KR102086617B1 - Method for the production of biomass of aerobic microorganisms - Google Patents

Abstract

The present invention relates to a method for producing biomass in aerobic microorganisms. More specifically, the present invention relates to a method for producing biomass in aerobic microorganisms, to be used for feed, food, and other purposes via biosynthesis methods of methane specifically different from natural gas in the field of microbiology. To this end, the method includes a process of culturing mixed microorganisms by continuously saturating gaseous hydrocarbons and initiators when removing accumulated biomass while supplying a nutrient medium under conditions of continuous chemostat biosynthesis.

Description

Method for the production of aerobic microorganisms {Method for the production of biomass of aerobic microorganisms}

The present invention relates to a method for producing biomass of aerobic microorganisms, and more particularly, to a method for producing biomass of aerobic microorganisms for feed, food and other purposes by the biosynthesis method of natural gas and other methane in the field of microbiology. have.

Currently, the production of methane nutrient proteins is reportedly produced by two companies, Calysta, California, USA and UniBio, Denmark. Except for these two, there is no active production of methane nutrient proteins worldwide, and only experimental techniques have been reported.

They were also used to recycle CH4 and O2 gas mixtures using a U-circuit in a pilot industrial machine capable of producing 10,000 tonnes of protein per year in Sbetly Yar, which operated from 1985 to 1994.

This technique is known as the know-how of the two companies, with the only difference being that they recycle the culture along the circuit and inject gas at the same time.

In other words, the recycling and mixing of the culture medium is carried out using a high speed mixer and pump, through which the culture medium is continuously circulated, and the microorganisms produced there are problems of stress, suppression and even deactivation. There was a problem of unintensive and unstable forms.

On the other hand, there are many prior art for the production of protein methane microorganisms, as follows.

1. WO2010069313A2 / 2010.06.24 Publication / U-shaped or nozzle U-loop fermenter and method of fermentation

2. EP0306466A2 / 1989.03.08 Publication / Methods and means for microbial production

3.DE2308087A1 / August 22, 1974 Publication / entry into force

4. EP0808910A2 / 1997.11.26 Publication / Apparatus and method for the production and use of iron ions produced by bacteria

5.EP0829534A2 / 1998.03.18 Public / gas-propelled fermentation system

6.US7579163 / 2011.03.02 Registration / Fermentation method

7.US20100035330 / 2010.02.11 Disclosure / Bioreactor

8. WO2003016460A1 / 2003.02.27 Disclosure / fermentation method

9. WO2014060778A1 / 2014.04.24 Publication / Fermentation apparatus and method of fermentation for protein production

Further known techniques are described in the book "Fermentation Devices for Microbiological Synthesis Processes (Author: A. Yu. Vinarov, L. S. Gordeev, A. A. Kukarenko, V. I. Panfilov.).

This technique pours a culture medium containing a culture of microorganisms capable of absorbing mineral salts and trace gaseous substrates (natural, liquefied gas) required for the essential activity of the microorganisms in a vessel that can operate at high pressure.

The circulation pump then provides a continuous circulation of the culture through a closed circuit, the sequence being circulated from the tank to the pump and then to a liquid jet ejector that sucks the mixed gas of oxygen and natural gas and finally to the next tank.

Here, the suction nozzle of the ejector is attached to the upper portion of the tank to recycle the mixture of natural gas and acid several times, and the clean quantity of natural gas and oxygen mixed in a certain ratio is also supplied to the inlet of the ejector.

At this time, the ratio of natural gas and oxygen should exclude the risk of explosion.

Thereafter, the culture medium is intensively and continuously circulated in the tank to discharge the medicine together with the culture solution from the liquid injector to continuously contact the pure oxygen and gas mixture.

However, such a device was installed in a biochemical plant in Nart-Kala city in 1972, but there was a problem in that it could not be actually operated due to the high risk of explosion and lack of technology.

On the other hand, a description of the method of growing microorganisms and the equipment for their execution is disclosed in Russian Patent No. 2021353, which is a water mineral nutrition that circulates between two zones for gas supply to microorganisms and consumption of introduced gases. Culturing the microorganisms in the medium.

At this time, the introduction of the feed gas is performed through the surface of the diffusion membrane while adjusting the rate of introduction of the gas in accordance with the growth rate of the microorganism by changing the partial pressure of each feed gas, the introduction of the gas is carried out individually through the surface of the plurality of membranes Can be.

The apparatus for carrying out this method comprises a culture unit and a gas delivery unit, the latter consisting of a group of hydrophobic membrane elements which form a cavity between their rows in communication with the gas source.

On the other hand, known methods of producing biomass include methods published by the Soviet Union for the study of protein material biosynthesis and the Institute of Microbiological Science and Technology (GDR), which produce a constant concentration of one of the elements of the nutrient medium during microbial cultivation. Nutrient medium containing a source of nitrogen, phosphorus, inorganic salts and methane as the carbon source, while culturing the methane oxidizing bacteria under non-sterile conditions, and maintaining the concentration of copper ions at 0.05 to 2 mg / l There is a technique.

Further, as a method for the production of aerobic microbial biomass, there is Russian Patent Publication No. 232488 (published on April 20, 2008).

The method includes culturing the microorganisms under conditions of continuously circulating the culture in a closed circuit and continuously separating and saturating with gaseous hydrocarbons and aeration agents when feeding the mineral nutrient medium and removing the accumulated biomass.

Here, the aeration agent is an aeration agent, and once removed from contact with the culture solution to maintain the saturation state.

In addition, saturation of gaseous hydrocarbons is carried out by repeated contact with the culture due to the recycling of gaseous hydrocarbons in the closed loop until complete dissolution.By producing biomass in this way, energy consumption for production is reduced and productivity is improved. You can.

This conventional microbial biomass production method will be described in more detail with reference to FIG. 1.

As shown in FIG. 1, first, a culture medium containing microorganisms capable of absorbing gaseous substrates and mineral salts and additives necessary for the biological activity of the microorganisms is placed in a container capable of operating at high pressure.

Thereafter, the circulation pump continuously circulates the culture solution from the tank to the pump and the liquid ejector in a closed loop to inhale a gas mixture of oxygen and natural gas and then continuously circulate into the tank.

At this time, the suction nozzle of the ejector is attached to the upper portion of the tank to recycle the mixture of natural gas and oxygen several times, and the clean quantity of natural gas and oxygen mixed in a specific ratio is also supplied to the inlet of the ejector.

At this time, the ratio of natural gas and oxygen should exclude the risk of explosion.

Thereafter, the culture medium in the tank is intensively and continuously circulated to discharge the medicament along with the culture medium from the liquid injector to continuously contact the pure oxygen and gas mixture, where the culture medium consumes oxygen and natural gas, and the carbon dioxide and biological Is made to release heat.

In addition, the content of oxygen and natural gas in the aeration agent is reduced to a certain value and the aeration agent is released from the tank when the device consumes up to 90-95% of the natural gas and oxygen.

This process is carried out continuously with constant supply of the culture to the tank and removal of the culture with the biomass of the accumulated microorganisms, with a constant discharge of the used aeration agent and heat.

However, the known disadvantages of this method are as follows.

1. The complexity of the design of the equipment and the many equipment components create operational reliability and risk of downtime.

2. The operation and maintenance cost of the equipment is high.

3. Continuous and continuous pumping of the culture is similar to the usual bubbling mixing method and causes stress on the microorganisms, thus reducing efficiency.

4. High energy costs.

5. Metabolites, including carbon dioxide, and unused air can be released into the atmosphere, causing environmental pollution.

Due to these problems, the need for a method of producing environmentally friendly biomass is being emphasized.

The present invention is to solve the above problems, an object of the present invention is to maximize the use of gas and by-products in the biomass biosynthesis process and biomass production method of aerobic microorganisms through eco-friendly fermenter through cooling system and air purification To provide.

Problems to be solved by the present invention are not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

Biomass production method of aerobic microorganism according to the present invention to solve the above problems, a biomass production method of aerobic microorganisms, gaseous hydrocarbon when supplying a nutrient medium under continuous chemostat biosynthesis conditions and remove the accumulated biomass And a method of culturing the mixed microorganisms by continuously saturating the initiator, and characterized in that mixing, aeration and saturation with gaseous hydrocarbons are performed by the mixing aeration head.

At this time, the mixing aeration head is characterized in that it has a structure for generating a vortex flow that rotates and rises around its own axis in the fermenter using the injection energy of the nutrient medium.

In addition, the nutrient medium mixed by continuously saturating the gaseous hydrocarbons and the initiator is characterized in that it is carried out in two stages (manufacturing and biosynthesis process step of the nutrient medium).

In this case, when the gaseous hydrocarbon and the initiator are saturated in the step of preparing the nutrient medium, the nutrient medium is cooled.

In addition, the nutrient medium saturated with gaseous hydrocarbons and the initiator is cooled as a coolant for controlling the temperature of the culture in the biosynthesis process step.

In addition, the mixed culture medium is characterized in that it further comprises a microbial strain using carbon dioxide as a carbon source, using hydrogen dissolved in the culture medium as an energy source.

At this time, when the carbon dioxide is excessive, the carbon dioxide is characterized in that it is used for photosynthesis of cyan spirulina or algae.

In addition, the initiator includes air, the gaseous hydrocarbon is characterized in that it comprises any methane and methane homologue.

Such a biomass production method of aerobic microorganism according to the present invention is energy-intensive in the biosynthesis process of protein compounds, there is an effect to provide an environmentally friendly method without stress on the microorganisms.

In addition, gas and biosynthetic by-products can be used to the maximum in the biosynthesis process, and in particular, in the early stage of fermentation, the nutrient medium is cooled before being fed to the fermentor, so that it is not only highly soluble but also can be used as a refrigerant of the cooling system. There is this.

In addition, the mixing aeration head using the injection energy of the nutrient medium produces an upward vortex of the culture solution, which has the effect of promoting efficient mixing and aeration of the culture solution.

Effects of the present invention are not limited to the above-mentioned effects, and other effects not mentioned will be clearly understood by those skilled in the art from the following description.

1 is a view showing the prior art of the biomass production method,
2 is a view schematically showing a fermentor apparatus according to the biomass production method of aerobic microorganisms of the present invention,
Figure 3 schematically shows a multi-nozzle injector according to the method for producing biomass of aerobic microorganisms of the present invention.

As the present invention allows for various changes and numerous embodiments, particular embodiments will be illustrated in the drawings and described in detail in the written description.

However, this is not intended to limit the present invention to specific embodiments, it should be understood to include all modifications, equivalents, and substitutes included in the spirit and scope of the present invention. In describing the drawings, similar reference numerals are used for similar elements.

When a component is said to be "connected" or "connected" to another component, it may be directly connected to or connected to that other component, but it may be understood that another component may be present in the middle. Should be. On the other hand, when a component is said to be "directly connected" or "directly connected" to another component, it should be understood that there is no other component in between.

The terminology used herein is for the purpose of describing particular example embodiments only and is not intended to be limiting of the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In this application, the terms "comprise" or "have" are intended to indicate that there is a feature, number, step, operation, component, part, or combination thereof described in the specification, and one or more other features. It is to be understood that the present invention does not exclude the possibility of the presence or the addition of numbers, steps, operations, components, components, or a combination thereof.

Hereinafter, with reference to the accompanying drawings, it will be described in detail a preferred embodiment of the present invention. Hereinafter, the same reference numerals are used for the same components in the drawings, and duplicate descriptions of the same components are omitted.

Hereinafter, with reference to the accompanying drawings, it will be described embodiments of the present invention. Like reference numerals in the drawings denote like elements. In describing the present invention, detailed descriptions of related well-known functions or configurations are omitted in order not to obscure the subject matter of the present invention.

2 is a view schematically showing a fermentor apparatus according to the biomass production method of aerobic microorganisms of the present invention, Figure 3 is a view schematically showing a multi-nozzle injector according to the biomass production method of aerobic microorganisms of the present invention.

2 to 3, the biomass production method of aerobic microorganism according to the present invention is a biomass production method of aerobic microorganisms, supplying a nutrient medium under continuous chemostat biosynthesis conditions and accumulates the biomass accumulated The method includes culturing the mixed microorganisms by continuously saturating the gaseous hydrocarbons and the initiator upon removal, and characterized in that mixing, aeration and saturation are performed with the gaseous hydrocarbons by the mixing aeration head.

At this time, the nutrient medium is a nutrient for microorganisms, and may include elements necessary for growth, trace elements, vitamins and growth promoters.

At this time, the rate of supplying the nutrient medium corresponds to the selection rate of the culture solution from the fermenter, the optimal number of generation of microorganisms introduced into the mixed production culture consisting of bacteria using carbon dioxide as a carbon source, as well as bacteria of methane and methane homologs This can be determined by considering.

In addition, the fermenter can be enriched with the culture liquid by venting methane and air using the multi-nozzle injector 14.

Here, the mixing aeration head may serve as a pipe pump circuit for the recirculation of the culture as well as the role of a high speed stirrer for aeration and mixing.

At this time, the mixing aeration head is characterized in that it has a structure for generating a vortex flow that rotates and rises around its own axis in the fermenter using the injection energy of the nutrient medium.

In addition, the nutrient medium mixed by continuously saturating the gaseous hydrocarbons and the initiator is characterized in that it is carried out in two stages (manufacturing and biosynthesis process step of the nutrient medium).

In this case, when the gaseous hydrocarbon and the initiator are saturated in the step of preparing the nutrient medium, the nutrient medium is cooled.

In addition, the nutrient medium saturated with gaseous hydrocarbons and the initiator is cooled as a coolant for controlling the temperature of the culture in the biosynthesis process step.

In addition, the mixed culture medium is characterized in that it further comprises a microbial strain using carbon dioxide as a carbon source, using hydrogen dissolved in the culture medium as an energy source.

At this time, when the carbon dioxide is excessive, the carbon dioxide is characterized in that it is used for photosynthesis of cyan spirulina or algae.

In addition, the initiator includes air, the gaseous hydrocarbon is characterized in that it comprises any methane and methane homologue.

When explaining the process of the biomass production method of aerobic microorganism according to the present invention.

First, a nutrient medium made up of salts of trace elements supplied from the dispenser 2 is supplied to the mixer 4 and sent to a supply tank 5 which is mixed with water supplied from the sterilizer.

Thereafter, the nutrient medium is sent to the nutrient medium sterilization device 6, and sent to the final forming device 8 for final forming the nutrient medium, in which the nutrient medium is generated in the sterilizing device 7. It is mixed with the printing solution. Here, an aqueous solution based on sterilized water, salts and phosphorus, nitrogen, potassium and magnesium mixed solutions may be prepared.

The obtained nutrient medium is then fed to a cooling system to predissolve the methane and air filtered through the gas filter 12 in the nutrient medium.

Thereafter, the filtered methane and air are supplied to the nutrient medium, and the methane obtained by separating the gas of carbon dioxide and the methane not used in the fermentor with the gas separation filter 13 is also supplied.

In addition, the pre-dissolving device 10 may serve as a cooler, so that the dissolution of the filtered methane and air can be easily performed.

Thereafter, the methane and the air are saturated with the pressurized pump 11, and the cooled nutrient medium is supplied to the lower portion of the fermentor 1. At this time, the methane and air filtered through the gas filter 12 is supplied to the fermentor.

At this time, the pressure pump is a gas outlet is formed in the distal end portion, is formed in a flat shape, similar to the cross section of the plane as a whole serves to maintain the flow of the culture solution from the mixing aeration head.

In addition, the water injected in the mixed culture form is to be supplied once to the fermentor after the partial feeding of the nutrient medium.

Thereafter, the excessively generated carbon dioxide is separated from the filter filter 13 and then supplied to the photosynthesis reactor 15 for culturing cyan spirulina or algae, and the biomass separated from the separator 16 is dried in the dryer 17. Then, the tableting device 18 to be purified.

Thereafter, the biomass of the aerobic microorganisms grown is separated from the filtrate in the centrifuge (19) and sent to the biomass dryer (21), and the biomass separated from the filtrate filter (20) is also transferred to the biomass dryer (21). Is sent.

The completely dried biomass is then sent to granulator 22 for packaging.

Finally, the component analyzer 9 analyzes the chemical composition for reuse in the circulating water supply system and is sent to the water treatment apparatus 3 for purification.

Meanwhile, as shown in FIG. 3, the multi-nozzle injector 14 according to the present invention will be described as follows.

The multi-nozzle injector is formed on the wall of the fermentor, and the nozzles oriented along the streamline not only supply abundant gas to the culture, but also support vortex movement of the culture.

Here, the vortex movement of the culture is produced by the mixing aeration head formed at the bottom of the multi-nozzle injector, which allows 20% of the injected nutrient medium to be continuously mixed with 80% of the culture in the fermentor. In addition, the culture medium along the inner wall of the fermenter to create a vortex flow that rotates and rises about its own axis.

Such a biomass production method, fermenter and multi-nozzle injector of the aerobic microorganism according to the present invention has the following effects.

The nutrient medium, which is continuously saturated with methane and air and cooled at two stages before the fermentation and during the biosynthesis process, can provide a nearly complete source of carbon and energy to the microorganisms through the culture.

In addition, the heterotrophic bacteria strains are used to utilize the metabolites accumulated in the biosynthesis process, and the mixed culture medium thus configured can solve many problems related to foaming, high concentration, and excessive oxidation, and does not stress microorganisms. Of course, less waste can provide an environmentally friendly and stable way.

In addition, excess carbon dioxide can be separated and used for photosynthesis of cyan spirulina or algae.

The cooled nutrient medium can also be used as a refrigerant in the system for monitoring and controlling the temperature of the culture.

In addition, the protein concentrates obtained through the process passed the standard test for animal and poultry feeds of various types and ages, and the average of 1 ton of protein per 20 tons of protein, vitamins, essential amino acids and trace elements among the feed ingredients. Can be balanced by proportions.

In addition, after purifying bacterial biomass from RNA components, proteins suitable for use in foods can be obtained.

In addition, the RNA component isolated during the purification of biomass accounts for 15% of the biomass, and may be suitable as a raw material and a production ingredient required in the pharmaceutical and perfume, medical and food industries.

The best embodiments have been disclosed in the drawings and the specification. Herein, specific terms have been used, but they are used only for the purpose of illustrating the present invention and are not intended to limit the scope of the present invention as defined in the meaning or claims. Therefore, those skilled in the art will understand that various modifications and equivalent other embodiments are possible from this. Therefore, the true technical protection scope of the present invention will be defined by the technical spirit of the appended claims.

1 ... fermenter 2 ... dispenser
3 ... water treatment unit 4 ... mixer
5 ... supply tank 6 ... nutrient medium sterilizer
7 ... Sterilizer 8 ... Final Forming Apparatus
9 ... component analyzer 10 ... predissolution apparatus
11 ... pressurized pump 12 ... gas filter
13 ... filtration filter 14 ... multi-nozzle injector
15 ... photosynthesis reactor 16 ... separator
17 ... dryer 18 ... tableting device
19 ... centrifuge 20 ... filtrate filter
21 ... Biomass Dryer 22 ... Granulator

Claims (8)

A method for producing biomass of aerobic microorganisms, the method comprising culturing mixed microorganisms by continuously saturating gaseous hydrocarbons and initiators when supplying a nutrient medium under continuous chemostat biosynthesis conditions and removing accumulated biomass, and mixed aeration In the biomass production method of aerobic microorganisms characterized in that mixing, aeration and saturation into gaseous hydrocarbons by the head,
The saturation of the gaseous hydrocarbons and initiators is successively saturated so that the mixed nutrient medium is carried out during the production and biosynthesis process steps of the nutrient medium,
In the manufacturing step of the nutrient medium to pre-dissolve the gaseous hydrocarbons and initiators filtered by the gas filter 12 through the pre-dissolution apparatus 10 in the nutrient medium, when the gaseous hydrocarbons and initiators are saturated, the nutrient medium It is characterized in that to be supplied to the lower portion of the fermenter (1) in the first cooled state, characterized in that used in the biosynthetic process step as a coolant for controlling the temperature of the culture,
The mixed culture medium is characterized in that it further comprises a microbial strain using carbon dioxide as a carbon source, using hydrogen dissolved in the culture medium as an energy source,
When the carbon dioxide is excessive, the carbon dioxide is a biomass production method of aerobic microorganisms, characterized in that it is used for photosynthesis of cyan spirulina or algae. The method of claim 1,
The mixing aeration head,
The biomass production method of aerobic microorganisms having a structure for generating a vortex flow that rotates and rises about its own axis in the fermenter using the energy of the nutrient medium.
delete delete delete delete delete The method of claim 1,
Wherein said initiator comprises air, and said gaseous hydrocarbons comprise any methane and a methane homologue.

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