WO2021033950A1

WO2021033950A1 - Method for producing biomass of aerobic microorganism

Info

Publication number: WO2021033950A1
Application number: PCT/KR2020/010107
Authority: WO
Inventors: 조윤경; 알렉산드로비치 글루히흐세르게이; 페트로비치 템류코브안드레이; 세르게예비치 글루히흐아르카디이; 안드레예브나 템류코바베로니카
Original assignee: 주식회사 바이오프린
Priority date: 2019-08-16
Filing date: 2020-07-31
Publication date: 2021-02-25
Also published as: KR102086617B1

Abstract

The present invention relates to a method for producing biomass of aerobic microorganisms and, more specifically, is concerned with a method for producing biomass of aerobic microorganisms for feeds, foods, and other purposes through biosynthesis of methane different from natural gas in the microbiology field.

Description

Biomass production method 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 biosynthetic methods of natural gas and other methane in the field of microbiology. have.

Currently, production of methanol protein is reported to be produced only by two companies, Calysta of California and UniBio of Denmark. Excluding these two locations, there is no active production of methanol proteins worldwide, and only experimental techniques have been reported.

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

This technology is known for the know-how of the two companies, the only difference is that they recycle the culture broth along this circuit and simultaneously inject gas.

In other words, recycling and mixing of the pill solution are performed using a high-speed mixer and pump, through which the culture solution is continuously circulated, and the microorganisms produced here have problems of being stressed, inhibited and even inactivated. Was not intensive and had an unstable form.

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

1. WO2010069313A2 / 2010.06.24 Publication / U-shape or nozzle U-loop fermentation tank and method of fermentation

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

3. DE2308087A1 / 1974.08.22 Disclosure / Entry into force procedure

4. EP0808910A2 / Publication on November 26, 1997 / 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 / Entry into force method

7. US20100035330 / 2010.02.11 Public / Bioreactor

8. WO2003016460A1 / published on February 27, 2003 / fermentation method

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

In addition, other known techniques are described in the book "Fermentation apparatus for microbiological synthesis (authors: A.Yu. Vinarov, L.S. Gordeev, A.A. Kukarenko, V.I. Panfilov.)", and are as follows.

This technique pours a culture solution containing a culture of microorganisms capable of absorbing traces of gaseous substrates (natural, liquefied gases) and mineral salts necessary for the essential activity of microorganisms in a vessel capable of operating at high pressure.

Subsequently, the circulation pump provides continuous circulation of the culture medium through a closed circuit, and the sequence is a liquid injection discharger that sucks a mixed gas of oxygen and natural gas from the tank to the pump, and finally circulates to the next tank.

Here, the suction nozzle of the ejector is attached to the top of the tank so that the mixture of natural gas and acid can be recycled several times, and the clean quantity of natural gas and oxygen mixed in a certain ratio are also supplied to the inlet of the ejector.

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

Thereafter, the QOL is intensively and continuously circulated inside the tank to discharge the drug together with the culture solution from the liquid spraying device 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 is a problem that it is not actually operated because of high risk of explosion and lack of technology.

On the other hand, a technique for cultivating microorganisms and equipment for their implementation is published in Russian Patent No. 2021353, which is a method for supplying gas to microorganisms and circulating water mineral nutrients between the two zones for consumption of the introduced gas. It includes culturing the microorganism in a medium.

At this time, the introduction of the supply gas is performed through the surface of the diffusion film while controlling the introduction rate of the gas according to the growth rate of the microorganism by changing the partial pressure of each supply gas, and the introduction of the gas is performed individually through the surfaces of the multiple membranes. Can be.

An 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 forming a cavity between their rows in communication with a gas source.

Meanwhile, a known method for producing biomass is a method published by the USSR (Protein Biosynthesis Research Institute) and Microbial Science and Technology Research Institute (GDR) of the former Soviet Union, and this technology is one of the elements of the nutrient medium in the microbial culture process. A nutrient medium containing a source of nitrogen, phosphorus, inorganic salts and methane as a carbon source while maintaining a constant concentration includes cultivation of methane oxidizing bacteria under non-sterile conditions, and the concentration of copper ions in the culture process is 0.05 to 2 mg/1 There is a skill to keep it as.

In addition, as a method for the production of aerobic microbial biomass, there is Russian Patent Publication No.2322488 (published on April 20, 2008).

This method includes culturing the microorganisms under conditions of continuously circulating the culture medium in a closed circuit when supplying a mineral nutrient medium and removing accumulated biomass, and continuously separating and saturating with gaseous hydrocarbons and an aerosol.

Here, the aeration agent is an aeration agent, and it is removed after contacting the culture solution once to maintain saturation.

In addition, the saturation of gaseous hydrocarbons is carried out by repeatedly contacting the culture medium due to the recycling of gaseous hydrocarbons in a closed loop until completely dissolved.If biomass is produced in this way, energy consumption for production is reduced and productivity is improved. I can make it.

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

As shown in FIG. 1, first, a culture solution of a microorganism capable of absorbing a gaseous substrate and a culture solution containing mineral salts and additives necessary for the life activity of the microorganism are 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 suck a gas mixture of oxygen and natural gas, and then continuously circulate it to the tank.

At this time, the suction nozzle of the ejector is attached to the top of the tank to recirculate the mixture of natural gas and oxygen 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.

Thereafter, the culture solution inside the tank is intensively and continuously circulated to discharge the drug along with the culture solution in the liquid injection device to make the pure oxygen and gas mixture in continuous contact, where the culture solution consumes oxygen and natural gas and consumes carbon dioxide and biological It is made to release heat.

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

This process is carried out continuously while constantly supplying the culture solution to the tank, removing the culture solution with the accumulated biomass of microorganisms, and constantly discharging the used aeration agent and heat.

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

1. Due to the complexity of the design of the equipment and the large number of equipment parts, there is a risk of operational reliability and disruption.

2. Equipment operation and maintenance costs are high.

3. Continuous and continuous pumping of the culture medium is the same as the general bubbling mixing method and causes stress on microorganisms, resulting in reduced efficiency.

4. High energy cost.

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

Due to these problems, the necessity of a method for producing eco-friendly biomass is being emphasized.

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

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

In order to solve the above-described problems, the method for producing biomass of aerobic microorganisms according to the present invention is a method for producing biomass of aerobic microorganisms, and when supplying a nutrient medium under continuous chemostat biosynthesis conditions and removing accumulated biomass, gaseous hydrocarbons And a method of culturing the mixed microorganisms by continuously saturating the initiator, and mixing, aeration, and saturation are performed with gaseous hydrocarbons by a mixing aeration head.

At this time, the mixed aeration head is characterized in that it has a structure to generate a vortex flow that rotates and rises around its own axis in the fermentor by using the injected energy of the nutrient medium.

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

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

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

In addition, the mixed culture solution is characterized in that it further includes a microorganism strain using carbon dioxide as a carbon source and hydrogen dissolved in the culture solution as an energy source.

In this case, when carbon dioxide is excessive, the carbon dioxide is used for photosynthesis of blue-green spirulina algae.

In addition, the initiator contains air, and the gaseous hydrocarbon is characterized in that it contains any methane and methane homologues.

The biomass production method of aerobic microorganisms according to the present invention is energy-intensive in the process of biosynthesis of protein compounds, and has an effect of providing an eco-friendly method without stress on microorganisms.

In addition, it allows the maximum use of gases and biosynthetic by-products during the biosynthesis process, and in particular, since the nutrient medium is cooled before being supplied to the fermenter in the initial stage of fermentation, it has high solubility and can be used as a refrigerant for the cooling system. There is this.

In addition, there is an effect of generating a rising vortex of the culture solution by the mixing aeration head using the injection energy of the nutrient medium, thereby promoting efficient mixing and ventilation of the culture solution.

The effects of the present invention are not limited to the effects mentioned above, 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 method for producing biomass of aerobic microorganisms of the present invention;

3 is a schematic view showing a multi-nozzle injector according to the method for producing biomass of aerobic microorganisms of the present invention.

In the present invention, various modifications may be made and various embodiments may be provided, and specific embodiments will be illustrated in the drawings and described in detail.

However, this is not intended to limit the present invention to a specific embodiment, it is to be understood to include all changes, equivalents, and substitutes included in the spirit and scope of the present invention. In describing each drawing, similar reference numerals have been used for similar elements.

When a component is referred to as being "connected" or "connected" to another component, it is understood that it may be directly connected or connected to the other component, but other components may exist in the middle. Should be. On the other hand, when a component is referred to as being "directly connected" or "directly connected" to another component, it should be understood that there is no other component in the middle.

The terms used in the present application are only used to describe specific embodiments, and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In the present application, terms such as "comprise" or "have" are intended to designate the presence of features, numbers, steps, actions, components, parts, or combinations thereof described in the specification, but one or more other features. It is to be understood that the presence or addition of elements or numbers, steps, actions, components, parts, or combinations thereof, does not preclude in advance.

Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to the accompanying drawings. Hereinafter, the same reference numerals are used for the same elements in the drawings, and duplicate descriptions for the same elements are omitted.

Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. The same reference numerals in each drawing indicate the same members. In describing the present invention, detailed descriptions of related 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 method for producing biomass of aerobic microorganisms of the present invention, and FIG. 3 is a view schematically showing a multi-nozzle injector according to the method for producing biomass of aerobic microorganisms of the present invention.

As shown in Figs. 2 to 3, the method for producing biomass of aerobic microorganisms according to the present invention is a method for producing biomass of aerobic microorganisms, in which a nutrient medium is supplied under continuous chemostat biosynthesis conditions, and the accumulated biomass is It includes a method of culturing mixed microorganisms by continuously saturating gaseous hydrocarbons and an initiator when removed, characterized in that mixing, aeration and saturation are performed with gaseous hydrocarbons by a mixing aeration head.

At this time, the nutrient medium is a nutrient for microorganisms, and may contain 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 broth from the fermentor, and the optimal number of generations of microorganisms entering the mixed production culture composed of methane and methane homologs as well as bacteria using carbon dioxide as a carbon source Can be determined by taking into account.

In addition, the fermenter may be enriched with methane and air by using a multi-nozzle injector 14.

Here, the mixing aeration head may serve not only as a high-speed stirrer for ventilation and mixing, but also as a pipe pump circuit for recirculation of the culture solution.

Accordingly, the process of the method for producing biomass of aerobic microorganisms according to the present invention will be described as follows.

First, the nutrient medium consisting of salts of trace elements supplied from the dispenser 2 is supplied to the mixer 4 and sent to the supply tank 5 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 formation of the nutrient medium. In the final forming device, the nutrient medium is produced in the sterilizing device 7 It is mixed with the printing solution. Here, an aqueous solution based on a mixed solution of sterilized water, salt and phosphorus, nitrogen, potassium and magnesium may be prepared.

Thereafter, the obtained nutrient medium is supplied to a cooling system, and methane and air filtered through the gas filter 12 are pre-dissolved in the nutrient medium.

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

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

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

At this time, the pressure pump has a gas outlet formed at the end thereof, and is formed in a flat shape similar to the cross-section of an airplane wing as a whole to maintain the flow of the culture solution from the mixing aeration head.

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

Thereafter, the excess carbon dioxide is separated by the filter filter 13 and then supplied to the photosynthetic reactor 15 for culturing the cyan spirulina algae, and the biomass separated by the separator 16 is dried in the dryer 17. , To be purified in a tableting device (18).

Thereafter, the biomass of the proliferated aerobic microorganisms is separated from the filtrate in the centrifuge 19 and sent to the biomass dryer 21, and the biomass separated by the filtrate filter 20 is also transferred to the biomass dryer 21. Sent.

Thereafter, the completely dried biomass is sent to the granulator 22 to be packaged.

Finally, the component analyzer 9 analyzes the chemical composition for reuse in the circulation water supply system, and is sent to the water treatment device 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 stream line not only supply abundant gas to the culture medium, but also support the vortex motion of the culture medium.

Here, the vortex motion of the culture medium is generated by the mixing aeration head formed under the multi-nozzle injector, and the mixing aeration head allows 20% of the injected nutrient medium to be continuously mixed with 80% of the culture medium in the fermentor. In this case, the culture broth rotates and rises around its own axis along the inner wall of the fermentor to create a vortex flow.

The biomass production method of aerobic microorganisms, fermentor and multi-nozzle injector according to the present invention have the following effects.

The nutrient medium, which is continuously saturated with methane and air and cooled in the two stages of the pre-fermentation stage and the biosynthetic process stage, can almost completely provide carbon and energy sources to microorganisms through the culture medium.

In addition, it includes strains of heterotrophic bacteria to use metabolites accumulated in the biosynthesis process, and can solve many problems related to foaming, high concentration, and excessive oxidation by such a mixed culture solution, and does not put stress on microorganisms. Of course, it can provide an eco-friendly and stable method with less waste.

In addition, it can be used for photosynthesis of blue-green spirulina algae by separating excess carbon dioxide.

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

In addition, the protein concentrate obtained through the process passed the standard test for animal and poultry feeds of various types and ages, and among the ingredients of the feed, protein, vitamins, essential amino acids and trace elements, etc. You can balance it with proportions.

Further, after purification of the bacterial biomass from the RNA component, a protein suitable for use in food can be obtained.

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

Optimal embodiments have been disclosed in the drawings and specification. Here, specific terms have been used, but these are only used for the purpose of describing the present invention, and are not used to limit the meaning or the scope of the present invention described in the claims. Therefore, those of ordinary skill in the art will understand that various modifications and equivalent other embodiments are possible therefrom. Accordingly, the true technical scope of the present invention should be determined by the technical spirit of the appended claims.

Claims

As a method of producing biomass of aerobic microorganisms, it includes a method of culturing mixed microorganisms by continuously saturating gaseous hydrocarbons and an initiator when supplying a nutrient medium under continuous chemostat biosynthesis conditions and removing accumulated biomass, and mixing aeration A method for producing biomass of aerobic microorganisms, characterized in that mixing, aeration and saturation are performed with gaseous hydrocarbons by a head.
The method of claim 1,

The mixed aeration head,

A method for producing biomass of aerobic microorganisms, characterized in that it has a structure that generates a vortex flow that rotates and rises around its own axis in a fermentor by using the injection energy of the nutrient medium.
The method of claim 1,

A method for producing biomass of aerobic microorganisms, characterized in that the nutrient medium mixed by continuously saturating gaseous hydrocarbons and an initiator is carried out in two steps (a manufacturing step of a nutrient medium and a biosynthetic process step).
The method of claim 3,

When the gaseous hydrocarbons and the initiator are saturated in the step of preparing the nutrient medium, the nutrient medium is cooled.
The method of claim 4,

A method for producing biomass of aerobic microorganisms, characterized in that the nutrient medium saturated and cooled with gaseous hydrocarbons and an initiator is used as a coolant for controlling the temperature of the culture medium in the biosynthetic process step.
The method of claim 1,

The mixed culture medium uses carbon dioxide as a carbon source, and a microbial strain that uses hydrogen dissolved in the culture medium as an energy source further comprises a biomass production method of aerobic microorganisms.
The method of claim 6,

When carbon dioxide is excessive, the carbon dioxide is used for photosynthesis of blue-green spirulina algae.
The method of claim 1,

The initiator comprises air, and the gaseous hydrocarbon is a method for producing biomass of aerobic microorganisms, characterized in that it contains any methane and methane analogs.