, Biopro
The main body of t-a is constructed. Also, (7) is a blower for continuously supplying a large amount of air to the microorganisms in the growth bed (4), and the air passes through the duct (8) to the air inlet (3) of the upper reaction chamber (1). sent to. (9) is a heat exchanger that heats air to a desired temperature as necessary.

(10) is a filter that prevents dust, germs, etc. in the air from entering the device. The air and culture solution discharged from the lower reaction chamber (5) pass through the discharge duct (11) and enter the culture solution storage tank (12). Here, the air passes through an exhaust duct (13) connected to the culture solution storage tank, removes mist by a mist eliminator (14) in the duct, is sterilized as necessary, and then returns to the outside air. is discharged.

The stock solution, which is a nutrient source necessary for the growth of microorganisms, is continuously sent from the stock solution storage tank to the first culture solution storage tank by the stock solution supply pump (24), where it is mixed with the circulating culture solution, which is a stock solution containing microorganisms. The medium is then sent to the reaction chamber by a culture medium circulation pump (15). The first and second culture solution storage tanks and the third and fourth culture solution storage tanks are each maintained at the same water level by a communication pipe (16), and an overflow pipe attached to the second culture solution storage tank (
17) The culture solution increased by the flow of the stock solution into the first culture solution storage tank flows into the third culture solution storage tank, and the culture solution is refreshed and the liquid level is maintained at a constant level. A part of it is returned to the culture solution storage tank by the culture solution circulation pump (15) and used for stirring inside the storage tank, but most of it is sent to the upper reaction chamber, where it is sprayed from a spray nozzle and supplied to the growth bed. A flow meter (31) is provided during the liquid feeding, and the supply amount is adjusted to be optimal.

The reaction chamber and the outside of the reaction chamber are sealed inside the Bioprot-α main body, but air and culture fluid that have passed through the growth bed and left the reaction chamber are collected at the lowest part of the main body, and are stored outside the main body. The liquid is guided to a storage tank (18), and when the storage amount reaches a certain amount by controlling the liquid level in this storage tank, it is sent to a third culture liquid storage tank by a transfer pump (19) and used again for circulation.

The culture solution storage tank is also equipped with a pH adjustment device to maintain the culture solution at an optimal pH. For this purpose, the pH is measured in each storage tank with a pH meter (20), and the pH is measured in the pH adjustment chemical storage tank (21).
) is sent to each storage tank by each chemical injection pump (22), and the pH is adjusted to an appropriate level. At this time, each storage tank is stirred by a portion of the liquid returned from the culture solution circulation pump (15) to promote mixing of the chemical solution and the culture solution.

The culture solution is supplied through a communication pipe (16) connecting each culture solution storage tank.
The overflow pipe (17) sequentially moves the culture solution from the first storage tank to the fourth storage tank, and in the fourth storage tank, the culture solution discharge pump (23 ) is sent to the next step.

Further, the microorganisms grown in the growth bed are recovered by irradiating high-pressure spray water onto the growth bed. Immediately after the fourth reaction chamber, a nozzle (25
), high-pressure water is sent from the high-pressure spray supply liquid storage tank (26) by a high-pressure spray pump (27), and the microorganisms grown in the growth bed are washed away and collected by the liquid emitted from the nozzle. The microorganisms that flowed out from the growth bed enter the microorganism collection tank (29) outside the main body from the liquid receiving tank (28) directly below, and the collected liquid transfer pump (30)
The liquid is then transferred to a centrifuge, where it is separated into a heavy liquid containing microorganisms and a light liquid (supernatant), and recovered. A part of the light liquid portion is returned to the high-pressure spraying liquid supply tank (26) and used repeatedly.

A continuous porous material (internal bubble diameter φ2) is used as the growth bed for microorganisms.
~5m) is used because it has good air permeability and water permeability, and the material has a large surface area. Microorganisms settle on the growth bed and multiply there. The growth bed is rotated once every 30 minutes to 1 hour using a device as shown in the figure, and at each rotation, the amount of bacteria grown is recovered using high-pressure spray water. The feature of this system is that the grown microorganisms are continuously collected, and the microorganism culture becomes an infinite growth system, which enables constant growth stage, that is, synchronous culture. Synchronized cultivation guarantees constant quality of the product. The microorganisms detached with high-pressure spray water are separated by a centrifuge into a heavy liquid containing microorganisms and a light liquid (supernatant), and the heavy liquid is dried and powdered using a method that does not reduce the activity of useful products. A portion of the liquid portion is returned to the culture medium. The culture solution containing the remaining useful products is collected by leaving the solution or turning it into a dry powder by a method that does not reduce its activity.

〔Example〕

The processing capacity of Bioprot-a is 8.0 from the experimental value.
D.

It has been calculated that a culture solution with an organic matter concentration of 20,000 ppm has 10 times the growth bed volume. For example, B,
O, D, 20,000 ppm starch waste liquid per day
Assuming a factory that emits OO tons, Biopr
The bed volume of ot-α is 10rn', and each part of Bioprot-α is designed based on this bed volume. In this case, the production amount of bacterial dry powder containing amylase is approximately 1.0 to 1.5 tons/day. This corresponds to 20 to 30% of the dry weight of the waste liquid, and n, o, o, of the treated liquid are reduced to below x, oooppm.

〔effect〕

By using the method of the present invention, in which a nutrient solution is sprayed with air without cultivating microorganisms in a culture solution, nutrients can be supplied to the surface of microorganisms without delay, and the bacterial cells are coated with the culture solution. Therefore, the surface of the bacterial cells does not dry out, and oxygen is supplied quickly and sufficiently. Furthermore, since the air is pumped by a pressure fan, the culture solution constantly flows over the surface of the microbial cells, so that metabolites of the microorganisms flow out without being accumulated on the surfaces of the microorganisms. Since this nutrient source is continuously supplied, its metabolites are constantly removed from the system.
It will be sh-out. When more than a certain number of microorganisms settle on a growth bed, the microorganisms are exposed to environmental conditions such as substrate stimulation, sufficient oxygen supply, and lack of nutrient sources necessary for growth, resulting in a reduction in the production of useful products for main nutrients. Production is activated. At the same time, since the growth bed is constantly rotating in a constant direction, the microorganisms are physically stimulated at regular intervals, resulting in synchronized growth of the microorganisms.

This means that microorganisms containing target useful products of a certain quality or a culture solution containing useful products excreted into the culture solution can be continuously recovered, and industrial production becomes possible.

As described above, the present invention is a system for continuous large-scale cultivation of bacteria, yeast, and fungi, and employs a method in which a large amount of sterilized air is sprayed on a movable continuous foaming fixed bed together with the culture solution. The growth activity of microorganisms in the fixed bed is extremely high, and since the equipment is unitized, the production scale can be set arbitrarily. It is far superior to other mass production systems for specific useful metabolites, and has great value as a continuous useful product production system that has features such as being more compact and having a wide range of applications.

[Brief explanation of the drawing]

The figure is a schematic diagram of an embodiment of the microorganism continuous production apparatus according to the present invention. In the figure: 1... Upper reaction chamber 2... Spray nozzle 3... Air inlet 4... Microbial growth bed 5... Culture solution storage tank (lower reaction chamber) 6...・Power unit 7...Blower 8...Duct 9...Heat exchanger 10...Filter 11...Discharge duct 12...Culture solution storage tank 13...Exhaust Duct 14...Mist eliminator 15...Culture solution circulation pump 16...Communication pipe 17...Overflow pipe 18...Body external storage tank 19...Transfer pump 20...・・pH measuring device 21 ・・・chemical solution storage tank for pH adjustment 22 ・・・chemical solution injection pump 23 ・・culture solution discharge pump 24 ・・stock solution supply pump 25 ・・・high pressure spray water emission nozzle 26... High-pressure spraying liquid supply storage tank 27... High-pressure spray pump 28... Microbial liquid receiving tank 29... Microbial recovery tank 30... Recovery liquid transfer pump 31... Flowmeter patent applicant Sakai Engineering Co., Ltd. Agro Systems Co., Ltd.

Claims (1)

[Claims] 1. A reaction chamber for the growth of microorganisms and the production of useful products (biologically active substances such as enzymes and antibiotics), a growth bed for microorganisms (bacteria and fungi) that continuously passes through this reaction chamber and circulates endlessly, and Means for pumping and circulating a culture solution containing microorganisms for continuously supplying an atomized nutrient source to microorganisms inoculated on a bed in a reaction chamber, means for continuously supplying a large amount of air to the reaction chamber, and culturing. pH of liquid
It consists of auxiliary equipment such as a conditioning device and a device for collecting microorganisms in the growth bed. A continuous porous material is used as the growth bed for microorganisms, and this is supplemented with a large amount of air or oxygen as a nutrient source in the form of atomized water. A continuous microbial culture/separation device using a solid-state method, characterized in that the microorganisms are continuously supplied to the microorganisms, and only the amount of bacteria grown in an endless growth bed is recovered using high-pressure spray water.