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CN115927766B - A culture condition control method and system for improving the optical attenuation performance of microorganisms - Google Patents

A culture condition control method and system for improving the optical attenuation performance of microorganisms Download PDF

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CN115927766B
CN115927766B CN202310152604.7A CN202310152604A CN115927766B CN 115927766 B CN115927766 B CN 115927766B CN 202310152604 A CN202310152604 A CN 202310152604A CN 115927766 B CN115927766 B CN 115927766B
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microorganism
dipole
particle size
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CN115927766A (en
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胡以华
王新宇
杨星
顾有林
石亮
王磊
陈曦
梁振宇
陈友龙
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National University of Defense Technology
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Abstract

本发明给出了一种提升微生物光学衰减性能的培养条件控制方法和系统,属于微生物材料技术领域,该方法包括:获取影响微生物光学衰减性能的目标波段;确定微生物在目标波段内的颗粒粒径优化目标、培养基液固比、培养环境条件、无机盐添加量和培养基上孢子接种量;获取微生物在目标波段内的脂质含量和蛋白质含量,以确定培养基氮源和碳源的种类和含量以及微生物孢子干燥方法;按照颗粒粒径优化目标、培养基液固比、培养环境条件、无机盐添加量、培养基上孢子接种量、培养基氮源和碳源的种类和含量以及微生物孢子干燥方法,培养所述微生物。本发明在保证微生物孢子产量和代谢物产量的前提下,提升了微生物光学衰减性能,能够满足宽波段光学衰减需求。

The invention provides a culture condition control method and system for improving the optical attenuation performance of microorganisms, which belongs to the technical field of microbial materials. The method includes: obtaining a target waveband that affects the optical attenuation performance of microorganisms; determining the particle size of microorganisms in the target waveband Optimize the target, medium liquid-solid ratio, culture environment conditions, inorganic salt addition amount and spore inoculation amount on the medium; obtain the lipid content and protein content of microorganisms in the target band to determine the type of nitrogen source and carbon source of the medium and content and microbial spore drying method; according to the particle size optimization target, medium liquid-solid ratio, culture environment conditions, inorganic salt addition amount, spore inoculum amount on the medium, the type and content of nitrogen source and carbon source in the medium, and microbial The spore drying method, culturing the microorganisms. Under the premise of ensuring the yield of microbial spores and metabolites, the present invention improves the optical attenuation performance of microorganisms, and can meet the requirement of wide-band optical attenuation.

Description

Culture condition control method and system for improving optical attenuation performance of microorganisms
Technical Field
The invention belongs to the technical field of microbial materials, and particularly relates to a culture condition control method and system for improving optical attenuation performance of microorganisms.
Background
In recent years, the search for a high-efficiency extinction material with excellent optical attenuation performance has become a research hotspot for students at home and abroad. The manual preparation of microbial materials as a new direction of research and development of extinction materials has been demonstrated to have good optical attenuation properties over all-optical bands. Compared with the inorganic materials used by the traditional extinction materials, one of the advantages of artificially preparing the microbial materials as the extinction materials is that the microbial materials have various forms, the various forms provide favorable conditions for realizing broadband optical attenuation, and the microbial materials have rich components and high optical absorption capacity. However, the existing microorganism culture method cannot realize targeted control on the optical attenuation performance of microorganisms, and the prepared optical attenuation performance of microorganisms cannot meet the requirement of wide-band optical attenuation.
Disclosure of Invention
The invention aims at providing a culture condition control method for improving the optical attenuation performance of microorganisms, which improves the optical attenuation performance of microorganisms on the premise of ensuring the yield of spores of the microorganisms and can meet the broadband optical attenuation requirement.
The second object of the present invention is to provide a culture condition control system for improving the optical attenuation performance of microorganisms.
In order to achieve one of the above purposes, the present invention is implemented by the following technical scheme:
a culture condition control method for improving optical attenuation performance of microorganisms, the culture condition control method comprising:
s1, acquiring a target wave band affecting the optical attenuation performance of microorganisms;
s2, determining a particle size optimization target of the microorganism in the target wave band, a liquid-solid ratio of a culture medium, culture environment conditions, an inorganic salt addition amount and a spore inoculation amount on the culture medium;
s3, obtaining the lipid content and the protein content of the microorganism in the target wave band so as to determine the types and the contents of a nitrogen source and a carbon source of the culture medium and a microorganism spore drying method;
and S4, optimizing targets, a culture medium liquid-solid ratio, culture environment conditions, inorganic salt addition amount, spore inoculation amount on a culture medium, types and contents of a nitrogen source and a carbon source of the culture medium and a microbial spore drying method according to the particle sizes of the particles, and culturing the microorganisms.
Further, the specific implementation process of the step S2 includes:
s21, acquiring microorganisms with different particle sizes;
s22, obtaining extinction sections corresponding to microorganisms with different particle sizes in the target wave band;
s23, selecting the particle size corresponding to the maximum extinction section from extinction sections corresponding to microorganisms with different particle sizes in the target wave band as a particle size optimization target of the microorganisms in the target wave band;
s24, judging the transformation trend between the optical attenuation of the microorganism and the particle size of the particles in the target wave band according to the extinction cross sections corresponding to the microorganisms with different particle sizes in the target wave band so as to determine the liquid-solid ratio of the culture medium, the culture environment condition, the addition amount of inorganic salt and the spore inoculation amount on the culture medium of the microorganisms;
the transformation trend is: the microbial optical attenuation performance increases as the particle size of the microbial particles increases; alternatively, the microbial optical attenuation performance decreases as the microbial particle size increases.
Further, in the step S22, the specific process of obtaining the extinction cross section corresponding to the microorganism with different particle sizes in the target band includes:
step S221, respectively measuring the reflection spectrums of the microorganisms with different particle sizes in the target wave band to determine the reflection phase shift of the microorganisms with different particle sizes in the target wave band;
step S222, calculating complex refractive indexes corresponding to the microorganisms with different particle sizes in the target wave band according to the reflection spectrums and the corresponding reflection phase shifts of the microorganisms with different particle sizes in the target wave band;
step S223, according to the complex refractive index, calculating the dipole numbers corresponding to the microorganisms with different particle sizes in the target wave band and the dipole moment of each dipole;
and step 224, calculating extinction sections corresponding to microorganisms with different particle sizes in the target wave band according to the dipole number and the dipole moment of each dipole.
Further, in the step S223, the dipole moment of each dipole is:
wherein,,P i is the firstiDipole moments of the individual dipoles;α i is the firstiThe polarizability of the individual dipoles;is the firstiIncident field electric field strength of the individual dipoles; />To divide byiThe sum of the scattered field intensities generated by the remaining dipoles outside the individual dipoles;i=1,2,…,MMis the number of dipoles.
Further, in the step S224, the extinction cross section of the microorganism is:
wherein,,Cr j is of particle diameter ofr j Is a microbial extinction cross section;is the firstiIncident field electric field strength of individual dipoles +.>Complex conjugate of (a); lambda is the wavelength of the incident electromagnetic wave;j=1,2,…,NNis the particle size number of the particles.
Further, the specific implementation process of the step S3 includes:
s31, setting constraint conditions of the content of each component in the microorganism;
the components comprise proteins, lipids, nucleic acids and other microelements;
step S32, obtaining the average absorbance of each component on the target wave band so as to determine the average absorbance function of the microorganism on the target wave band;
s33, constructing a composition optimization model according to constraint conditions of the content of each composition and the average absorbance function so as to determine the lipid content and the protein content of the microorganism on the target wave band;
step S34, determining the types and the respective corresponding contents of nitrogen sources and carbon sources of a culture medium according to the lipid content of the microorganism on the target wave band;
and step S35, determining a microbial spore drying method according to the protein content of the microorganism on the target wave band.
In order to achieve the second purpose, the invention adopts the following technical scheme:
a culture condition control system for enhancing optical attenuation performance of microorganisms, the culture condition control system comprising:
the first acquisition module is used for acquiring a target wave band affecting the optical attenuation performance of the microorganism;
the determining module is used for determining a particle size optimization target of the microorganism in the target wave band, a liquid-solid ratio of a culture medium, culture environment conditions, an inorganic salt addition amount and a spore inoculation amount on the culture medium;
a second acquisition module for acquiring lipid content and protein content of the microorganism in the target wave band so as to determine the type and content of a nitrogen source and a carbon source of the culture medium and a microorganism spore drying method;
and the culture module is used for optimizing targets, a culture medium liquid-solid ratio, culture environment conditions, inorganic salt addition amount, spore inoculation amount on the culture medium, types and contents of a nitrogen source and a carbon source of the culture medium and a microbial spore drying method according to the particle size of the particles to culture the microorganisms.
Further, the determining module includes:
the first acquisition submodule is used for acquiring microorganisms with different particle sizes;
the second acquisition submodule is used for acquiring extinction sections corresponding to microorganisms with different particle sizes in the target wave band;
selecting a submodule, namely selecting the particle size corresponding to the maximum extinction cross section from extinction cross sections corresponding to microorganisms with different particle sizes in the target wave band as a particle size optimization target of the microorganisms in the target wave band;
the judging submodule is used for judging the transformation trend between the optical attenuation of the microorganism in the target wave band and the particle size of the particles according to the extinction cross sections corresponding to the microorganisms with different particle sizes in the target wave band so as to determine the liquid-solid ratio of the culture medium of the microorganisms, the culture environment condition, the addition amount of inorganic salt and the spore inoculation amount on the culture medium;
the transformation trend is: the microbial optical attenuation performance increases as the particle size of the microbial particles increases; alternatively, the microbial optical attenuation performance decreases as the microbial particle size increases.
Further, the second obtaining submodule includes:
the measuring submodule is used for respectively measuring the reflection spectrums of the microorganisms with different particle sizes in the target wave band so as to determine the reflection phase shift of the microorganisms with different particle sizes in the target wave band;
the first calculating submodule is used for calculating complex refractive indexes corresponding to the microorganisms with different particle sizes in the target wave band according to the reflection spectrums and the corresponding reflection phase shifts of the microorganisms with different particle sizes in the target wave band;
the second calculation submodule is used for calculating the dipole number and the dipole moment of each dipole corresponding to the microorganisms with different particle sizes in the target wave band according to the complex refractive index;
and the third calculation sub-module is used for calculating extinction sections corresponding to microorganisms with different particle sizes in the target wave band according to the dipole number and the dipole moment of each dipole.
Further, the second obtaining module includes:
the setting submodule is used for setting constraint conditions of the content of each component in the microorganism;
the components comprise proteins, lipids, nucleic acids and other microelements;
a third obtaining submodule, configured to obtain average absorbance of each component on the target band, so as to determine an average absorbance function of the microorganism on the target band;
the constructing submodule is used for constructing a composition optimizing model according to the constraint conditions of the content of each composition component and the average absorbance function so as to determine the lipid content and the protein content of the microorganism on the target wave band;
a first determining submodule, configured to determine the type of nitrogen source and carbon source of the medium and the respective corresponding contents according to the lipid content of the microorganism on the target band;
and the second determination submodule is used for determining a microbial spore drying method according to the protein content of the microorganism on the target wave band.
In summary, the scheme provided by the invention has the following technical effects:
according to the method, the particle size of the microorganism in a target wave band is determined to optimize the target, the liquid-solid ratio of the culture medium, the culture environment condition, the addition amount of inorganic salt and the spore inoculation amount on the culture medium, and the type and the content of a nitrogen source and a carbon source of the culture medium and a microbial spore drying method are determined according to the lipid content and the protein content of the microorganism in the target wave band, and the microorganism is cultured according to the particle size optimization target, the liquid-solid ratio of the culture medium, the culture environment condition, the addition amount of inorganic salt, the spore inoculation amount on the culture medium, the type and the content of the nitrogen source and the carbon source of the culture medium and the microbial spore drying method, so that the microorganism optical attenuation performance is improved and the broadband optical attenuation requirement is met on the premise of guaranteeing the spore yield of the microorganism; the invention realizes the controllability and operability of the optical attenuation performance of the microorganism by regulating and controlling the composition of the culture medium, the culture environment, the seed adding amount and the drying method, and further improves the application potential and development prospect of the microorganism material and the biological material in the field of optical attenuation.
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In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic flow chart of a method for controlling culture conditions for improving optical attenuation performance of microorganisms according to the present invention.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
This example shows a culture condition control method for improving the optical attenuation performance of microorganisms, and referring to fig. 1, the culture condition control method includes:
s1, acquiring a target wave band affecting the optical attenuation performance of microorganisms.
The target band in this embodiment is a common band affecting the optical attenuation performance of microorganisms, for example, the mid-infrared 3-5 μm band in BB spores is a common band affecting the optical attenuation performance of materials, and the 3-5 μm band is the target band.
S2, determining a particle size optimization target of the microorganism in the target wave band, a liquid-solid ratio of a culture medium, culture environment conditions, an inorganic salt addition amount and a spore inoculation amount on the culture medium.
In the embodiment, a discrete dipole approximation method is adopted, extinction sections of microorganisms under different particle sizes are calculated, the relations between the extinction sections and the particle sizes are compared, and the particle size corresponding to the maximum extinction section value is selected as the optimal particle size (particle size optimization target). The specific implementation process comprises the following steps:
and S21, acquiring microorganisms with different particle sizes.
The embodiment selectsNParticle size values of the individual particles are noted:r 1r 2r 3 ,…,r j ,…,r N
and S22, obtaining extinction sections corresponding to microorganisms with different particle sizes in the target wave band.
In the step, the specific process of obtaining the extinction section corresponding to the microorganisms with different particle sizes in the target wave band comprises the following steps:
step S221, respectively measuring the reflection spectrums of the microorganisms with different particle sizes in the target wave band to determine the reflection phase shift of the microorganisms with different particle sizes in the target wave band;
step S222, calculating complex refractive indexes corresponding to the microorganisms with different particle sizes in the target wave band according to the reflection spectrums and the corresponding reflection phase shifts of the microorganisms with different particle sizes in the target wave band;
the reflection spectrum of the microorganism is measured by using a Fourier infrared spectrometer, and the complex refractive index is obtained by adopting the Kramers-Kronig relation:
wherein,,m(lambda) is the complex refractive index;n(lambda) is complex refractive indexmThe real part of (lambda);k(lambda) is complex refractive indexmAn imaginary part of (λ);R(lambda) is the reflectance spectrum of the microorganism; Θ (λ) is the reflected phase shift, deduced from the reflection spectrum.
Step S223, according to the complex refractive index, calculating the dipole numbers corresponding to the microorganisms with different particle sizes in the target wave band and the dipole moment of each dipole;
the embodiment adoptsMThe microorganism is equivalent to a dipole,Mthe sum of the scattering intensities generated by the individual dipoles is equivalent to the scattering intensity of the microorganism. The present embodiment determines the dipole number using the following formula:
wherein,,Mis the dipole number; |m(lambda) is the complex refractive indexmA die of (lambda);r j is the firstjParticle size of individual particles.
In this embodiment, the dipole moment of each dipole is:
wherein,,P i is the firstiDipole moments of the individual dipoles;α i is the firstiThe polarizability of the individual dipoles;is the firstiIncidence of individual dipolesThe field strength; />To divide byiThe sum of the scattered field intensities generated by the remaining dipoles outside the individual dipoles;i=1,2,…,MMis the number of dipoles.
And step 224, calculating extinction sections corresponding to microorganisms with different particle sizes in the target wave band according to the dipole number and the dipole moment of each dipole.
This example uses an extinction cross-section to characterize the optical attenuation capabilities of microorganisms. The microbial extinction cross section of this example is:
wherein,,Cr j is of particle diameter ofr j Is a microbial extinction cross section;is the firstiIncident field electric field strength of individual dipoles +.>Complex conjugate of (a); lambda is the wavelength of the incident electromagnetic wave;j=1,2,…,NNis the particle size number of the particles.
S23, selecting the particle size corresponding to the maximum extinction section from extinction sections corresponding to microorganisms with different particle sizes in the target wave band as a particle size optimization target of the microorganisms in the target wave band;
and S24, judging the transformation trend between the optical attenuation of the microorganism and the particle size of the particles in the target wave band according to the extinction cross sections corresponding to the microorganisms with different particle sizes in the target wave band so as to determine the liquid-solid ratio of the culture medium, the culture environment condition, the addition amount of inorganic salt and the spore inoculation amount on the culture medium of the microorganisms.
The transformation trend in this embodiment is: the optical attenuation performance of the microorganism in a target band increases as the particle size of the microorganism particles increases; alternatively, the optical attenuation properties of the microorganism in the target wavelength band decrease as the particle size of the microorganism particles increases.
The selection of a suitable liquid-to-solid ratio is an important influencing factor affecting the growth of microorganisms. In the culture process, the excessive water content in the culture medium can inhibit the growth of microbial structures, and the porosity in the whole culture medium is reduced, so that the ventilation and the cooling in the culture medium are not facilitated. Too low a water content cannot meet the environmental requirements for microbial growth, affecting culture yield and particle morphology size. If the change trend is to increase the particle size of the aspergillus microorganism, the liquid-solid ratio of the culture medium is 2.5:1; if the trend of change is to reduce the particle size of the Aspergillus microorganism, the liquid-solid ratio of the culture medium is 1.5:1 or 3.5:1.
The culture environment conditions of this example include the culture environment temperature and the culture environment relative humidity. Since the vital activity of microorganisms is composed of a series of biochemical reactions, the reactions are extremely remarkably affected by temperature. Therefore, the temperature of the culture environment in the culture environment condition is also one of the most important factors affecting the growth of microorganisms. The intervention of the culture environment temperature on the growth of the microbial material mainly influences the mobility of microbial cell membranes and the activity of biological macromolecules, thereby influencing the life activities of the microorganisms. On one hand, as the ambient temperature increases, the enzyme reaction rate of the cell internal reference and the vital activity increases, and the metabolism and the growth of microorganisms correspondingly increase; on the other hand, with further elevation of temperature, active substances in microorganisms tend to denature, which causes the vital activities of cells to be affected. If the trend of change is to increase the particle size of the Aspergillus microorganism, the culture environment temperature is generally 26 ℃; if the trend is to reduce the particle size of the Aspergillus microorganism, the temperature of the culture environment is typically 22℃or 30 ℃.
The relative humidity of the culture environment in the culture environment condition is also a key parameter for culturing the microbial material, and relates to the growth of the thallus in the early stage and the sporulation in the later stage. However, the influence of the relative humidity in the environment on different stages of the cultivation of the microbial material is different, in the early growth stage, the relatively higher humidity can be beneficial to the growth of thalli, in the middle growth stage, the lower humidity is beneficial to the generation of spores, and in the final growth stage, the lower humidity can promote the end of the spore production process. If the change trend is to increase the particle size of the Aspergillus microorganism, the relative humidity of the culture environment in 1-4 days is 95-100%, the relative humidity of the culture environment in 5-6 days is 80%, and the relative humidity of the culture environment in 7 days is 60%; if the change trend is to reduce the particle size of the Aspergillus microorganism, the relative humidity of the culture environment in the 1 st to 4 th days is 95% -100%, and the relative humidity of the culture environment in the 5 th to 7 th days is 70%, so that the optimization target of smaller particle size can be realized while the normal yield stability of the microorganism is ensured.
The inorganic salt in the culture medium has very important significance for the growth of microbial materials, and has the main functions of participating in the cell composition during the growth of the microorganisms, and in addition, the content change of the inorganic salt can adjust the parameters such as osmotic pressure, pH value, oxidation-reduction potential and the like in the culture medium and can be used as an activator or inhibitor of enzymes to influence the growth of the organisms. If the trend of change is to increase the particle size of the Aspergillus microorganism, a higher amount of inorganic salt (KNO, for example) is selected 3 The addition amount was 0.3% MgSO 4 •7H 2 O addition 0.5%); if the trend of change is to reduce the particle size of the Aspergillus microorganism, a lower amount of inorganic salt (KNO 3 And MgSO 4 •7H 2 O added 0.1% each).
Whether the culture medium is a liquid culture medium or a solid culture medium, inoculating proper microbial mass on the culture medium plays a vital role in the growth of organisms, and when the inoculating biomass is too low, the nutrient growth time of the thalli is too long, so that the secretion and metabolism of enzymes are not facilitated; the inoculation spore quantity is too high, so that the growth among thalli can be greatly competitive, and the nutrition substances in the unit space are used up too early, so that the growth of organisms is not facilitated. If the trend is to increase the particle size of the Aspergillus microorganism, the volume of the medium (i.e., the spore inoculum size on the medium) is 0.3%; if the trend is to reduce the particle size of the Aspergillus microorganism, the volume of the medium (i.e., the spore inoculum size on the medium) is 0.9%.
S3, obtaining the lipid content and the protein content of the microorganism in the target wave band so as to determine the type and the content of a nitrogen source and a carbon source of the culture medium and a microorganism spore drying method.
The specific implementation process of the step comprises the following steps:
s31, setting constraint conditions of the content of each component in the microorganism;
the components in this example include proteins, lipids, nucleic acids and other trace elements. The constraint conditions are as follows:
wherein,,w p w l w n andw e the contents of protein, lipid, nucleic acid and other components respectively.
And S32, obtaining the average absorbance of each component on the target wave band so as to determine the average absorbance function of the microorganism on the target wave band.
The average absorbance function of the microorganism of this example over the target band is:
wherein,,for the microorganism in a target band lambda 12 Average absorbance thereon; />、/>And->Respectively proteins, nucleic acids, lipids and other components in the target band lambda 12 Average absorbance thereon;w p w n w l andw e the contents of proteins, nucleic acids, lipids and other constituents, respectively.
And step S33, constructing a composition optimization model according to the constraint conditions of the content of each composition component and the average absorbance function so as to determine the lipid content and the protein content of the microorganism on the target wave band.
The composition optimization model in this embodiment is:
and step S34, determining the types and the respective corresponding contents of the nitrogen source and the carbon source of the culture medium according to the lipid content of the microorganism on the target wave band.
The carbon source is not only a component constituting a carbon skeleton of the thallus, but also a substance energy source for the growth of the thallus. The addition of different carbon sources to the medium affects the composition of the microbial cell components. If the optimization objective is to obtain a higher lipid content, glucose or maltose should be selected as a carbon source, wherein if the lipid content is to be controlled further, the glucose addition can be adjusted and controlled, and the lipid content of the aspergillus microorganism is gradually increased when the glucose addition is changed from 2% to 12%, and if the optimization objective is to obtain a lower lipid content, glycerol, sucrose or lactose should be selected as a carbon source.
The nitrogen source in the culture medium directly influences the growth and the microorganismMetabolism. If the easy-consumption nitrogen source is adopted, hypha can grow rapidly within 24-48 hours without being controlled, so that the viscosity in the culture medium is increased, and the dissolved oxygen in the culture medium is rapidly reduced, which is unfavorable for further growth, development and metabolism of thalli. Therefore, the selection of nitrogen sources which are slowly metabolizable, inexpensive and have great significance for microbial cultivation. Adding inorganic nitrogen source such as (NH) into the culture medium 4 ) 2 SO 4 、NH 4 Cl、NaNO 3 、KNO 3 Or adding tryptone and urea as nitrogen source, microorganism hardly grows, if microorganism grows normally, yeast powder, peptone and corn steep liquor should be added as nitrogen source, if higher lipid content is to be obtained, corn steep liquor should be selected as nitrogen source, if moderate lipid content is to be obtained, peptone should be selected as nitrogen source, if lower lipid content is to be obtained, yeast powder should be selected as nitrogen source.
The method comprises the following steps: when the lipid content is 0-10%, the carbon source of the aspergillus microorganism culture medium is glycerol, sucrose or lactose, the adding amount is 3%, the nitrogen source is yeast powder, and the adding amount is 1%; when the lipid content is 10-20%, the carbon source of the aspergillus microorganism culture medium is maltose or glucose, the adding amount is 3%, the nitrogen source is yeast powder, and the adding amount is 1%; when the content is 20-30%, the carbon source of the aspergillus microorganism culture medium is glucose, the adding amount is 6%, the nitrogen source is yeast powder, and the adding amount is 2%. When the lipid content is more than 30%, the carbon of the aspergillus microorganism culture medium is glucose, the adding amount is 6%, the nitrogen source is peptone or corn steep liquor, and the adding amount is 2%.
And step S35, determining a microbial spore drying method according to the protein content of the microorganism on the target wave band.
When the protein content is less than 6%, the microbial spores in the fermentation broth are collected and dried by a drying method. When the protein content is less than 6%, the microbial spores in the fermentation broth are collected and dried by a lyophilization process.
S4, optimizing targets, a culture medium liquid-solid ratio, culture environment conditions, inorganic salt addition amount, spore inoculation amount on a culture medium, types and contents of a nitrogen source and a carbon source of the culture medium and a microorganism spore drying method according to the particle sizes of the particles, and culturing the microorganisms.
The optical attenuation performance of the microorganism is improved by controlling the culture condition of the microorganism.
According to the embodiment, the particle size of the microorganism in a target wave band is determined to optimize the target and the liquid-solid ratio of the culture medium, the culture environment condition, the addition amount of inorganic salt and the spore inoculation amount on the culture medium, and the type and the content of a nitrogen source and a carbon source of the culture medium and a microbial spore drying method are determined through the lipid content and the protein content of the microorganism in the target wave band, and the microorganism is cultured according to the particle size optimization target, the liquid-solid ratio of the culture medium, the culture environment condition, the addition amount of inorganic salt, the spore inoculation amount on the culture medium, the type and the content of the nitrogen source and the carbon source of the culture medium and the microbial spore drying method, so that the optical attenuation performance of the microorganism is improved on the premise of ensuring the spore yield and the metabolite yield of the microorganism, and the broadband optical attenuation requirement is met; the embodiment realizes the controllability and operability of the optical attenuation performance of the microorganism by regulating and controlling the composition of the culture medium, the culture environment, the seed adding amount and the drying method, and further improves the application potential and development prospect of the microorganism material and the biological material in the field of optical attenuation.
The above-described embodiments can be achieved by a culture condition control system for improving the optical attenuation performance of microorganisms as given in the following embodiments:
a culture condition control system for enhancing optical attenuation performance of microorganisms, the culture condition control system comprising:
the first acquisition module is used for acquiring a target wave band affecting the optical attenuation performance of the microorganism;
the determining module is used for determining a particle size optimization target of the microorganism in the target wave band, a liquid-solid ratio of a culture medium, culture environment conditions, an inorganic salt addition amount and a spore inoculation amount on the culture medium;
a second acquisition module for acquiring lipid content and protein content of the microorganism in the target wave band so as to determine the type and content of a nitrogen source and a carbon source of the culture medium and a microorganism spore drying method;
and the culture module is used for optimizing targets, a culture medium liquid-solid ratio, culture environment conditions, inorganic salt addition amount, spore inoculation amount on the culture medium, types and contents of a nitrogen source and a carbon source of the culture medium and a microbial spore drying method according to the particle size of the particles to culture the microorganisms.
Further, the determining module includes:
the first acquisition submodule is used for acquiring microorganisms with different particle sizes;
the second acquisition submodule is used for acquiring extinction sections corresponding to microorganisms with different particle sizes in the target wave band;
selecting a submodule, namely selecting the particle size corresponding to the maximum extinction cross section from extinction cross sections corresponding to microorganisms with different particle sizes in the target wave band as a particle size optimization target of the microorganisms in the target wave band;
the judging submodule is used for judging the transformation trend between the optical attenuation of the microorganism in the target wave band and the particle size of the particles according to the extinction cross sections corresponding to the microorganisms with different particle sizes in the target wave band so as to determine the liquid-solid ratio of the culture medium of the microorganisms, the culture environment condition, the addition amount of inorganic salt and the spore inoculation amount on the culture medium;
the transformation trend is: the microbial optical attenuation performance increases as the particle size of the microbial particles increases; alternatively, the microbial optical attenuation performance decreases as the microbial particle size increases.
Further, the second obtaining submodule includes:
the measuring submodule is used for respectively measuring the reflection spectrums of the microorganisms with different particle sizes in the target wave band so as to determine the reflection phase shift of the microorganisms with different particle sizes in the target wave band;
the first calculating submodule is used for calculating complex refractive indexes corresponding to the microorganisms with different particle sizes in the target wave band according to the reflection spectrums and the corresponding reflection phase shifts of the microorganisms with different particle sizes in the target wave band;
the second calculation submodule is used for calculating the dipole number and the dipole moment of each dipole corresponding to the microorganisms with different particle sizes in the target wave band according to the complex refractive index;
and the third calculation sub-module is used for calculating extinction sections corresponding to microorganisms with different particle sizes in the target wave band according to the dipole number and the dipole moment of each dipole.
Further, the second obtaining module includes:
the setting submodule is used for setting constraint conditions of the content of each component in the microorganism;
the components comprise proteins, lipids, nucleic acids and other microelements;
a third obtaining submodule, configured to obtain average absorbance of each component on the target band, so as to determine an average absorbance function of the microorganism on the target band;
the constructing submodule is used for constructing a composition optimizing model according to the constraint conditions of the content of each composition component and the average absorbance function so as to determine the lipid content and the protein content of the microorganism on the target wave band;
a first determining submodule, configured to determine the type of nitrogen source and carbon source of the medium and the respective corresponding contents according to the lipid content of the microorganism on the target band;
and the second determination submodule is used for determining a microbial spore drying method according to the protein content of the microorganism on the target wave band.
The principles, formulas and parameter definitions according to the above embodiments are applicable, and will not be traced back.
Note that the technical features of the above embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be regarded as the scope of the description. The foregoing examples represent only a few embodiments of the present application, which are described in more detail and are not to be construed as limiting the scope of the invention. It should be noted that it would be apparent to those skilled in the art that various modifications and improvements could be made without departing from the spirit of the present application, which would be within the scope of the present application. Accordingly, the scope of protection of the present application is to be determined by the claims appended hereto.

Claims (4)

1.一种提升微生物光学衰减性能的培养条件控制方法,其特征在于,所述培养条件控制方法包括:1. A culture condition control method that promotes microbial optical attenuation performance, it is characterized in that, described culture condition control method comprises: 步骤S1、获取影响微生物光学衰减性能的目标波段;Step S1, obtaining the target waveband that affects the optical attenuation performance of microorganisms; 步骤S2、确定所述微生物在所述目标波段内的颗粒粒径优化目标、培养基液固比、培养环境条件、无机盐添加量和培养基上孢子接种量;Step S2, determining the particle size optimization target of the microorganism in the target band, the liquid-solid ratio of the culture medium, the culture environment conditions, the amount of inorganic salt added, and the amount of spore inoculum on the culture medium; 步骤S3、获取所述微生物在所述目标波段内的脂质含量和蛋白质含量,以确定培养基氮源和碳源的种类和含量以及微生物孢子干燥方法;Step S3, obtaining the lipid content and protein content of the microorganism in the target band, so as to determine the type and content of the nitrogen source and carbon source of the medium and the drying method of the microbial spores; 步骤S4、按照所述颗粒粒径优化目标、培养基液固比、培养环境条件、无机盐添加量、培养基上孢子接种量、培养基氮源和碳源的种类和含量以及微生物孢子干燥方法,培养所述微生物;Step S4, optimize the target according to the particle size, the liquid-solid ratio of the culture medium, the culture environment conditions, the amount of inorganic salt added, the amount of spore inoculum on the medium, the type and content of the nitrogen source and carbon source of the medium, and the drying method of microbial spores , cultivating the microorganism; 所述步骤S2的具体实现过程包括:The concrete realization process of described step S2 comprises: 步骤S21、获取不同颗粒粒径的微生物;Step S21, obtaining microorganisms with different particle sizes; 步骤S22、获取所述目标波段内不同颗粒粒径的微生物对应的消光截面;Step S22, obtaining the extinction cross section corresponding to microorganisms with different particle sizes in the target band; 步骤S23、从所述目标波段内不同颗粒粒径的微生物对应的消光截面中选取出最大消光截面对应的颗粒粒径作为所述微生物在所述目标波段内的颗粒粒径优化目标;Step S23, selecting the particle size corresponding to the largest extinction cross section from the extinction cross sections corresponding to microorganisms with different particle sizes in the target band as the particle size optimization target of the microorganisms in the target band; 步骤S24、根据所述目标波段内不同颗粒粒径的微生物对应的消光截面,对所述目标波段内的所述微生物光学衰减性和颗粒粒径之间的变换趋势进行判断,以确定所述微生物的培养基液固比、培养环境条件、无机盐添加量和培养基上孢子接种量;Step S24, according to the extinction cross section corresponding to microorganisms with different particle sizes in the target waveband, judge the transformation trend between the optical attenuation of the microorganisms in the target waveband and the particle size, so as to determine the microbial The liquid-solid ratio of the medium, the culture environment conditions, the amount of inorganic salt added and the amount of spore inoculation on the medium; 所述变换趋势为:所述微生物光学衰减性能随所述微生物颗粒粒径增大而增大;或者,所述微生物光学衰减性能随所述微生物颗粒粒径增大而减小;The conversion trend is: the optical attenuation performance of the microorganisms increases with the particle size of the microorganism particles; or, the optical attenuation performance of the microorganisms decreases with the particle size of the microorganism particles; 在所述步骤S22中,所述步骤S22中,所述获取所述目标波段内不同颗粒粒径的微生物对应的消光截面的具体过程包括:In the step S22, in the step S22, the specific process of obtaining the corresponding extinction cross section of microorganisms with different particle sizes in the target band includes: 步骤S221、对所述目标波段内不同颗粒粒径的微生物的反射谱分别进行测量,以确定所述目标波段内不同颗粒粒径的微生物的反射相移;Step S221: Measure the reflectance spectra of microorganisms with different particle sizes in the target band, respectively, to determine the reflection phase shift of microorganisms with different particle sizes in the target band; 步骤S222、根据所述目标波段内不同颗粒粒径的微生物的反射谱和对应的反射相移,计算所述目标波段内不同颗粒粒径的微生物各自对应的复折射率;Step S222, according to the reflectance spectra and corresponding reflection phase shifts of microorganisms with different particle sizes in the target band, calculate the complex refractive indices corresponding to the microorganisms with different particle sizes in the target band; 步骤S223、根据所述复折射率,计算所述目标波段内不同颗粒粒径的微生物各自对应的偶极子数以及每个偶极子的偶极矩;Step S223, according to the complex refractive index, calculate the number of dipoles corresponding to microorganisms with different particle sizes in the target band and the dipole moment of each dipole; 步骤S224、根据所述偶极子数以及每个偶极子的偶极矩,计算所述目标波段内不同颗粒粒径的微生物对应的消光截面;Step S224, according to the number of dipoles and the dipole moment of each dipole, calculate the extinction cross section corresponding to microorganisms with different particle sizes in the target band; 在所述步骤S223中,所述每个偶极子的偶极矩为:In the step S223, the dipole moment of each dipole is: P i =α i (E inc , i +E sca , i ); P i = α i ( E inc , i + E sca , i ); 其中,P i 为第i个偶极子的偶极矩;α i 为第i个偶极子的极化率;E inc , i 为第i个偶极子的入射场电场强度;E sca , i 为除第i个偶极子外剩余偶极子所产生的散射场强度之和;i=1,2,…,MM为偶极子数;Among them, P i is the dipole moment of the i-th dipole; α i is the polarizability of the i -th dipole; E inc , i is the electric field intensity of the i- th dipole; E sca , i is the sum of the scattered field intensity produced by the remaining dipoles except the i -th dipole; i =1, 2,..., M , M is the number of dipoles; 在所述步骤S224中,所述微生物消光截面为:In the step S224, the microbial extinction cross section is: ; 其中,Cr j 为颗粒粒径为r j 的微生物消光截面;为第i个偶极子的入射场电场强度E inc , i 的复共轭;/>为入射电磁波的波长;j=1,2,…,NN为颗粒粒径数。Wherein, Cr j is the microbial extinction cross section whose particle size is r j ; is the incident field electric field intensity E inc of the i- th dipole, the complex conjugate of i ; /> is the wavelength of the incident electromagnetic wave; j =1, 2,..., N , N is the particle size number. 2.根据权利要求1所述的培养条件控制方法,其特征在于,所述步骤S3的具体实现过程包括:2. The culture condition control method according to claim 1, characterized in that, the specific implementation process of the step S3 comprises: 步骤S31、设置所述微生物中各个组成成分含量的约束条件;Step S31, setting constraints on the content of each component in the microorganism; 所述组成成分包括蛋白质、脂质、核酸和其他微量元素;The composition includes proteins, lipids, nucleic acids and other trace elements; 步骤S32、获取所述各个组成成分在所述目标波段上各自对应的平均吸光度,以确定所述微生物在所述目标波段上的平均吸光度函数;Step S32, obtaining the respective average absorbance of each component on the target band, so as to determine the average absorbance function of the microorganism on the target band; 步骤S33、根据所述各个组成成分含量的约束条件以及所述平均吸光度函数,构建组成成分优化模型,以确定所述微生物在所述目标波段上的脂质含量和蛋白质含量;Step S33, according to the constraints of the content of each component and the average absorbance function, construct a component optimization model to determine the lipid content and protein content of the microorganism in the target band; 步骤S34、根据所述微生物在所述目标波段上的脂质含量,确定培养基氮源和碳源的种类以及各自对应的含量;Step S34, according to the lipid content of the microorganism in the target band, determine the types of nitrogen source and carbon source in the medium and their corresponding contents; 步骤S35、根据所述微生物在所述目标波段上的蛋白质含量,确定微生物孢子干燥方法。Step S35 , according to the protein content of the microorganism in the target band, determine the drying method of the microorganism spores. 3.一种提升微生物光学衰减性能的培养条件控制系统,其特征在于,所述培养条件控制系统包括:3. a culture condition control system that promotes microbial optical attenuation performance, it is characterized in that, described culture condition control system comprises: 第一获取模块,用于获取影响微生物光学衰减性能的目标波段;The first acquisition module is used to acquire the target waveband that affects the optical attenuation performance of microorganisms; 确定模块,用于确定所述微生物在所述目标波段内的颗粒粒径优化目标、培养基液固比、培养环境条件、无机盐添加量和培养基上孢子接种量;A determination module, configured to determine the particle size optimization target of the microorganism in the target wave band, the liquid-solid ratio of the culture medium, the culture environment conditions, the amount of inorganic salt added and the amount of spore inoculum on the culture medium; 第二获取模块,用于获取所述微生物在所述目标波段内的脂质含量和蛋白质含量,以确定培养基氮源和碳源的种类和含量以及微生物孢子干燥方法;The second acquisition module is used to acquire the lipid content and protein content of the microorganism in the target band, so as to determine the type and content of the nitrogen source and carbon source of the culture medium and the drying method of the microbial spores; 培养模块,用于按照所述颗粒粒径优化目标、培养基液固比、培养环境条件、无机盐添加量、培养基上孢子接种量、培养基氮源和碳源的种类和含量以及微生物孢子干燥方法,培养所述微生物;The cultivation module is used to optimize the target according to the particle size, the liquid-solid ratio of the culture medium, the culture environment conditions, the amount of inorganic salt added, the amount of spore inoculum on the medium, the type and content of the nitrogen source and carbon source of the medium, and microbial spores drying method, cultivating said microorganism; 所述确定模块包括:The determination module includes: 第一获取子模块,用于获取不同颗粒粒径的微生物;The first acquisition sub-module is used to acquire microorganisms with different particle sizes; 第二获取子模块,用于获取所述目标波段内不同颗粒粒径的微生物对应的消光截面;The second acquisition sub-module is used to acquire the extinction cross section corresponding to microorganisms with different particle sizes in the target band; 选取子模块,用于从所述目标波段内不同颗粒粒径的微生物对应的消光截面中选取出最大消光截面对应的颗粒粒径作为所述微生物在所述目标波段内的颗粒粒径优化目标;Selecting a sub-module for selecting the particle size corresponding to the largest extinction cross section from the extinction cross sections corresponding to microorganisms with different particle sizes in the target band as the particle size optimization target of the microorganisms in the target band; 判断子模块,用于根据所述目标波段内不同颗粒粒径的微生物对应的消光截面,对所述目标波段内的所述微生物光学衰减性和颗粒粒径之间的变换趋势进行判断,以确定所述微生物的培养基液固比、培养环境条件、无机盐添加量和培养基上孢子接种量;The judging sub-module is used to judge the transformation trend between the microbial optical attenuation and the particle size in the target band according to the extinction cross section corresponding to the microorganisms with different particle sizes in the target band, so as to determine The liquid-solid ratio of the culture medium, the culture environment conditions, the amount of inorganic salt added and the spore inoculum amount on the culture medium of the microorganism; 所述变换趋势为:所述微生物光学衰减性能随所述微生物颗粒粒径增大而增大;或者,所述微生物光学衰减性能随所述微生物颗粒粒径增大而减小;The conversion trend is: the optical attenuation performance of the microorganisms increases with the particle size of the microorganism particles; or, the optical attenuation performance of the microorganisms decreases with the particle size of the microorganism particles; 所述第二获取子模块包括:The second acquisition submodule includes: 测量子模块,用于对所述目标波段内不同颗粒粒径的微生物的反射谱分别进行测量,以确定所述目标波段内不同颗粒粒径的微生物的反射相移;The measurement sub-module is used to measure the reflection spectra of microorganisms with different particle sizes in the target band, respectively, so as to determine the reflection phase shift of microorganisms with different particle sizes in the target band; 第一计算子模块,用于根据所述目标波段内不同颗粒粒径的微生物的反射谱和对应的反射相移,计算所述目标波段内不同颗粒粒径的微生物各自对应的复折射率;The first calculation sub-module is used to calculate the corresponding complex refractive indices of microorganisms with different particle sizes in the target wave band according to the reflection spectra and corresponding reflection phase shifts of microorganisms with different particle sizes in the target wave band; 第二计算子模块,用于根据所述复折射率,计算所述目标波段内不同颗粒粒径的微生物各自对应的偶极子数以及每个偶极子的偶极矩;The second calculation submodule is used to calculate the number of dipoles corresponding to microorganisms with different particle sizes in the target band and the dipole moment of each dipole according to the complex refractive index; 第三计算子模块,用于根据所述偶极子数以及每个偶极子的偶极矩,计算所述目标波段内不同颗粒粒径的微生物对应的消光截面;The third calculation submodule is used to calculate the corresponding extinction cross section of microorganisms with different particle sizes in the target band according to the number of dipoles and the dipole moment of each dipole; 所述每个偶极子的偶极矩为:The dipole moment of each dipole is: P i =α i (E inc , i +E sca , i ); P i = α i ( E inc , i + E sca , i ); 其中,P i 为第i个偶极子的偶极矩;α i 为第i个偶极子的极化率;E inc , i 为第i个偶极子的入射场电场强度;E sca , i 为除第i个偶极子外剩余偶极子所产生的散射场强度之和;i=1,2,…,MM为偶极子数;Among them, P i is the dipole moment of the i-th dipole; α i is the polarizability of the i -th dipole; E inc , i is the electric field intensity of the i- th dipole; E sca , i is the sum of the scattered field intensity produced by the remaining dipoles except the i -th dipole; i =1, 2,..., M , M is the number of dipoles; 所述微生物消光截面为:The microbial extinction cross section is: ; 其中,Cr j 为颗粒粒径为r j 的微生物消光截面;为第i个偶极子的入射场电场强度E inc , i 的复共轭;/>为入射电磁波的波长;j=1,2,…,NN为颗粒粒径数。Wherein, Cr j is the microbial extinction cross section whose particle size is r j ; is the incident field electric field intensity E inc of the i- th dipole, the complex conjugate of i ; /> is the wavelength of the incident electromagnetic wave; j =1, 2,..., N , N is the particle size number. 4.根据权利要求3所述的培养条件控制系统,其特征在于,所述第二获取模块包括:4. The culture condition control system according to claim 3, wherein the second acquisition module comprises: 设置子模块,用于设置所述微生物中各个组成成分含量的约束条件;Setting submodules, used to set constraints on the content of each component in the microorganisms; 所述组成成分包括蛋白质、脂质、核酸和其他微量元素;The composition includes proteins, lipids, nucleic acids and other trace elements; 第三获取子模块,用于获取所述各个组成成分在所述目标波段上各自对应的平均吸光度,以确定所述微生物在所述目标波段上的平均吸光度函数;The third acquisition sub-module is used to acquire the respective average absorbance of each component on the target wave band, so as to determine the average absorbance function of the microorganism on the target wave band; 构建子模块,用于根据所述各个组成成分含量的约束条件以及所述平均吸光度函数,构建组成成分优化模型,以确定所述微生物在所述目标波段上的脂质含量和蛋白质含量;Constructing submodules for constructing a component optimization model according to the constraint conditions of the content of each component and the average absorbance function, so as to determine the lipid content and protein content of the microorganism in the target band; 第一确定子模块,用于根据所述微生物在所述目标波段上的脂质含量,确定培养基氮源和碳源的种类以及各自对应的含量;The first determination submodule is used to determine the types of nitrogen source and carbon source of the medium and their corresponding contents according to the lipid content of the microorganism on the target band; 第二确定子模块,用于根据所述微生物在所述目标波段上的蛋白质含量,确定微生物孢子干燥方法。The second determination sub-module is used to determine the drying method of microbial spores according to the protein content of the microorganisms in the target band.
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