Method and system for extracting natural pigment for coating based on microbial fermentation

Abstract

The invention relates to the technical field of material chemical industry, in particular to a method and a system for extracting natural pigment for coating based on microbial fermentation, which comprises the following steps: acquiring a plurality of initial strains, acquiring a genetic recombination strain group, preparing single-cell suspension from the genetic recombination strains, performing fluorescence detection on the single-cell suspension, and confirming an optimal mutant strain. The method comprises the following steps: obtaining a mutant strain pigment crude extract, confirming a maximum absorption wavelength, culturing an optimal mutant strain, respectively carrying out absorbance determination on each mutant strain experimental group to obtain an absorbance value set, confirming an optimal fermentation temperature, carrying out large-scale fermentation to obtain a high-secretion fermentation liquid, carrying out wall breaking treatment to obtain a wall-broken thallus suspension, and carrying out freeze drying to obtain the high-secretion thallus suspension. The method comprises the following steps: preparing an intracellular pigment solution, carrying out microencapsulation on the intracellular pigment solution to obtain pigment microspheres, extracting a verification pigment extracting solution from the pigment microspheres, and obtaining the natural pigment by utilizing the verification pigment extracting solution. The problems of low extraction efficiency, high energy consumption and environmental pollution during extraction of the natural pigment for the coating can be solved.

Description

Technical Field

[0001] This invention relates to the field of materials and chemical engineering, and in particular to a method and system for extracting natural pigments for coatings based on microbial fermentation. Background Technology

[0002] With the rapid development of the materials and chemical industry, the extraction of natural pigments for coatings faces new challenges. Gene editing technology can be used to optimize bacterial strains, improving pigment yield and quality, thereby addressing the problems of low extraction efficiency, high energy consumption, and environmental pollution associated with the extraction of natural pigments for coatings.

[0003] Currently, natural pigments are mainly extracted using traditional organic solvents. While traditional methods can yield natural pigments, they suffer from low extraction efficiency, high energy consumption, and environmental pollution. Therefore, optimizing the extraction of natural pigments for coatings is crucial for improving extraction efficiency. Summary of the Invention

[0004] This invention provides a method for extracting natural pigments for coatings based on microbial fermentation and a computer-readable storage medium. Its main purpose is to solve the problems of low extraction efficiency, high energy consumption and environmental pollution in the extraction of natural pigments for coatings.

[0005] To achieve the above objectives, the present invention provides a method for extracting natural pigments for coatings based on microbial fermentation, comprising:

[0006] Multiple initial strains were obtained, and a group of recombinant strains was obtained using the multiple initial strains and pre-constructed gene editing technology. The group of recombinant strains included multiple recombinant strains.

[0007] For each of the recombinant strains in the aforementioned recombinant strain group, a single-cell suspension was prepared to obtain a single-cell suspension set. The single-cell suspension set included multiple single-cell suspensions, and each single-cell suspension corresponded one-to-one with a recombinant strain. Multiple single-cell suspensions were subjected to fluorescence detection to obtain multiple fluorescence detection nodes. The optimal mutant strain was identified using multiple fluorescence detection nodes.

[0008] The optimal mutant strain was subjected to crude pigment extraction to obtain a crude extract of mutant pigment. The maximum absorption wavelength was determined using the crude extract of mutant pigment.

[0009] The best mutant strains were cultivated to obtain multiple best mutant strains. The multiple best mutant strains were grouped according to the preset fermentation temperature to obtain multiple mutant strain experimental groups. The absorbance of each mutant strain experimental group in the multiple mutant strain experimental groups was measured to obtain an absorbance value set. The optimal fermentation temperature was determined using the absorbance value set. The absorbance value set includes the absorbance value of each mutant strain experimental group in the multiple mutant strain experimental groups at the maximum absorption wavelength.

[0010] High-secretion fermentation broth was obtained by using the optimal fermentation temperature and the optimal mutant strain for large-scale fermentation.

[0011] The high-secretion fermentation broth was subjected to cell wall disruption to obtain a cell wall disruption suspension. A dual-enzyme synergistic pigment release operation was performed on the cell wall disruption suspension to obtain an intracellular pigment solution. The intracellular pigment solution was then microencapsulated to obtain pigment microspheres.

[0012] A verification pigment extract was extracted from the pigment microspheres. After confirming that the verification pigment extract was the preset target pigment extract, the natural pigment was obtained using the verification pigment extract.

[0013] Optionally, obtaining multiple initial strains includes:

[0014] Multiple candidate strains were obtained using a pre-constructed marine microbial database;

[0015] Gene sequence alignment analysis was performed on all the candidate strains, resulting in multiple classification and attribution reports;

[0016] Based on the multiple classification and attribution reports, multiple target strains were identified from multiple candidate strains;

[0017] Each of the multiple target strains was cultured in pre-confirmed dried seawater medium to obtain multiple stable target strains, with each target strain corresponding to a stable target strain.

[0018] Intracellular pigment content was detected for each of the multiple stable target strains to obtain a set of intracellular pigment content values. The set of intracellular pigment content values ​​included multiple intracellular pigment content values, and each intracellular pigment content value corresponded one-to-one with a stable target strain.

[0019] The stable target strains corresponding to intracellular pigment content values ​​that are higher than the preset intracellular pigment content threshold are identified as initial strains, resulting in multiple initial strains.

[0020] Optionally, the method of obtaining a gene recombinant strain population using multiple initial strains and pre-constructed gene editing technology includes:

[0021] For each of the multiple initial strains, the following procedure was performed:

[0022] The initial strain was physically isolated to obtain experimental strains and control strains;

[0023] Whole-genome sequencing was performed on the experimental strain to obtain its genomic information;

[0024] The experimental strain was recombined using its genomic information and pre-constructed gene editing technology to obtain a primary gene-recombined strain.

[0025] The primary gene recombinant strain and the control strain were fermented separately to obtain recombinant fermentation cell liquid and control fermentation cell liquid, respectively.

[0026] The recombinant fermentation cell broth and the control fermentation cell broth were subjected to crude pigment extraction to obtain crude recombinant pigment extract and crude control pigment extract, respectively.

[0027] Obtain the pigment mass concentration of the crude extract of recombinant pigment and the pigment mass concentration of the crude extract of control pigment;

[0028] The yield increase rate was calculated using the pigment concentration of the recombinant pigment crude extract and the pigment concentration of the control pigment crude extract.

[0029] The yield increase rate is compared with the preset yield increase threshold, and the primary gene recombinant strains corresponding to the yield increase rate that is greater than or equal to the yield increase threshold are identified as gene recombinant strains.

[0030] Multiple recombinant strains were combined to obtain a group of recombinant strains.

[0031] Optionally, the fluorescence detection of multiple single-cell suspensions to obtain multiple fluorescence detection nodes includes:

[0032] Perform the following operation on each of the multiple single-cell suspensions:

[0033] The single-cell suspension was introduced into a pre-constructed nanoreactor to obtain an initial nanoreactor;

[0034] A pre-confirmed fluorescent reporter gene expression vector was added to the initial nanoreactor to obtain the labeled nanoreactor;

[0035] The labeled nanoreactors were placed in a pre-confirmed constant temperature environment and cultured for 48 hours to obtain labeled microculture medium. Every 4 hours within 48 hours, the cell fluorescence density of the labeled nanoreactors was detected using a pre-confirmed high-throughput fluorescence microscope to obtain cell density sequences and fluorescence intensity sequences.

[0036] Cell density-time curves were constructed using cell density sequences, and the maximum specific growth rate was obtained based on the cell density-time curves.

[0037] The maximum pigment synthesis activity index was obtained based on cell density and fluorescence intensity sequences.

[0038] The coefficient of variation was calculated from the fluorescence intensity sequence to obtain the coefficient of variation.

[0039] The maximum specific growth rate, maximum pigment synthesis activity index, and coefficient of variation are summarized to obtain the fluorescence detection node;

[0040] By summarizing the fluorescence detection nodes, multiple fluorescence detection nodes are obtained, and each fluorescence detection node corresponds one-to-one with a labeled microculture medium.

[0041] Optionally, the step of identifying the optimal mutant strain using multiple fluorescence detection nodes includes:

[0042] The maximum specific growth rate, maximum pigment synthesis activity index, and coefficient of variation in multiple fluorescence detection nodes were normalized to obtain multiple normalized fluorescence detection nodes. The normalized fluorescence detection nodes include the normalized maximum specific growth rate, the normalized maximum pigment synthesis activity index, and the normalized coefficient of variation.

[0043] A comprehensive evaluation value is calculated using each of the multiple normalized fluorescence detection nodes and a pre-constructed comprehensive evaluation value calculation formula to obtain a comprehensive evaluation value set. The comprehensive evaluation value calculation formula is as follows:

[0044] S = 0.4 × T max +0.5×L+0.1×(1-CV)

[0045] Where S represents the comprehensive evaluation value, T max denoted as the normalized maximum specific growth rate, L represents the normalized maximum pigment synthesis activity index, and CV represents the normalized coefficient of variation.

[0046] The comprehensive evaluation values ​​in the comprehensive evaluation value set are arranged in descending order to obtain a comprehensive evaluation value sequence. The top 5 comprehensive evaluation values ​​are extracted from the comprehensive evaluation value sequence. The labeled micro-culture media corresponding to the extracted 5 comprehensive evaluation values ​​are all identified as candidate micro-culture media, thus obtaining 5 candidate culture media.

[0047] Verification experiments were conducted on each of the five candidate culture media, resulting in five experimental reports. Based on these reports, the optimal labeled microculture medium was determined, and the gene recombinant strain corresponding to the optimal labeled microculture medium was identified as the optimal mutant strain.

[0048] Optionally, the step of crudely extracting pigments from the optimal mutant strain to obtain a crude extract of mutant pigments, and using the crude extract of mutant pigments to determine the maximum absorption wavelength, includes:

[0049] The optimal mutant strain was fermented to obtain the mutant strain fermentation broth, and the mutant strain fermentation broth was centrifuged to obtain cell precipitate;

[0050] The bacterial cell precipitate was resuspended to obtain a bacterial cell suspension, and the bacterial cell suspension was then broken down to obtain a cell lysate.

[0051] The cell lysate was subjected to static extraction to obtain an extraction mixture. The extraction mixture was then centrifuged to obtain a supernatant, which was used as the crude extract of pigments from the mutant strain.

[0052] Absorption spectral data were obtained by scanning the crude pigment extract of the mutant strain using a pre-confirmed UV-Vis spectrophotometer across the entire wavelength range.

[0053] Peak identification is performed on the absorption spectrum data to obtain the maximum absorption wavelength.

[0054] Optionally, the absorbance measurement is performed on each of the multiple mutant experimental groups to obtain an absorbance value set, and the optimal fermentation temperature is determined using the absorbance value set, including:

[0055] Each mutant experimental group in the multiple mutant experimental groups was inoculated into a pre-confirmed fermentation medium and cultured to obtain a fermentation broth sample set, wherein the fermentation broth sample set includes multiple fermentation broth samples, and the fermentation broth samples correspond one-to-one with the mutant experimental groups.

[0056] Multiple fermentation broth samples were pretreated to obtain a supernatant sample set, which included multiple supernatant samples.

[0057] The absorbance of multiple supernatant samples was measured using the maximum absorption wavelength and a UV-Vis spectrophotometer to obtain a set of absorbance values.

[0058] Construct a temperature-absorbance curve using the absorbance value set;

[0059] The optimal fermentation temperature was determined using the temperature-absorbance curve.

[0060] Optionally, the step of microencapsulating the intracellular pigment fluid to obtain pigment microspheres includes:

[0061] The intracellular pigment solution and the pre-confirmed aerogel-sodium alginate composite emulsion were homogenized and emulsified at a preset homogenization speed to obtain a primary emulsion.

[0062] The primary emulsion was ion-gelled to obtain multiple primary microspheres;

[0063] Each primary microsphere in a plurality of primary microspheres was cross-linked and embedded in a pre-confirmed chitosan buffer to obtain a plurality of double-embedded wet microspheres;

[0064] Multiple double-embedded wet microspheres were freeze-dried under vacuum to obtain multiple original pigment microspheres;

[0065] Random sampling was performed on multiple original pigment microspheres to obtain a sampled original pigment microsphere set and a set of undetermined original pigment microspheres. The sampled original pigment microsphere set included multiple sampled original pigment microspheres, and the set of undetermined original pigment microspheres included multiple undetermined original pigment microspheres.

[0066] The pigment embedding rate of multiple sampled original pigment microspheres was detected using a pre-confirmed pigment embedding rate detector, resulting in a set of pigment embedding rate detection values, which included multiple pigment embedding rate detection values.

[0067] Multiple pigment embedding rate detection values ​​are compared with preset pigment embedding rate detection thresholds. Sampled original pigment microspheres with pigment embedding rate detection values ​​greater than or equal to the pigment embedding rate detection thresholds are identified as qualified original pigment microspheres. The qualified original pigment microspheres and sampled original pigment microspheres are used to obtain the original pigment microsphere qualification rate.

[0068] The pass rate of the original pigment microspheres is compared with the preset pass rate threshold of the original pigment microspheres. If the pass rate of the original pigment microspheres is less than the threshold, the homogenization speed is adjusted and the adjusted homogenization speed is used as the preset homogenization speed. The process is repeated until the pass rate of the original pigment microspheres is greater than or equal to the threshold. The multiple undetermined original pigment microspheres are then confirmed as pigment microspheres.

[0069] Optionally, after confirming that the verification pigment extract is the preset target pigment extract, the natural pigment is obtained using the verification pigment extract, including:

[0070] The pigment extract was mixed with the pre-confirmed acrylic resin emulsion to obtain the paint to be tested.

[0071] The pre-confirmed aluminum plate was sprayed with the color paint to be tested to obtain the sprayed aluminum plate.

[0072] Color difference detection is performed on the coated aluminum plate to obtain the color difference detection value. The color difference detection value is compared with the preset color difference detection threshold. If the color difference detection value is less than the color difference detection threshold, the verification pigment extract is confirmed as the target pigment extract.

[0073] The target pigment extract was purified to obtain the natural pigment.

[0074] To achieve the above objectives, the present invention also provides a natural pigment extraction system for coatings based on microbial fermentation, comprising:

[0075] The mutant strain screening module is used to obtain multiple initial strains and to obtain a gene recombinant strain population using multiple initial strains and pre-constructed gene editing technology. The gene recombinant strain population includes multiple gene recombinant strains.

[0076] For each of the recombinant strains in the aforementioned recombinant strain group, a single-cell suspension was prepared to obtain a single-cell suspension set. The single-cell suspension set included multiple single-cell suspensions, and each single-cell suspension corresponded one-to-one with a recombinant strain. Multiple single-cell suspensions were subjected to fluorescence detection to obtain multiple fluorescence detection nodes. The optimal mutant strain was identified using multiple fluorescence detection nodes.

[0077] The fermentation condition confirmation module is used to perform crude pigment extraction on the optimal mutant strain to obtain a crude pigment extract of the mutant strain, and to confirm the maximum absorption wavelength using the crude pigment extract of the mutant strain.

[0078] The best mutant strains were cultivated to obtain multiple best mutant strains. The multiple best mutant strains were grouped according to the preset fermentation temperature to obtain multiple mutant strain experimental groups. The absorbance of each mutant strain experimental group in the multiple mutant strain experimental groups was measured to obtain an absorbance value set. The optimal fermentation temperature was determined using the absorbance value set. The absorbance value set includes the absorbance value of each mutant strain experimental group in the multiple mutant strain experimental groups at the maximum absorption wavelength.

[0079] The pigment microsphere synthesis module is used to carry out large-scale fermentation using the optimal fermentation temperature and the optimal mutant strain to obtain a high-secretion fermentation broth;

[0080] The high-secretion fermentation broth was subjected to cell wall disruption to obtain a cell wall disruption suspension. A dual-enzyme synergistic pigment release operation was performed on the cell wall disruption suspension to obtain an intracellular pigment solution. The intracellular pigment solution was then microencapsulated to obtain pigment microspheres.

[0081] The natural pigment verification module is used to extract a verification pigment extract from the pigment microspheres. After confirming that the verification pigment extract is the preset target pigment extract, the natural pigment is obtained using the verification pigment extract.

[0082] To address the above problems, the present invention also provides an electronic device, the electronic device comprising:

[0083] Memory, storing at least one instruction; and

[0084] The processor executes the instructions stored in the memory to implement the above-described method for extracting natural pigments for coatings based on microbial fermentation.

[0085] To address the aforementioned problems, the present invention also provides a computer-readable storage medium storing at least one instruction, which is executed by a processor in an electronic device to implement the above-described method for extracting natural pigments for coatings based on microbial fermentation.

[0086] To address the problems described in the background art, this invention obtains multiple initial bacterial strains and utilizes pre-constructed gene editing technology to obtain a recombinant bacterial strain population. The recombinant bacterial strain population includes multiple recombinant bacterial strains. For each recombinant bacterial strain in the population, single-cell suspensions are prepared to obtain a set of single-cell suspensions. Each single-cell suspension set includes multiple single-cell suspensions, and each single-cell suspension corresponds one-to-one with a recombinant bacterial strain. Fluorescence detection is performed on multiple single-cell suspensions to obtain multiple fluorescence detection nodes. The optimal mutant strain is identified using these multiple fluorescence detection nodes. Therefore, this invention, by obtaining multiple initial bacterial strains, constructing a recombinant bacterial strain population using gene editing technology, and then screening for the optimal mutant strain through fluorescence detection of single-cell suspensions, significantly improves pigment yield and the genetic stability of the strains, optimizes the strain selection efficiency, and ensures the reliability of pigment production. Based on sustainability and economy, this invention performs crude pigment extraction on the optimal mutant strain to obtain a crude extract of mutant pigment. The maximum absorption wavelength is then determined using this extract. The optimal mutant strain is cultivated to obtain multiple optimal mutant strains. These mutant strains are then grouped according to a preset fermentation temperature to obtain multiple mutant experimental groups. The absorbance of each mutant experimental group is measured to obtain an absorbance value set. This absorbance value set is used to determine the optimal fermentation temperature. The absorbance value set includes the absorbance value of each mutant experimental group at the maximum absorption wavelength. Therefore, this invention significantly improves pigment yield and quality by crudely extracting pigment from the optimal mutant strain and determining the maximum absorption wavelength, further optimizing fermentation conditions to confirm the optimal fermentation temperature, thus providing key technical support for large-scale production and application. Next, this invention utilizes the optimal fermentation temperature and the optimal mutant strain for large-scale fermentation to obtain a high-secretion fermentation broth. The high-secretion fermentation broth is then subjected to cell wall disruption to obtain a cell wall-broken bacterial suspension. A dual-enzyme synergistic pigment release operation is performed on the cell wall-broken bacterial suspension to obtain an intracellular pigment solution. This intracellular pigment solution is then microencapsulated to obtain pigment microspheres. It is evident that this embodiment of the invention significantly improves pigment secretion efficiency and obtains a high-yield pigment fermentation broth by using the optimal mutant strain at the optimal fermentation temperature for large-scale fermentation. Furthermore, the cell wall disruption treatment and dual-enzyme synergistic release operation, combined with microencapsulation technology, efficiently extract intracellular pigments, improving pigment stability and bioavailability.Furthermore, this invention extracts a verification pigment extract from the pigment microspheres. After confirming that the verification pigment extract is the preset target pigment extract, the natural pigment is obtained using the verification pigment extract. It is evident that this embodiment of the invention ensures the accuracy and reliability of the extraction process by extracting the verification pigment extract from the pigment microspheres and confirming that it is the target pigment extract. This process not only improves the extraction efficiency of natural pigments but also ensures the purity and performance of the final natural pigment through strict quality control, making it fully compliant with the high standards required by the coatings industry for natural pigments. Therefore, this invention can solve the problems of low extraction efficiency, high energy consumption, and environmental pollution in the extraction of natural pigments for coatings. Attached Figure Description

[0087] Figure 1 This is a schematic flowchart of a method for extracting natural pigments for coatings based on microbial fermentation, provided in an embodiment of the present invention.

[0088] Figure 2 A functional block diagram of a natural pigment extraction system for coatings based on microbial fermentation provided in an embodiment of the present invention;

[0089] Figure 3 This is a schematic diagram of an electronic device for implementing the microbial fermentation-based method for extracting natural pigments for coatings, according to an embodiment of the present invention.

[0090] Explanation of reference numerals in the attached figures:

[0091] 10. Electronic device; 11. Processor; 12. Memory; 13. Bus.

[0092] The realization of the objective, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0093] It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0094] This application provides a method for extracting natural pigments for coatings based on microbial fermentation. The executing entity of this method includes, but is not limited to, at least one electronic device configured to execute the method provided in this application, such as a server or a terminal. In other words, the method can be executed by software or hardware installed on a terminal device or a server device, and the software may be a blockchain platform. The server includes, but is not limited to, a single server, a server cluster, a cloud server, or a cloud server cluster.

[0095] Reference Figure 1The diagram shown is a schematic flowchart of a method for extracting natural pigments for coatings based on microbial fermentation, according to an embodiment of the present invention. In this embodiment, the method for extracting natural pigments for coatings based on microbial fermentation includes:

[0096] S1. Obtain multiple initial strains, and use multiple initial strains and pre-constructed gene editing technology to obtain a gene recombinant strain group, wherein the gene recombinant strain group includes multiple gene recombinant strains.

[0097] It should be explained that obtaining multiple initial strains includes:

[0098] Multiple candidate strains were obtained using a pre-constructed marine microbial database;

[0099] Gene sequence alignment analysis was performed on all the candidate strains, resulting in multiple classification and attribution reports;

[0100] Based on the multiple classification and attribution reports, multiple target strains were identified from multiple candidate strains;

[0101] Each of the multiple target strains was cultured in pre-confirmed dried seawater medium to obtain multiple stable target strains, with each target strain corresponding to a stable target strain.

[0102] Intracellular pigment content was detected for each of the multiple stable target strains to obtain a set of intracellular pigment content values. The set of intracellular pigment content values ​​included multiple intracellular pigment content values, and each intracellular pigment content value corresponded one-to-one with a stable target strain.

[0103] The stable target strains corresponding to intracellular pigment content values ​​that are higher than the preset intracellular pigment content threshold are identified as initial strains, resulting in multiple initial strains.

[0104] Furthermore, the pre-constructed marine microbial database is a microbial library integrating marine microbial strains, containing a large amount of data on the genetic information, growth characteristics, and metabolites of marine microorganisms, used to screen strains with specific characteristics. Optionally, a marine microbial gene database can be used as the marine microbial database. Candidate strains are strains that are initially screened from the marine microbial database and may have the potential for target pigment synthesis. Gene sequence alignment analysis involves BLASTing the 16S rRNA gene sequences of candidate strains to obtain information on similar pattern strains. Optionally, a biomolecular sequence alignment search tool can be used as the method for gene sequence alignment analysis. The taxonomic classification report includes the taxonomic position and similarity score of the candidate strains. Taxonomic position refers to the position of the strain in the biological classification system, and the similarity score reflects the degree of similarity between its gene sequence and known strains.

[0105] Understandably, identifying multiple target strains from multiple candidate strains based on the aforementioned multiple classification reports refers to selecting candidate strains with a similarity ≥97% to known pigment-producing strains, while excluding pathogenic bacteria or strains that are difficult to culture. The aim is to screen strains with a high probability of synthesizing the target pigment and that are easy to culture. The pre-confirmed aged seawater culture medium is a medium composed of artificial seawater supplemented with 0.5% peptone and 0.1% yeast extract, with a pH of 7.5±0.2, used to simulate the marine environment and ensure the growth adaptability of the target strains. The purpose of using aged seawater culture medium is to provide suitable growth conditions for the strains, enabling them to grow stably and exhibit their pigment synthesis ability. Stable target strains are strains that maintain stable colony morphology, growth rate (generation time difference ≤10%), and pigment production (fluctuation ≤15%) after three consecutive passages in aged seawater culture medium, i.e., the cultured target strains do not exhibit gene mutations. Intracellular pigment content detection is a method used to determine the content of pigments within the strain cells; optionally, high-performance liquid chromatography (HPLC) is used as the intracellular pigment content detection method. Intracellular pigment content is the specific amount of pigment within a stable target strain, obtained through intracellular pigment content detection. The intracellular pigment content set is a collection of multiple intracellular pigment content values. The preset intracellular pigment content threshold is a standard value pre-set according to experimental requirements to screen for strains with higher intracellular pigment contents. The initial strain is a stable target strain with an intracellular pigment content value higher than the intracellular pigment content threshold.

[0106] It should be explained that the method of obtaining a gene recombinant strain population using multiple initial strains and pre-constructed gene editing technology includes:

[0107] For each of the multiple initial strains, the following procedure was performed:

[0108] The initial strain was physically isolated to obtain experimental strains and control strains;

[0109] Whole-genome sequencing was performed on the experimental strain to obtain its genomic information;

[0110] The experimental strain was recombined using its genomic information and pre-constructed gene editing technology to obtain a primary gene-recombined strain.

[0111] The primary gene recombinant strain and the control strain were fermented separately to obtain recombinant fermentation cell liquid and control fermentation cell liquid, respectively.

[0112] The recombinant fermentation cell broth and the control fermentation cell broth were subjected to crude pigment extraction to obtain crude recombinant pigment extract and crude control pigment extract, respectively.

[0113] Obtain the pigment mass concentration of the crude extract of recombinant pigment and the pigment mass concentration of the crude extract of control pigment;

[0114] The yield increase rate was calculated using the pigment concentration of the recombinant pigment crude extract and the pigment concentration of the control pigment crude extract.

[0115] The yield increase rate is compared with the preset yield increase threshold, and the primary gene recombinant strains corresponding to the yield increase rate that is greater than or equal to the yield increase threshold are identified as gene recombinant strains.

[0116] Multiple recombinant strains were combined to obtain a group of recombinant strains.

[0117] Further, physical isolation involves streaking the initial strain onto a solid culture medium and separating single colonies into experimental or control strains. The experimental strain is the strain that will subsequently undergo gene recombination. The control strain is homologous to the experimental strain and is used to compare yield increases. Whole-genome sequencing involves sequencing the entire genome of the experimental strain to obtain its complete gene sequence information. The aim is to gain a detailed understanding of the strain's gene composition and structure, providing fundamental data for subsequent gene editing.

[0118] It should be explained that the experimental strain's genomic information is the gene sequence information obtained through whole-genome sequencing, including gene location, sequence, and function. The pre-constructed gene editing technology is a technique used to edit the genes of the experimental strain; optionally, gene editing is used as the gene editing technology. Gene recombination utilizes gene editing technology to modify the genome of the experimental strain, giving it new gene combinations or altering the function of existing genes, potentially improving pigment synthesis capacity. The primary gene recombination strain is the strain obtained after gene recombination. Fermentation treatment involves culturing the strain under specific fermentation conditions, allowing it to grow, reproduce, and synthesize metabolites such as pigments. Fermentation conditions include temperature, pH, and nutrients. The recombinant fermentation broth is the fermentation liquid obtained after fermentation treatment of the primary gene recombination strain. The control fermentation broth is the fermentation broth obtained after fermentation treatment of the control strain, used for comparative analysis with the recombinant fermentation broth.

[0119] Further, crude pigment extraction is the process of extracting pigment from the fermentation broth; optionally, ultrasound-assisted extraction is used as the method for crude pigment extraction. The recombinant pigment crude extract is a pigment-containing liquid extracted from the recombinant fermentation cell broth. The control pigment crude extract is a pigment-containing liquid extracted from the control fermentation cell broth. The pigment mass concentration of the recombinant pigment crude extract is the mass concentration of pigment in the recombinant pigment crude extract. The pigment mass concentration of the control pigment crude extract is the mass concentration of pigment in the control pigment crude extract. The pigment mass concentrations of the recombinant pigment crude extract and the control pigment crude extract can be determined using spectrophotometry. The yield increase rate is a value calculated using the pigment mass concentrations of the recombinant pigment crude extract and the control pigment crude extract to evaluate the impact of gene recombination on pigment yield.

[0120] The calculation formula is as follows:

[0121]

[0122] Where P represents the rate of increase in output, and C E C represents the pigment mass concentration of the crude extract of recombinant pigments. B This indicates the pigment concentration of the control pigment crude extract.

[0123] Understandably, the preset yield increase threshold is a standard value pre-set based on the experimental objectives to determine whether the recombinant strain has achieved the expected yield increase effect. A recombinant strain is a primary recombinant strain whose yield increase rate is greater than or equal to the yield increase threshold. A recombinant strain population is a group of strains obtained by aggregating multiple recombinant strains.

[0124] S2. For each of the recombinant strains in the recombinant strain group, a single-cell suspension is prepared to obtain a single-cell suspension set. The single-cell suspension set includes multiple single-cell suspensions, and each single-cell suspension corresponds one-to-one with a recombinant strain. Multiple single-cell suspensions are subjected to fluorescence detection to obtain multiple fluorescence detection nodes. The optimal mutant strain is identified using multiple fluorescence detection nodes.

[0125] It should be explained that the fluorescence detection of multiple single-cell suspensions to obtain multiple fluorescence detection nodes includes:

[0126] Perform the following operation on each of the multiple single-cell suspensions:

[0127] The single-cell suspension was introduced into a pre-constructed nanoreactor to obtain an initial nanoreactor;

[0128] A pre-confirmed fluorescent reporter gene expression vector was added to the initial nanoreactor to obtain the labeled nanoreactor;

[0129] The labeled nanoreactors were placed in a pre-confirmed constant temperature environment and cultured for 48 hours to obtain labeled microculture medium. Every 4 hours within 48 hours, the cell fluorescence density of the labeled nanoreactors was detected using a pre-confirmed high-throughput fluorescence microscope to obtain cell density sequences and fluorescence intensity sequences.

[0130] Cell density-time curves were constructed using cell density sequences, and the maximum specific growth rate was obtained based on the cell density-time curves.

[0131] The maximum pigment synthesis activity index was obtained based on cell density and fluorescence intensity sequences.

[0132] The coefficient of variation was calculated from the fluorescence intensity sequence to obtain the coefficient of variation.

[0133] The maximum specific growth rate, maximum pigment synthesis activity index, and coefficient of variation are summarized to obtain the fluorescence detection node;

[0134] By summarizing the fluorescence detection nodes, multiple fluorescence detection nodes are obtained, and each fluorescence detection node corresponds one-to-one with a labeled microculture medium.

[0135] Furthermore, the single-cell suspension is formed by dispersing recombinant bacterial strains in a liquid culture medium, resulting in a suspension of single cells in the liquid. This is to facilitate subsequent fluorescence detection and analysis. The pre-constructed nanoreactor is a reaction device used for cell culture and detection in a small volume. Nanoreactors are existing technology and will not be described in detail here. The initial nanoreactor is the nanoreactor obtained after introducing the single-cell suspension.

[0136] Understandably, a pre-validated fluorescent reporter gene expression vector is a plasmid containing a fluorescent reporter gene for labeling cells and enabling them to express fluorescent proteins under specific conditions. Optionally, green fluorescent protein is used as the fluorescent reporter gene expression vector. The labeled nanoreactor is a nanoreactor in which cells are labeled after the fluorescent reporter gene expression vector is added to the initial nanoreactor. The pre-validated isothermal environment is a pre-set and validated culture condition to ensure cell growth and expression of fluorescent proteins at a stable temperature. Optionally, an incubator is used as the isothermal environment. The labeled micromedium is a micromedium in which cells grow and express fluorescent proteins after 48 hours of isothermal incubation. The pre-validated high-throughput fluorescence microscope is a microscope device capable of rapidly and sensitively detecting fluorescence signals for observing and recording cell fluorescence density. Optionally, a fully automated fluorescence microscope is used as the high-throughput fluorescence microscope. Cell fluorescence density detection is a method for quantitatively detecting cell density and fluorescence intensity using a high-throughput fluorescence microscope. Optionally, a fully automated fluorescence microscope is used as the instrument for cell fluorescence density detection.

[0137] It should be explained that a cell density sequence refers to a sequence obtained by measuring cell density every 4 hours over a 48-hour period, obtaining multiple cell density values, and then arranging these values ​​in chronological order. A fluorescence intensity sequence is obtained by measuring fluorescence intensity every 4 hours over a 48-hour period, obtaining multiple fluorescence intensity values, and then arranging these values ​​in chronological order. A cell density-time curve is a curve plotted based on the cell density sequence, with time on the horizontal axis and cell density on the vertical axis, used to analyze the specific growth rate of cells. Obtaining the maximum specific growth rate based on the cell density-time curve refers to calculating the maximum specific growth rate reached by cells during growth. Specific growth rate refers to the rate of increase in the number of cells per unit time. The maximum pigment synthesis activity index is the maximum fluorescence intensity per unit cell density, reflecting the maximum activity of cells in synthesizing pigments during growth. The coefficient of variation is the ratio of the standard deviation to the mean of the fluorescence intensity sequence, used to assess the degree of variation in fluorescence intensity and reflect the homogeneity of the cell population.

[0138] It should be explained that the identification of the optimal mutant strain using multiple fluorescence detection nodes includes:

[0139] The maximum specific growth rate, maximum pigment synthesis activity index, and coefficient of variation in multiple fluorescence detection nodes were normalized to obtain multiple normalized fluorescence detection nodes. The normalized fluorescence detection nodes include the normalized maximum specific growth rate, the normalized maximum pigment synthesis activity index, and the normalized coefficient of variation.

[0140] A comprehensive evaluation value is calculated using each of the multiple normalized fluorescence detection nodes and a pre-constructed comprehensive evaluation value calculation formula to obtain a comprehensive evaluation value set. The comprehensive evaluation value calculation formula is as follows:

[0141] S = 0.4 × T max +0.5×L+0.1×(1-CV)

[0142] Where S represents the comprehensive evaluation value, T max denoted as the normalized maximum specific growth rate, L represents the normalized maximum pigment synthesis activity index, and CV represents the normalized coefficient of variation.

[0143] The comprehensive evaluation values ​​in the comprehensive evaluation value set are arranged in descending order to obtain a comprehensive evaluation value sequence. The top 5 comprehensive evaluation values ​​are extracted from the comprehensive evaluation value sequence. The labeled micro-culture media corresponding to the extracted 5 comprehensive evaluation values ​​are all identified as candidate micro-culture media, thus obtaining 5 candidate culture media.

[0144] Verification experiments were conducted on each of the five candidate culture media, resulting in five experimental reports. Based on these reports, the optimal labeled microculture medium was determined, and the gene recombinant strain corresponding to the optimal labeled microculture medium was identified as the optimal mutant strain.

[0145] Furthermore, the maximum specific growth rate, maximum pigment synthesis activity index, and coefficient of variation for multiple fluorescence detection nodes are normalized. This involves extracting the maximum specific growth rate, maximum pigment synthesis activity index, and coefficient of variation from multiple fluorescence detection nodes to obtain sets of maximum specific growth rates, maximum pigment synthesis activity indices, and coefficients of variation. The set of maximum specific growth rates includes multiple maximum specific growth rates, the set of maximum pigment synthesis activity indices includes multiple maximum pigment synthesis activity indices, and the set of coefficients of variation includes multiple coefficients of variation. Normalization is then performed on these multiple maximum specific growth rates, maximum pigment synthesis activity indices, and coefficients of variation to obtain multiple normalized maximum specific growth rates, multiple normalized maximum pigment synthesis activity indices, and multiple normalized coefficients of variation. Each normalized maximum specific growth rate, each normalized maximum pigment synthesis activity index, and each normalized coefficient of variation is then mapped back to its corresponding fluorescence detection node to obtain multiple normalized fluorescence detection nodes. Optionally, Min-Max normalization can be used as the normalization method.

[0146] Understandably, a normalized fluorescence detection node includes the normalized maximum specific growth rate, the normalized maximum pigment synthesis activity index, and the normalized coefficient of variation. The normalized maximum specific growth rate is the normalized value of the maximum specific growth rate. The normalized maximum pigment synthesis activity index is the normalized value of the maximum pigment synthesis activity index. The normalized coefficient of variation is the normalized value of the coefficient of variation. The comprehensive evaluation value is calculated using a pre-constructed comprehensive evaluation value calculation formula. The comprehensive evaluation value set is a collection of comprehensive evaluation values ​​from multiple fluorescence detection nodes. A confirmatory experiment is a validation experiment conducted on candidate micromediums to verify whether their performance meets expectations. For example, five candidate micromediums are recultured to obtain five recultured candidate micromediums, and cell fluorescence density is detected again for each of the five recultured micromediums. The experimental report is a record of the validation experiment containing experimental data and analytical conclusions. Identifying the optimal labeled micromedium based on the five experimental reports means determining the labeled micromedium with the highest comprehensive evaluation value from the five candidate micromediums based on the results of the confirmatory experiment. The optimal mutant strain is the recombinant strain corresponding to the optimal labeled microculture medium.

[0147] S3. Perform crude pigment extraction on the best mutant strain to obtain a crude extract of mutant pigment, and use the crude extract of mutant pigment to determine the maximum absorption wavelength.

[0148] It should be explained that the process of crudely extracting pigments from the optimal mutant strain to obtain a crude extract of mutant pigments, and using this crude extract to determine the maximum absorption wavelength, includes:

[0149] The optimal mutant strain was fermented to obtain the mutant strain fermentation broth, and the mutant strain fermentation broth was centrifuged to obtain cell precipitate;

[0150] The bacterial cell precipitate was resuspended to obtain a bacterial cell suspension, and the bacterial cell suspension was then broken down to obtain a cell lysate.

[0151] The cell lysate was subjected to static extraction to obtain an extraction mixture. The extraction mixture was then centrifuged to obtain a supernatant, which was used as the crude extract of pigments from the mutant strain.

[0152] Absorption spectral data were obtained by scanning the crude pigment extract of the mutant strain using a pre-confirmed UV-Vis spectrophotometer across the entire wavelength range.

[0153] Peak identification is performed on the absorption spectrum data to obtain the maximum absorption wavelength.

[0154] Further, the mutant strain fermentation broth is the fermentation broth of the optimal mutant strain obtained after fermentation treatment. Centrifugation is the operation of using centrifugal force to precipitate the bacterial cells in the fermentation broth to the bottom of the centrifuge tube. The bacterial cell precipitate is the portion of bacterial cells deposited at the bottom of the centrifuge tube after centrifugation. Resuspension is the process of resuspending the centrifuged bacterial cell precipitate in a liquid such as buffer or physiological saline for subsequent disruption treatment. The bacterial cell suspension is the liquid obtained after resuspension treatment, in which the bacterial cells are uniformly suspended in the liquid. Disruption treatment is the process of physically breaking down the bacterial cell walls and cell membranes to release intracellular substances (such as pigments). Optionally, ultrasonic disruption is used as the disruption treatment method. The cell disruption solution is the liquid obtained after disruption treatment. Static extraction is the process of allowing the cell disruption solution to stand for a period of time to separate the target substances such as pigments from cell debris and other impurities. The extraction mixture is the liquid containing the pigments dissolved therein obtained after static extraction. Centrifugation is the process of using centrifugal force to separate the pigments from impurities in the extraction mixture. The supernatant is the liquid mainly containing dissolved pigments that remains on the top layer after centrifugation. The mutant strain pigment extract is a liquid containing pigment obtained after the above treatment.

[0155] Understandably, a pre-confirmed UV-Vis spectrophotometer is an instrument used to measure the absorption spectra of crude extracts of mutant pigments in the UV and visible regions. UV-Vis spectrophotometers are existing technology and will not be described in detail here. Full-wavelength scanning is the operation of continuously measuring the absorbance of the crude extract of mutant pigments from the UV to the visible region using a UV-Vis spectrophotometer to obtain the absorption spectrum data of the crude extract. The absorption spectrum data is a series of absorbance values ​​obtained after a full-wavelength scan. Peak identification is the identification of the point with the highest absorbance from the absorption spectrum data. Optionally, peak detection is used as the method for peak identification. The maximum absorption wavelength is the wavelength corresponding to the point with the highest absorbance in the absorption spectrum.

[0156] S4. Cultivate the best mutant strain to obtain multiple best mutant strains. Group the multiple best mutant strains according to the preset fermentation temperature to obtain multiple mutant strain experimental groups. Measure the absorbance of each mutant strain experimental group in the multiple mutant strain experimental groups to obtain an absorbance value set. Use the absorbance value set to determine the optimal fermentation temperature. The absorbance value set includes the absorbance value of each mutant strain experimental group at the maximum absorption wavelength.

[0157] It should be explained that the process of measuring absorbance for each of the multiple mutant experimental groups to obtain a set of absorbance values, and then using this set of absorbance values ​​to determine the optimal fermentation temperature, includes:

[0158] Each mutant experimental group in the multiple mutant experimental groups was inoculated into a pre-confirmed fermentation medium and cultured to obtain a fermentation broth sample set, wherein the fermentation broth sample set includes multiple fermentation broth samples, and the fermentation broth samples correspond one-to-one with the mutant experimental groups.

[0159] Multiple fermentation broth samples were pretreated to obtain a supernatant sample set, which included multiple supernatant samples.

[0160] The absorbance of multiple supernatant samples was measured using the maximum absorption wavelength and a UV-Vis spectrophotometer to obtain a set of absorbance values.

[0161] Construct a temperature-absorbance curve using the absorbance value set;

[0162] The optimal fermentation temperature was determined using the temperature-absorbance curve.

[0163] Further, the optimal mutant strains are cultivated to obtain multiple optimal mutant strains. This refers to cultivating the optimal mutant strains using the streak plate method to obtain multiple optimal mutant strains. The preset fermentation temperature is a series of different fermentation temperatures pre-set according to the experimental design to evaluate the effect of different temperatures on pigment yield. For example, 25℃, 30℃, 35℃, and 40℃. The mutant experimental group is an experimental group obtained by grouping multiple optimal mutant strains according to the preset fermentation temperature. The pre-validated fermentation medium is a pre-validated culture medium. The fermentation broth sample set is a collection containing multiple fermentation broth samples. The fermentation broth sample is the liquid extracted from each cultured mutant experimental group for subsequent analysis. Pretreatment is an operation to remove impurities from the fermentation broth sample to facilitate subsequent absorbance measurement. The supernatant sample set is a collection containing multiple supernatant samples. The supernatant sample is the supernatant obtained after pretreatment. Absorbance measurement is the process of measuring the absorbance of the supernatant sample at the maximum absorption wavelength using a UV-Vis spectrophotometer. The absorbance value set is a collection of absorbance values ​​from multiple supernatant samples. The temperature-absorbance curve is plotted with fermentation temperature on the horizontal axis and absorbance value on the vertical axis. The optimal fermentation temperature is determined using the temperature-absorbance curve; it represents the fermentation temperature at which the absorbance value reaches its maximum.

[0164] S5. Scale up fermentation using the optimal fermentation temperature and the optimal mutant strain to obtain a high-secretion fermentation broth.

[0165] It should be explained that large-scale fermentation is the process of conducting fermentation on a large scale after determining the optimal fermentation temperature and the optimal mutant strain. High-secretion fermentation broth is the fermentation broth obtained after large-scale fermentation.

[0166] S6. The high-secretion fermentation broth is subjected to cell wall disruption treatment to obtain a cell wall disruption suspension. A dual-enzyme synergistic pigment release operation is performed on the cell wall disruption suspension to obtain an intracellular pigment solution. The intracellular pigment solution is microencapsulated to obtain pigment microspheres.

[0167] It should be explained that the process of microencapsulating the intracellular pigment fluid to obtain pigment microspheres includes:

[0168] The intracellular pigment solution and the pre-confirmed aerogel-sodium alginate composite emulsion were homogenized and emulsified at a preset homogenization speed to obtain a primary emulsion.

[0169] The primary emulsion was ion-gelled to obtain multiple primary microspheres;

[0170] Each primary microsphere in a plurality of primary microspheres was cross-linked and embedded in a pre-confirmed chitosan buffer to obtain a plurality of double-embedded wet microspheres;

[0171] Multiple double-embedded wet microspheres were freeze-dried under vacuum to obtain multiple original pigment microspheres;

[0172] Random sampling was performed on multiple original pigment microspheres to obtain a sampled original pigment microsphere set and a set of undetermined original pigment microspheres. The sampled original pigment microsphere set included multiple sampled original pigment microspheres, and the set of undetermined original pigment microspheres included multiple undetermined original pigment microspheres.

[0173] The pigment embedding rate of multiple sampled original pigment microspheres was detected using a pre-confirmed pigment embedding rate detector, resulting in a set of pigment embedding rate detection values, which included multiple pigment embedding rate detection values.

[0174] Multiple pigment embedding rate detection values ​​are compared with preset pigment embedding rate detection thresholds. Sampled original pigment microspheres with pigment embedding rate detection values ​​greater than or equal to the pigment embedding rate detection thresholds are identified as qualified original pigment microspheres. The qualified original pigment microspheres and sampled original pigment microspheres are used to obtain the original pigment microsphere qualification rate.

[0175] The pass rate of the original pigment microspheres is compared with the preset pass rate threshold of the original pigment microspheres. If the pass rate of the original pigment microspheres is less than the threshold, the homogenization speed is adjusted and the adjusted homogenization speed is used as the preset homogenization speed. The process is repeated until the pass rate of the original pigment microspheres is greater than or equal to the threshold. The multiple undetermined original pigment microspheres are then confirmed as pigment microspheres.

[0176] Further, the cell wall-broken bacterial suspension is a liquid in which intracellular substances are released into the suspension after cell wall disruption. Dual-enzyme synergistic pigment release is an operation performed on the cell wall-broken bacterial suspension to more efficiently release pigments from it. Optionally, a composite enzyme synergistic effect can be used as the method for dual-enzyme synergistic pigment release. Intracellular pigment solution is a solution mainly composed of pigments released from the bacterial cells after the dual-enzyme synergistic pigment release operation. The pre-confirmed aerogel-sodium alginate composite emulsion is a pre-confirmed emulsion composed of aerogel and sodium alginate used in the microencapsulation process. The preset homogenization speed is a stirring speed pre-set during homogenization emulsification, and the level of the homogenization speed affects the pigment encapsulation rate of the pigment microspheres. Therefore, in subsequent processing steps, the homogenization speed of this embodiment will be adjusted to obtain pigment microspheres with a qualified pigment encapsulation rate. Homogenization is the process of mixing intracellular pigment solution with aerogel-sodium alginate composite emulsion at a preset homogenization speed to obtain a uniform emulsion.

[0177] Understandably, ionogel solidification is a process in which sodium alginate in a primary emulsion forms a gel through ion cross-linking, and is further solidified into microspheres. Primary microspheres are the initial pigment microspheres formed after ionogel solidification. The pre-validated chitosan buffer is a pre-verified and validated buffer for further encapsulating the microspheres. Cross-linking encapsulation involves placing the primary microspheres in the chitosan buffer to allow chitosan to cross-link with the microsphere surface, forming a bilayer encapsulation structure. This aims to improve the stability of the microspheres and the pigment encapsulation rate. After cross-linking encapsulation, the surface of the double-encapsulated wet microspheres is encapsulated by a chitosan layer, forming a bilayer structure. Vacuum freeze-drying is the process of freezing the double-encapsulated wet microspheres at low temperature and then drying them under vacuum conditions. The purpose is to avoid damage to the pigment from high temperatures while maintaining the structural integrity of the microspheres. The original pigment microspheres are the dried microspheres obtained after vacuum freeze-drying.

[0178] Further, the sampled original pigment microsphere set is a collection containing multiple sampled original pigment microspheres. These sampled original pigment microspheres are randomly selected from multiple original pigment microspheres for pigment embedding rate detection. The undetermined original pigment microsphere set is a collection containing multiple undetermined original pigment microspheres. These undetermined original pigment microspheres are those for which pigment embedding rate detection is not required. The pre-confirmed pigment embedding rate detector is an instrument used to detect the pigment embedding rate in the microspheres. Optionally, a UV-Vis spectrophotometer is used as the pigment embedding rate detector. Pigment embedding rate detection is the process of using a pigment embedding rate detector to detect the sampled original pigment microspheres, with the aim of confirming the pigment embedding rate of the sampled original pigment microspheres. The pigment embedding rate detection value set is a collection containing multiple pigment embedding rate values. The pigment embedding rate detection value is the pigment embedding rate detection value of the sampled original pigment microspheres. The preset pigment embedding rate detection threshold is a pre-set minimum standard value for pigment embedding rate. When the pigment encapsulation rate is greater than or equal to the pigment encapsulation rate detection threshold, the sampled original pigment microspheres are considered qualified. For example, 95%. Qualified original pigment microspheres are those sampled original pigment microspheres whose pigment encapsulation rate is greater than or equal to the preset threshold.

[0179] It should be explained that the original pigment microsphere pass rate is the proportion of qualified original pigment microspheres to the total number of sampled original pigment microspheres. The preset original pigment microsphere pass rate threshold is a pre-set minimum standard value for the original pigment microsphere pass rate. If the original pigment microsphere pass rate is lower than this threshold, the homogenization speed needs to be adjusted. Adjusting the homogenization speed refers to adjusting the stirring speed during the homogenization emulsification process based on the original pigment microsphere pass rate results to optimize the pigment microsphere preparation conditions. For example, if the preset homogenization speed is 1200 rpm, and the original pigment microsphere pass rate is lower than the threshold, the homogenization speed is increased, for example, to 1250 rpm. The pigment microspheres are the undetermined original pigment microspheres corresponding to the original pigment microsphere pass rate being greater than or equal to the threshold after the above microencapsulation process.

[0180] S7. Extract the verification pigment extract from the pigment microspheres. After confirming that the verification pigment extract is the preset target pigment extract, use the verification pigment extract to obtain the natural pigment.

[0181] It should be explained that, after confirming that the verification pigment extract is the preset target pigment extract, obtaining the natural pigment using the verification pigment extract includes:

[0182] The pigment extract was mixed with the pre-confirmed acrylic resin emulsion to obtain the paint to be tested.

[0183] The pre-confirmed aluminum plate was sprayed with the color paint to be tested to obtain the sprayed aluminum plate.

[0184] Color difference detection is performed on the coated aluminum plate to obtain the color difference detection value. The color difference detection value is compared with the preset color difference detection threshold. If the color difference detection value is less than the color difference detection threshold, the verification pigment extract is confirmed as the target pigment extract.

[0185] The target pigment extract was purified to obtain the natural pigment.

[0186] Further, the verification pigment extract is a liquid containing the target pigment extracted from pigment microspheres. The pre-confirmed acrylic resin emulsion is an emulsion whose main component is acrylic resin, used in the preparation of coatings and paints. The test paint is a paint obtained by mixing the verification pigment extract with the pre-confirmed acrylic resin emulsion, used for subsequent spraying and color difference detection. The sprayed aluminum plate is a sample obtained after the test paint is uniformly sprayed onto the pre-confirmed aluminum plate. Color difference detection is a method of measuring the color of the sprayed aluminum plate with a colorimeter to evaluate the difference between the color of the paint and the standard color. Optionally, a colorimeter is used as the instrument for color difference detection. The color difference detection value is the absolute difference between the color of the sprayed aluminum plate and the standard color, measured by the colorimeter. The preset color difference detection threshold is a pre-set standard value used to judge the magnitude of the color difference. For example, a color difference detection value less than 1.0 is considered acceptable. The preset target pigment extract is a pigment extract that is pre-set according to the experimental design and is less than the color difference detection threshold in the color difference detection. Purification is a process used to remove impurities from a verified pigment extract, identifying it as a target pigment extract, thereby increasing its purity. Natural pigments are high-purity pigments obtained through purification and can be used directly in coatings.

[0187] To address the problems described in the background art, this invention obtains multiple initial bacterial strains and utilizes pre-constructed gene editing technology to obtain a recombinant bacterial strain population. The recombinant bacterial strain population includes multiple recombinant bacterial strains. For each recombinant bacterial strain in the population, single-cell suspensions are prepared to obtain a set of single-cell suspensions. Each single-cell suspension set includes multiple single-cell suspensions, and each single-cell suspension corresponds one-to-one with a recombinant bacterial strain. Fluorescence detection is performed on multiple single-cell suspensions to obtain multiple fluorescence detection nodes. The optimal mutant strain is identified using these multiple fluorescence detection nodes. Therefore, this invention, by obtaining multiple initial bacterial strains, constructing a recombinant bacterial strain population using gene editing technology, and then screening for the optimal mutant strain through fluorescence detection of single-cell suspensions, significantly improves pigment yield and the genetic stability of the strains, optimizes the strain selection efficiency, and ensures the reliability of pigment production. Based on sustainability and economy, this invention performs crude pigment extraction on the optimal mutant strain to obtain a crude extract of mutant pigment. The maximum absorption wavelength is then determined using this extract. The optimal mutant strain is cultivated to obtain multiple optimal mutant strains. These mutant strains are then grouped according to a preset fermentation temperature to obtain multiple mutant experimental groups. The absorbance of each mutant experimental group is measured to obtain an absorbance value set. This absorbance value set is used to determine the optimal fermentation temperature. The absorbance value set includes the absorbance value of each mutant experimental group at the maximum absorption wavelength. Therefore, this invention significantly improves pigment yield and quality by crudely extracting pigment from the optimal mutant strain and determining the maximum absorption wavelength, further optimizing fermentation conditions to confirm the optimal fermentation temperature, thus providing key technical support for large-scale production and application. Next, this invention utilizes the optimal fermentation temperature and the optimal mutant strain for large-scale fermentation to obtain a high-secretion fermentation broth. The high-secretion fermentation broth is then subjected to cell wall disruption to obtain a cell wall-broken bacterial suspension. A dual-enzyme synergistic pigment release operation is performed on the cell wall-broken bacterial suspension to obtain an intracellular pigment solution. This intracellular pigment solution is then microencapsulated to obtain pigment microspheres. It is evident that this embodiment of the invention significantly improves pigment secretion efficiency and obtains a high-yield pigment fermentation broth by using the optimal mutant strain at the optimal fermentation temperature for large-scale fermentation. Furthermore, the cell wall disruption treatment and dual-enzyme synergistic release operation, combined with microencapsulation technology, efficiently extract intracellular pigments, improving pigment stability and bioavailability.Furthermore, this invention extracts a verification pigment extract from the pigment microspheres. After confirming that the verification pigment extract is the preset target pigment extract, the natural pigment is obtained using the verification pigment extract. It is evident that this embodiment of the invention ensures the accuracy and reliability of the extraction process by extracting the verification pigment extract from the pigment microspheres and confirming that it is the target pigment extract. This process not only improves the extraction efficiency of natural pigments but also ensures the purity and performance of the final natural pigment through strict quality control, making it fully compliant with the high standards required by the coatings industry for natural pigments. Therefore, this invention can solve the problems of low extraction efficiency, high energy consumption, and environmental pollution in the extraction of natural pigments for coatings.

[0188] like Figure 2 The diagram shown is a functional block diagram of a natural pigment extraction system for coatings based on microbial fermentation provided in an embodiment of the present invention.

[0189] The microbial fermentation-based natural pigment extraction system 100 for coatings described in this invention can be installed in an electronic device. Depending on the functions implemented, the microbial fermentation-based natural pigment extraction system 100 may include a mutant strain screening module 101, a fermentation condition confirmation module 102, a pigment microsphere synthesis module 103, and a natural pigment verification module 104. The module described in this invention can also be referred to as a unit, which refers to a series of computer program segments that can be executed by the processor of an electronic device and can perform a fixed function, and which are stored in the memory of the electronic device.

[0190] The mutant strain screening module 101 is used to obtain multiple initial strains and to obtain a gene recombinant strain group using multiple initial strains and pre-constructed gene editing technology, wherein the gene recombinant strain group includes multiple gene recombinant strains.

[0191] For each of the recombinant strains in the aforementioned recombinant strain group, a single-cell suspension was prepared to obtain a single-cell suspension set. The single-cell suspension set included multiple single-cell suspensions, and each single-cell suspension corresponded one-to-one with a recombinant strain. Multiple single-cell suspensions were subjected to fluorescence detection to obtain multiple fluorescence detection nodes. The optimal mutant strain was identified using multiple fluorescence detection nodes.

[0192] The fermentation condition confirmation module 102 is used to perform crude pigment extraction on the optimal mutant strain to obtain a crude pigment extract of the mutant strain, and to confirm the maximum absorption wavelength using the crude pigment extract of the mutant strain.

[0193] The best mutant strains were cultivated to obtain multiple best mutant strains. The multiple best mutant strains were grouped according to the preset fermentation temperature to obtain multiple mutant strain experimental groups. The absorbance of each mutant strain experimental group in the multiple mutant strain experimental groups was measured to obtain an absorbance value set. The optimal fermentation temperature was determined using the absorbance value set. The absorbance value set includes the absorbance value of each mutant strain experimental group in the multiple mutant strain experimental groups at the maximum absorption wavelength.

[0194] The pigment microsphere synthesis module 103 is used to carry out large-scale fermentation using the optimal fermentation temperature and the optimal mutant strain to obtain a high-secretion fermentation broth.

[0195] The high-secretion fermentation broth was subjected to cell wall disruption to obtain a cell wall disruption suspension. A dual-enzyme synergistic pigment release operation was performed on the cell wall disruption suspension to obtain an intracellular pigment solution. The intracellular pigment solution was then microencapsulated to obtain pigment microspheres.

[0196] The natural pigment verification module 104 is used to extract a verification pigment extract from the pigment microspheres, and after confirming that the verification pigment extract is the preset target pigment extract, the natural pigment is obtained using the verification pigment extract.

[0197] In detail, the modules in the microbial fermentation-based natural pigment extraction system 100 for coatings described in this embodiment of the invention employ the same methods as described above during use. Figure 1 The method used is the same as the natural pigment extraction method for coatings based on microbial fermentation described in the article, and can produce the same technical effect, so it will not be repeated here.

[0198] like Figure 3 The diagram shown is a schematic representation of an electronic device for implementing a method for extracting natural pigments for coatings based on microbial fermentation, according to an embodiment of the present invention.

[0199] The electronic device 1 may include a processor 10, a memory 11 and a bus 12, and may also include a computer program stored in the memory 11 and executable on the processor 10, such as a method program for extracting natural pigments for coatings based on microbial fermentation.

[0200] The memory 11 includes at least one type of readable storage medium, such as flash memory, portable hard drive, multimedia card, card-type memory (e.g., SD or DX memory), magnetic memory, magnetic disk, optical disk, etc. In some embodiments, the memory 11 can be an internal storage unit of the electronic device 1, such as a portable hard drive. In other embodiments, the memory 11 can be an external storage device of the electronic device 1, such as a plug-in portable hard drive, smart media card (SMC), secure digital card (SD), flash card, etc., equipped on the electronic device 1. Furthermore, the memory 11 includes both internal storage units and external storage devices of the electronic device 1. The memory 11 can be used not only to store application software and various types of data installed on the electronic device 1, such as the code of a method for extracting natural pigments for coatings based on microbial fermentation, but also to temporarily store data that has been output or will be output.

[0201] In some embodiments, the processor 10 may be composed of integrated circuits, such as a single packaged integrated circuit or multiple integrated circuits with the same or different functions, including combinations of one or more central processing units (CPUs), microprocessors, digital processing chips, graphics processors, and various control chips. The processor 10 is the control unit of the electronic device, connecting various components of the entire electronic device through various interfaces and lines. It executes programs or modules stored in the memory 11 (e.g., a method for extracting natural pigments for coatings based on microbial fermentation) and calls data stored in the memory 11 to perform various functions of the electronic device 1 and process data.

[0202] The bus 12 can be a peripheral component interconnect (PCI) bus or an extended industry standard architecture (EISA) bus, etc. The bus 12 can be divided into an address bus, a data bus, a control bus, etc. The bus 12 is configured to realize the connection and communication between the memory 11 and at least one processor 10, etc.

[0203] Figure 3 Only electronic devices with components are shown; it will be understood by those skilled in the art that... Figure 3The structure shown does not constitute a limitation on the electronic device 1, and may include fewer or more components than shown, or combine certain components, or have different component arrangements.

[0204] For example, although not shown, the electronic device 1 may also include a power supply (such as a battery) to power various components. Preferably, the power supply can be logically connected to the at least one processor 10 through a power management system, thereby enabling functions such as charging management, discharging management, and power consumption management through the power management system. The power supply may also include one or more DC or AC power supplies, recharging systems, power fault detection circuits, power converters or inverters, power status indicators, and other arbitrary components. The electronic device 1 may also include various sensors, Bluetooth modules, Wi-Fi modules, etc., which will not be described in detail here.

[0205] Furthermore, the electronic device 1 may also include a network interface. Optionally, the network interface may include a wired interface and / or a wireless interface (such as a Wi-Fi interface, a Bluetooth interface, etc.), which is typically used to establish communication connections between the electronic device 1 and other electronic devices.

[0206] Optionally, the electronic device 1 may further include a user interface, which may be a display, an input unit (such as a keyboard), or a standard wired or wireless interface. Optionally, in some embodiments, the display may be an LED display, a liquid crystal display, a touch-sensitive liquid crystal display, or an OLED (Organic Light-Emitting Diode) touchscreen. The display may also be appropriately referred to as a screen or display unit, used to display information processed in the electronic device 1 and to display a visual user interface.

[0207] It should be understood that the embodiments described are for illustrative purposes only and are not limited to this structure in the scope of the patent application.

[0208] The program for the extraction method of natural pigments for coatings based on microbial fermentation, stored in the memory 11 of the electronic device 1, is a combination of multiple instructions. When run in the processor 10, it can achieve the following:

[0209] Multiple initial strains were obtained, and a group of recombinant strains was obtained using the multiple initial strains and pre-constructed gene editing technology. The group of recombinant strains included multiple recombinant strains.

[0210] For each of the recombinant strains in the aforementioned recombinant strain group, a single-cell suspension was prepared to obtain a single-cell suspension set. The single-cell suspension set included multiple single-cell suspensions, and each single-cell suspension corresponded one-to-one with a recombinant strain. Multiple single-cell suspensions were subjected to fluorescence detection to obtain multiple fluorescence detection nodes. The optimal mutant strain was identified using multiple fluorescence detection nodes.

[0211] The optimal mutant strain was subjected to crude pigment extraction to obtain a crude extract of mutant pigment. The maximum absorption wavelength was determined using the crude extract of mutant pigment.

[0212] The best mutant strains were cultivated to obtain multiple best mutant strains. The multiple best mutant strains were grouped according to the preset fermentation temperature to obtain multiple mutant strain experimental groups. The absorbance of each mutant strain experimental group in the multiple mutant strain experimental groups was measured to obtain an absorbance value set. The optimal fermentation temperature was determined using the absorbance value set. The absorbance value set includes the absorbance value of each mutant strain experimental group in the multiple mutant strain experimental groups at the maximum absorption wavelength.

[0213] High-secretion fermentation broth was obtained by using the optimal fermentation temperature and the optimal mutant strain for large-scale fermentation.

[0214] The high-secretion fermentation broth was subjected to cell wall disruption to obtain a cell wall disruption suspension. A dual-enzyme synergistic pigment release operation was performed on the cell wall disruption suspension to obtain an intracellular pigment solution. The intracellular pigment solution was then microencapsulated to obtain pigment microspheres.

[0215] A verification pigment extract was extracted from the pigment microspheres. After confirming that the verification pigment extract was the preset target pigment extract, the natural pigment was obtained using the verification pigment extract.

[0216] Specifically, the processor 10's implementation method for the above instructions can be found in [reference needed]. Figures 1 to 3 The descriptions of the relevant steps in the corresponding embodiments are not repeated here.

[0217] Furthermore, if the modules / units integrated in the electronic device 1 are implemented as software functional units and sold or used as independent products, they can be stored in a computer-readable storage medium. The computer-readable storage medium can be volatile or non-volatile. For example, the computer-readable medium may include: any entity or system capable of carrying the computer program code, a recording medium, a USB flash drive, a portable hard drive, a magnetic disk, an optical disk, a computer memory, or a read-only memory (ROM).

[0218] The present invention also provides a computer-readable storage medium storing a computer program, which, when executed by a processor of an electronic device, can perform the following:

[0219] Multiple initial strains were obtained, and a group of recombinant strains was obtained using the multiple initial strains and pre-constructed gene editing technology. The group of recombinant strains included multiple recombinant strains.

[0220] For each of the recombinant strains in the aforementioned recombinant strain group, a single-cell suspension was prepared to obtain a single-cell suspension set. The single-cell suspension set included multiple single-cell suspensions, and each single-cell suspension corresponded one-to-one with a recombinant strain. Multiple single-cell suspensions were subjected to fluorescence detection to obtain multiple fluorescence detection nodes. The optimal mutant strain was identified using multiple fluorescence detection nodes.

[0221] The optimal mutant strain was subjected to crude pigment extraction to obtain a crude extract of mutant pigment. The maximum absorption wavelength was determined using the crude extract of mutant pigment.

[0222] The best mutant strains were cultivated to obtain multiple best mutant strains. The multiple best mutant strains were grouped according to the preset fermentation temperature to obtain multiple mutant strain experimental groups. The absorbance of each mutant strain experimental group in the multiple mutant strain experimental groups was measured to obtain an absorbance value set. The optimal fermentation temperature was determined using the absorbance value set. The absorbance value set includes the absorbance value of each mutant strain experimental group in the multiple mutant strain experimental groups at the maximum absorption wavelength.

[0223] High-secretion fermentation broth was obtained by using the optimal fermentation temperature and the optimal mutant strain for large-scale fermentation.

[0224] The high-secretion fermentation broth was subjected to cell wall disruption to obtain a cell wall disruption suspension. A dual-enzyme synergistic pigment release operation was performed on the cell wall disruption suspension to obtain an intracellular pigment solution. The intracellular pigment solution was then microencapsulated to obtain pigment microspheres.

[0225] A verification pigment extract was extracted from the pigment microspheres. After confirming that the verification pigment extract was the preset target pigment extract, the natural pigment was obtained using the verification pigment extract.

[0226] In the embodiments provided by this invention, it should be understood that the disclosed devices, systems, and methods can be implemented in other ways. For example, the system embodiments described above are merely illustrative, and actual implementations may have other classification methods.

[0227] The modules described as separate components may or may not be physically separate. The components shown as modules may or may not be physical units; that is, they may be located in one place or distributed across multiple network units. Some or all of the modules can be selected to achieve the purpose of this embodiment according to actual needs.

[0228] Furthermore, the functional modules in the various embodiments of the present invention can be integrated into one processing unit, or each unit can exist physically separately, or two or more units can be integrated into one unit. The integrated unit can be implemented in hardware or in the form of hardware plus software functional modules.

[0229] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the present invention.

[0230] Furthermore, it is clear that the word "comprising" does not exclude other units or steps, and the singular does not exclude the plural. Multiple units or systems stated in a system claim may also be implemented by a single unit or system through software or hardware. The term "second class" is used to indicate names and does not indicate any specific order.

[0231] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A method for extracting natural pigments for coatings based on microbial fermentation, characterized in that, The method includes: Multiple initial strains were obtained, and a group of recombinant strains was obtained using the multiple initial strains and pre-constructed gene editing technology. The group of recombinant strains included multiple recombinant strains. For each of the recombinant strains in the aforementioned recombinant strain group, a single-cell suspension was prepared to obtain a single-cell suspension set. The single-cell suspension set included multiple single-cell suspensions, and each single-cell suspension corresponded one-to-one with a recombinant strain. Multiple single-cell suspensions were subjected to fluorescence detection to obtain multiple fluorescence detection nodes. The optimal mutant strain was identified using multiple fluorescence detection nodes. The optimal mutant strain was subjected to crude pigment extraction to obtain a crude extract of mutant pigment. The maximum absorption wavelength was determined using the crude extract of mutant pigment. The best mutant strains were cultivated to obtain multiple best mutant strains. The multiple best mutant strains were grouped according to the preset fermentation temperature to obtain multiple mutant strain experimental groups. The absorbance of each mutant strain experimental group in the multiple mutant strain experimental groups was measured to obtain an absorbance value set. The optimal fermentation temperature was determined using the absorbance value set. The absorbance value set includes the absorbance value of each mutant strain experimental group in the multiple mutant strain experimental groups at the maximum absorption wavelength. High-secretion fermentation broth was obtained by using the optimal fermentation temperature and the optimal mutant strain for large-scale fermentation. The high-secretion fermentation broth was subjected to cell wall disruption to obtain a cell wall disruption suspension. A dual-enzyme synergistic pigment release operation was performed on the cell wall disruption suspension to obtain an intracellular pigment solution. The intracellular pigment solution was then microencapsulated to obtain pigment microspheres. A verification pigment extract was extracted from the pigment microspheres. After confirming that the verification pigment extract was the preset target pigment extract, the natural pigment was obtained using the verification pigment extract.

2. The method for extracting natural pigments for coatings based on microbial fermentation as described in claim 1, characterized in that, The acquisition of multiple initial strains includes: Multiple candidate strains were obtained using a pre-constructed marine microbial database; Gene sequence alignment analysis was performed on all the candidate strains, resulting in multiple classification and attribution reports; Based on the multiple classification and attribution reports, multiple target strains were identified from multiple candidate strains; Each of the multiple target strains was cultured in pre-confirmed dried seawater medium to obtain multiple stable target strains, with each target strain corresponding to a stable target strain. Intracellular pigment content was detected for each of the multiple stable target strains to obtain a set of intracellular pigment content values. The set of intracellular pigment content values ​​included multiple intracellular pigment content values, and each intracellular pigment content value corresponded one-to-one with a stable target strain. The stable target strains corresponding to intracellular pigment content values ​​that are higher than the preset intracellular pigment content threshold are identified as initial strains, resulting in multiple initial strains.

3. The method for extracting natural pigments for coatings based on microbial fermentation as described in claim 2, characterized in that, The method of obtaining a gene recombinant strain population using multiple initial strains and pre-constructed gene editing technology includes: For each of the multiple initial strains, the following procedure was performed: The initial strain was physically isolated to obtain experimental strains and control strains; Whole-genome sequencing was performed on the experimental strain to obtain its genomic information; The experimental strain was recombined using its genomic information and pre-constructed gene editing technology to obtain a primary gene-recombined strain. The primary gene recombinant strain and the control strain were fermented separately to obtain recombinant fermentation cell liquid and control fermentation cell liquid, respectively. The recombinant fermentation cell broth and the control fermentation cell broth were subjected to crude pigment extraction to obtain crude recombinant pigment extract and crude control pigment extract, respectively. Obtain the pigment mass concentration of the crude extract of recombinant pigment and the pigment mass concentration of the crude extract of control pigment; The yield increase rate was calculated using the pigment concentration of the recombinant pigment crude extract and the pigment concentration of the control pigment crude extract. The yield increase rate is compared with the preset yield increase threshold, and the primary gene recombinant strains corresponding to the yield increase rate that is greater than or equal to the yield increase threshold are identified as gene recombinant strains. Multiple recombinant strains were combined to obtain a group of recombinant strains.

4. The method for extracting natural pigments for coatings based on microbial fermentation as described in claim 3, characterized in that, The fluorescence detection of multiple single-cell suspensions yields multiple fluorescence detection nodes, including: Perform the following operation on each of the multiple single-cell suspensions: The single-cell suspension was introduced into a pre-constructed nanoreactor to obtain an initial nanoreactor; A pre-confirmed fluorescent reporter gene expression vector was added to the initial nanoreactor to obtain the labeled nanoreactor; The labeled nanoreactors were placed in a pre-confirmed constant temperature environment and cultured for 48 hours to obtain labeled microculture medium. Every 4 hours within 48 hours, the cell fluorescence density of the labeled nanoreactors was detected using a pre-confirmed high-throughput fluorescence microscope to obtain cell density sequences and fluorescence intensity sequences. Cell density-time curves were constructed using cell density sequences, and the maximum specific growth rate was obtained based on the cell density-time curves. The maximum pigment synthesis activity index was obtained based on cell density and fluorescence intensity sequences. The coefficient of variation was calculated from the fluorescence intensity sequence to obtain the coefficient of variation. The maximum specific growth rate, maximum pigment synthesis activity index, and coefficient of variation are summarized to obtain the fluorescence detection node; By summarizing the fluorescence detection nodes, multiple fluorescence detection nodes are obtained, and each fluorescence detection node corresponds one-to-one with a labeled microculture medium.

5. The method for extracting natural pigments for coatings based on microbial fermentation as described in claim 4, characterized in that, The process of identifying the optimal mutant strain using multiple fluorescence detection nodes includes: The maximum specific growth rate, maximum pigment synthesis activity index, and coefficient of variation in multiple fluorescence detection nodes were normalized to obtain multiple normalized fluorescence detection nodes. The normalized fluorescence detection nodes include the normalized maximum specific growth rate, the normalized maximum pigment synthesis activity index, and the normalized coefficient of variation. A comprehensive evaluation value is calculated using each of the multiple normalized fluorescence detection nodes and a pre-constructed comprehensive evaluation value calculation formula to obtain a comprehensive evaluation value set. The comprehensive evaluation value calculation formula is as follows: S=0.4×T max +0.5×L+0.1×(1-CV) Where S represents the comprehensive evaluation value, T max denoted as the normalized maximum specific growth rate, L represents the normalized maximum pigment synthesis activity index, and CV represents the normalized coefficient of variation. The comprehensive evaluation values ​​in the comprehensive evaluation value set are arranged in descending order to obtain a comprehensive evaluation value sequence. The top 5 comprehensive evaluation values ​​are extracted from the comprehensive evaluation value sequence. The labeled micro-culture media corresponding to the extracted 5 comprehensive evaluation values ​​are all identified as candidate micro-culture media, thus obtaining 5 candidate culture media. Verification experiments were conducted on each of the five candidate culture media, resulting in five experimental reports. Based on these reports, the optimal labeled microculture medium was determined, and the gene recombinant strain corresponding to the optimal labeled microculture medium was identified as the optimal mutant strain.

6. The method for extracting natural pigments for coatings based on microbial fermentation as described in claim 5, characterized in that, The process involves crudely extracting pigments from the optimal mutant strain to obtain a crude extract of mutant pigments. The maximum absorption wavelength is then determined using this crude extract, including: The optimal mutant strain was fermented to obtain the mutant strain fermentation broth, and the mutant strain fermentation broth was centrifuged to obtain cell precipitate; The bacterial cell precipitate was resuspended to obtain a bacterial cell suspension, and the bacterial cell suspension was then broken down to obtain a cell lysate. The cell lysate was subjected to static extraction to obtain an extraction mixture. The extraction mixture was then centrifuged to obtain a supernatant, which was used as the crude extract of pigments from the mutant strain. Absorption spectral data were obtained by scanning the crude pigment extract of the mutant strain using a pre-confirmed UV-Vis spectrophotometer across the entire wavelength range. Peak identification is performed on the absorption spectrum data to obtain the maximum absorption wavelength.

7. The method for extracting natural pigments for coatings based on microbial fermentation as described in claim 6, characterized in that, The absorbance of each mutant experimental group in the multiple mutant experimental groups was measured to obtain an absorbance value set. The optimal fermentation temperature was determined using the absorbance value set, including: Each mutant experimental group in the multiple mutant experimental groups was inoculated into a pre-confirmed fermentation medium and cultured to obtain a fermentation broth sample set, wherein the fermentation broth sample set includes multiple fermentation broth samples, and the fermentation broth samples correspond one-to-one with the mutant experimental groups. Multiple fermentation broth samples were pretreated to obtain a supernatant sample set, which included multiple supernatant samples. The absorbance of multiple supernatant samples was measured using the maximum absorption wavelength and a UV-Vis spectrophotometer to obtain a set of absorbance values. Construct a temperature-absorbance curve using the absorbance value set; The optimal fermentation temperature was determined using the temperature-absorbance curve.

8. The method for extracting natural pigments for coatings based on microbial fermentation as described in claim 7, characterized in that, The process of microencapsulating the intracellular pigment fluid to obtain pigment microspheres includes: The intracellular pigment solution and the pre-confirmed aerogel-sodium alginate composite emulsion were homogenized and emulsified at a preset homogenization speed to obtain a primary emulsion. The primary emulsion was ion-gelled to obtain multiple primary microspheres; Each primary microsphere in a plurality of primary microspheres was cross-linked and embedded in a pre-confirmed chitosan buffer to obtain a plurality of double-embedded wet microspheres; Multiple double-embedded wet microspheres were freeze-dried under vacuum to obtain multiple original pigment microspheres; Random sampling was performed on multiple original pigment microspheres to obtain a sampled original pigment microsphere set and a set of undetermined original pigment microspheres. The sampled original pigment microsphere set included multiple sampled original pigment microspheres, and the set of undetermined original pigment microspheres included multiple undetermined original pigment microspheres. The pigment embedding rate of multiple sampled original pigment microspheres was detected using a pre-confirmed pigment embedding rate detector, resulting in a set of pigment embedding rate detection values, which included multiple pigment embedding rate detection values. Multiple pigment embedding rate detection values ​​are compared with preset pigment embedding rate detection thresholds. Sampled original pigment microspheres with pigment embedding rate detection values ​​greater than or equal to the pigment embedding rate detection thresholds are identified as qualified original pigment microspheres. The qualified original pigment microspheres and sampled original pigment microspheres are used to obtain the original pigment microsphere qualification rate. The pass rate of the original pigment microspheres is compared with the preset pass rate threshold of the original pigment microspheres. If the pass rate of the original pigment microspheres is less than the threshold, the homogenization speed is adjusted and the adjusted homogenization speed is used as the preset homogenization speed. The process is repeated until the pass rate of the original pigment microspheres is greater than or equal to the threshold. The multiple undetermined original pigment microspheres are then confirmed as pigment microspheres.

9. The method for extracting natural pigments for coatings based on microbial fermentation as described in claim 8, characterized in that, After confirming that the verification pigment extract is the preset target pigment extract, the natural pigment is obtained using the verification pigment extract, including: The pigment extract was mixed with the pre-confirmed acrylic resin emulsion to obtain the paint to be tested. The pre-confirmed aluminum plate was sprayed with the color paint to be tested to obtain the sprayed aluminum plate. Color difference detection is performed on the coated aluminum plate to obtain the color difference detection value. The color difference detection value is compared with the preset color difference detection threshold. If the color difference detection value is less than the color difference detection threshold, the verification pigment extract is confirmed as the target pigment extract. The target pigment extract was purified to obtain the natural pigment.

10. A natural pigment extraction system for coatings based on microbial fermentation, characterized in that, The system includes: The mutant strain screening module is used to obtain multiple initial strains and to obtain a gene recombinant strain population using multiple initial strains and pre-constructed gene editing technology. The gene recombinant strain population includes multiple gene recombinant strains. For each of the recombinant strains in the aforementioned recombinant strain group, a single-cell suspension was prepared to obtain a single-cell suspension set. The single-cell suspension set included multiple single-cell suspensions, and each single-cell suspension corresponded one-to-one with a recombinant strain. Multiple single-cell suspensions were subjected to fluorescence detection to obtain multiple fluorescence detection nodes. The optimal mutant strain was identified using multiple fluorescence detection nodes. The fermentation condition confirmation module is used to perform crude pigment extraction on the optimal mutant strain to obtain a crude pigment extract of the mutant strain, and to confirm the maximum absorption wavelength using the crude pigment extract of the mutant strain. The best mutant strains were cultivated to obtain multiple best mutant strains. The multiple best mutant strains were grouped according to the preset fermentation temperature to obtain multiple mutant strain experimental groups. The absorbance of each mutant strain experimental group in the multiple mutant strain experimental groups was measured to obtain an absorbance value set. The optimal fermentation temperature was determined using the absorbance value set. The absorbance value set includes the absorbance value of each mutant strain experimental group in the multiple mutant strain experimental groups at the maximum absorption wavelength. The pigment microsphere synthesis module is used to carry out large-scale fermentation using the optimal fermentation temperature and the optimal mutant strain to obtain a high-secretion fermentation broth; The high-secretion fermentation broth was subjected to cell wall disruption to obtain a cell wall disruption suspension. A dual-enzyme synergistic pigment release operation was performed on the cell wall disruption suspension to obtain an intracellular pigment solution. The intracellular pigment solution was then microencapsulated to obtain pigment microspheres. The natural pigment verification module is used to extract a verification pigment extract from the pigment microspheres. After confirming that the verification pigment extract is the preset target pigment extract, the natural pigment is obtained using the verification pigment extract.

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