A blue pigment-producing streptomyces achromogenes and a method for producing a blue pigment

Abstract

The application belongs to the technical field of pigment production, and particularly relates to a blue pigment-producing Streptomyces discolor and a method for producing blue pigment. The application provides a blue pigment-producing Streptomyces discolor and a method for producing blue pigment. The Streptomyces discolor (Streptomyces discolor T12) provided by the application is preserved in the China General Microbiological Culture Collection Center, and has a preservation number of CGMCC No. 4.7887. The blue pigment produced by fermentation of the Streptomyces discolor is a natural pigment, has high safety, has good biological activity, and also provides a new way for industrial production of natural blue pigment, and has good application prospects in the food, dyeing and weaving, and pharmaceutical industries.

Description

Technical Field

[0001] The invention belongs to the technical field of pigment production, and particularly relates to a blue pigment-producing Streptomyces versicolor and a method for producing the blue pigment. Background Art

[0002] Pigments are natural or synthetic substances that impart a specific color to a substance. They are widely used in the food and textile industries. According to statistics, the textile industry alone consumes approximately 1.3 million tons of dyes, pigments, and dye precursors annually, with a total value exceeding US$23 billion. Based on their source, pigments are divided into natural pigments and synthetic pigments. Synthetic pigments are primarily organic pigments extracted from coal tar or chemically synthesized using aromatic compounds as raw materials. They have strong tinting power and are inexpensive. However, some synthetic pigments are difficult to degrade, polluting the environment while also harming human health. Therefore, there is an urgent need to develop environmentally friendly and safe natural pigments.

[0003] Natural pigments are abundantly available, primarily from various colored substances isolated or extracted from plants, animals, and microorganisms. These pigments can be used to maintain, enhance, or impart color to foods, pharmaceuticals, or cosmetics, finding important applications in industries such as food, textile printing and dyeing, and cosmetics. Natural pigments are widely found in a variety of organisms and are categorized as animal, plant, and microbial pigments. However, the availability of plant and animal materials is constrained by various factors, such as resources and the environment, resulting in insufficient production and high costs, limiting their application and development. The production of natural pigments through microbial fermentation overcomes many of the drawbacks of producing natural pigments from plant and animal sources and is readily industrializable, making it increasingly popular among researchers.

[0004] At present, the natural pigment reported in the document is mainly based on red and yellow pigment, blue pigment is very rare, and blue pigment is as one of three primary colors, can be deployed into multiple pigment mutually with red and yellow pigment for use, therefore the source kind that expands blue pigment is of great significance, the blue pigment that current market uses is still based on synthetic pigment, natural blue pigment is less, and many come blue vitriol ore, Polygonum indigofera, Indigofera sibiricum and Gardenia jasminoides etc., seldom have the blue pigment from microorganism source.Wherein, Gardenia jasminoides blue pigment manufacture process is complicated, cost is high, and raw materials are limited, and on large-scale industrial production, there is certain difficulty.The microorganism that can produce blue pigment of open report at present seldom, mainly contains coelicolor streptomyces (Streptomyces coelicolor) A3, pseudomonas, Duganella sp. (Duganella sp.) T2013, Duganella B2, Chromobacterium (Chromobacterium), Janssen bacillus (Janthinobacterium lividum) and Pseudomonas fluorescens (Pseudomonas fluorescens) B1 etc. The types, quantities and pigment yields of these strains are far from meeting people's needs for natural blue research and development.

[0005] Streptomyces strains belong to the Bacteria domain, Actinomycetota, Actinomycetes, Kitasatosporales, and Streptomycetaceae. Currently, more than 700 described species have been discovered. It is reported that more than 50% of the antibiotics currently used by humans are produced by Streptomyces. At the same time, strains of Streptomyces can produce a variety of bioactive substances. For example, some strains can produce various enzymes and secondary metabolites such as pigments and alkaloids, and some can repair heavy metals in the soil. Among them, Streptomyces coelicolor and Streptomyces lead-variant can produce blue pigments, but their blue pigment production is not high, and further exploration of strains that can produce blue pigments is needed.

[0006] Chinese patent application CN202410284862.5 discloses an engineered bacterium for synthesizing indigo, its construction method, and application. The engineered bacterium expresses bpsA, sfp, glnA*, and gdhA; and the engineered bacterium has glsK, proB, aceA, and ldh knocked out. By knocking out the proB and aceA genes, performing site-directed mutagenesis on the glnA gene, replacing the glsK gene with the glnA* gene, and the ldh gene with the gdhA gene, and integrating the bpsA and sfp genes, the patent application blocks the formation of byproducts, increases the flux of pyruvate to the tricarboxylic acid cycle and α-ketoglutarate to glutamate, increases the glutamine concentration in the organism, and promotes the synthesis of indigo from glutamine by indigo synthase, thereby increasing indigo production.

[0007] Chinese patent application CN202211517065.4 discloses a pigment and a method for constructing an engineered bacterial strain for its production. Comparative proteomics identified a highly expressed protein in the epidermis of the umbrella of a blue jellyfish. Sequence alignment and domain analysis revealed that the protein is similar to the blue pigment precursor of the jellyfish Rhizostoma pulmonaria, and the protein was named jellyfish blue pigment. A gene expression vector for the jellyfish blue pigment was constructed, and Rhodosporidium toruloides yeast was genetically engineered to create an industrial yeast strain that produces the blue pigment. Summary of the Invention

[0008] Current synthetic pigments have the disadvantages of being difficult to degrade, polluting the environment, and harming human health. To overcome the shortcomings of these technologies, the present invention provides a blue pigment-producing strain of Streptomyces discolor T12 and a method for producing the blue pigment. The Streptomyces discolor T12 provided by the present invention has been deposited with the General Microbiology Center of the China General Culture Collection Administration under the accession number CGMCC No. 4.7887. The blue pigment produced by fermentation with this strain is a natural pigment with high safety and excellent biological activity. It also provides a new approach for the industrial production of natural blue pigment and has promising application prospects in industries such as food, dyeing and textiles, and medicine.

[0009] The present invention achieves the above technical effects through the following technical solutions:

[0010] One of the purposes of the present invention is to provide a blue pigment-producing Streptomyces versicolor T12.

[0011] The Streptomyces versicolor T12 provided herein belongs to the family Streptomycetes and is a new species of the genus Streptomyces. This strain was deposited in June 2024 with the General Microbiology Center of the China Culture Collection Administration (address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing) under the registration number CGMCC No. 4.7887. A biological material deposit certificate for patent procedures was received from this institution on June 19, 2024, and the deposit was registered.

[0012] The growth characteristics of this strain are as follows: Streptomyces versicolor T12 is an aerobic, Gram-positive, and catalase-positive bacterium. It forms well-developed aerial hyphae on ISP2 and soil extract media. The hyphae are spiral-shaped, and the spores have external spines, resembling cockleburs. The colonies are round, convex, bluish-gray or white on the upper side, and blue on the lower side. The growth temperature range is 15-42°C. It tolerates NaCl ≤ 6% and has a pH range of 4-12. It does not produce melanin on ISP6 or ISP7. It is negative for nitrite reduction and can hydrolyze esculin, gelatin, Tween 80, and starch, but not arginine. It does not produce urease or hydrogen sulfide. It is negative for assimilating N-acetylglucosamine, capric acid, adipic acid, citric acid, and phenylacetic acid. It can utilize D-mannose, D-mannitol, D-glucose, and glycerol as sole carbon sources, but cannot utilize octanoic acid, adipic acid, malic acid, citric acid, or D-ribose as sole carbon sources. The 16S rDNA sequence of Streptomyces versicolor T12 of the present invention is shown in SEQ.No.1. In GenBank, the model strain with the highest similarity to its 16S rDNA gene sequence is S.cyaneus NRRL B-2296 T (99.6%). The G+C content of DNA was 70.2 mol%, which is consistent with the characteristics of Streptomyces. At the same time, the results of genome sequence analysis of Streptomyces versicolor T12 showed that Streptomyces versicolor T12 was similar to S.cyaneus NRRL B-2296. T The digital DNA-DNA hybridization value was 52.1%, which was far lower than 70%. The results of polyphasic taxonomic identification showed that the Streptomyces versicolor T12 was a new species of Streptomyces.

[0013] Another object of the present application is to provide an application of the above-mentioned strain, namely, the application of the above-mentioned strain in the production of blue pigment by fermentation.

[0014] Another object of the present application is to provide a method for producing blue pigment using the above-mentioned strain, wherein the method comprises fermenting and culturing Streptomyces versicolor T12, collecting the bacteria by centrifugation after the fermentation is completed, extracting the blue pigment using an organic solvent, and concentrating and drying the extract by rotary evaporation to obtain a crude blue pigment extract.

[0015] As a preferred technical scheme, the liquid culture medium for the fermentation of Streptomyces discolor T12 contains 103 g of sucrose, 10 g of glucose, 10.12 g of MgCl2·6H2O, 0.25 g of K2SO4, 5.73 g of TES, 3-5 g of yeast extract, 0.1 g of casein amino acid, 2 mL of trace elements, and distilled water is added to 1000 mL, and after sterilization, 10 mL of potassium dihydrogen phosphate (0.5%), 15 mL of L-proline (20%), 4 mL of calcium chloride (5M), and 7 mL of sodium hydroxide (1M) (to adjust pH to 7.0-7.2) are added before use.

[0016] As a preferred technical scheme, the fermentation culture condition of the Streptomyces discolor T12 is fermentation culture at 8-28℃ for 3-7 days.

[0017] As a preferred technical scheme, the shake flask fermentation culture condition of the Streptomyces discolor T12 is fermentation for 3-7 days, liquid volume is 50-100 mL / 500 mL, inoculation amount is 1-5%, pH is 7.0-7.4, temperature is 25-28℃, and shaking culture.

[0018] As a further preferred technical scheme, the organic solvent is one of ethyl acetate, tetrahydrofuran or acetone.

[0019] The present application discloses a blue pigment extraction method.

[0020] Compared with the prior art, the present application has the following beneficial effects:

[0021] 1. The Streptomyces T12 is a newly discovered new strain of Streptomyces, which is obviously different from the existing strains of the same genus. The genomic sequence analysis result of the Streptomyces T12 shows that the digital DNA-DNA hybridization value of the Streptomyces T12 and S. cyaneus NRRL B-2296T is 52.1%, which is far lower than 70%, and the polyphasic taxonomic identification result shows that the Streptomyces T12 is a new strain of Streptomyces.

[0022] 2. The pigment yield of the Streptomyces T12 is obviously higher than that of the Streptomyces coelicolor, and the blue pigment yield of the Streptomyces T12 is more than 3 times that of the blue pigment yield of the Streptomyces coelicolor M145.

[0023] 3. The pigment produced by Streptomyces T12 of the present invention has good thermal stability. When treated at 70-100°C for 2 hours, the absorbance value of its aqueous solution at 584nm remains basically unchanged. The pigment stability is significantly higher than that of pigments produced by other reported strains. The pigment loss rate is 2.55% when treated at 100°C for 2 hours; while the blue pigment produced by Pseudomonas fluorescens B1 CGMCC No.1194 loses 60.57% when treated at 100°C for 4 hours; the blue pigment produced by Xanthomonas sp. B29 CGMCC No.3699 loses 18.8% when treated at 100°C for 2 hours. Therefore, the pigment loss rate after 4 hours is greater than 18.8%.

[0024] Collection information:

[0025] Species name: Streptomyces versicolor

[0026] Latin: Streptomyces discolor

[0027] Strain number: T12.

[0028] Classification name: Streptomyces discolor T12.

[0029] Depository: General Microbiology Center of China Culture Collection Administration.

[0030] Abbreviation of the preservation agency: CGMCC.

[0031] Address of the depository institution: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0032] Deposit number: CGMCC No.4.7887.

[0033] Date of preservation: June 19, 2024. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 This is a phylogenetic tree constructed based on the 16S rRNA gene sequences of Streptomyces versicolor T12 and its reference strain.

[0035] Figure 2 This is a colony photo of Streptomyces versicolor T12 of the present invention.

[0036] Figure 3 This is an optical microscope photograph of the mycelium of Streptomyces versicolor T12 of the present invention.

[0037] Figure 4 This is a scanning electron microscope photo of the conidia and spores of Streptomyces versicolor T12 of the present invention.

[0038] Figure 5The blue pigment crude extract of Streptomyces discolor T12 and the thin layer chromatography detection results of Streptomyces coelicolor.

[0039] Figure 6 The results of the solubility characteristics of the pigment in different solvents.

[0040] Figure 7 The color change of the blue pigment solution under different pH and the full wavelength scanning absorption curve.

[0041] Figure 8 The effect of ultraviolet rays on the stability of the blue pigment crude product.

[0042] Figure 9 The effect of food additives on the stability of the blue pigment crude product.

[0043] Figure 10 The effect of metal ions on the blue pigment crude product. DETAILED DESCRIPTION

[0044] The application will be described in further detail below with reference to the accompanying drawings:

[0045] A method for producing a blue pigment of the application, the related experiments for preparing the blue pigment are as follows:

[0046] Example 1, isolation, screening and identification of the Streptomyces discolor T12 strain of the application

[0047] 1, sample collection:

[0048] The soil is collected from the Nongcuiyuan soil sample of Anhui Agricultural University in Hefei, Anhui Province.

[0049] 2, isolation and screening of the strain

[0050] Soil extract medium: take 500g of soil and add it to 2L of sterilized water, stir well, stand for 10-20min, take the supernatant, dilute it by 3 times and 10 times respectively, add 2% agar, pH 7.0-7.2, prepare the soil extract medium, pour the plate.

[0051] Isolation by dilution method: take 10.0g of soil sample and pour it into a triangular flask containing 100mL of sterile water, shake well, and then dilute it by gradient, to prepare the soil suspension; take 100μL of the soil suspension and spread it on the above separation medium plate, cultivate at 28℃ for 2-3 days, select blue single colonies, and purify them by repeated streaking, to grow vigorous blue single colonies on the soil extract medium; observe them under a microscope until they are pure strains (cell morphology is consistent), name this strain as T12, which is Streptomyces discolor T12. Store it in a 4℃ refrigerator for short-term preservation, activate it before experiment, or store it in a mixture of R2A and 40% glycerol at -80℃ for long-term preservation.

[0052] Prepare ISP2 solid culture medium for activating the strain: 0.5 g yeast extract powder, 0.5 g peptone, 0.5 g casein hydrolysate, 0.5 g glucose, 0.5 g soluble starch, 0.3 g dipotassium hydrogen phosphate, 0.024 g anhydrous magnesium sulfate, 0.3 g sodium pyruvate, 20.0 g agar, add 1000 mL water, and adjust the final pH to 7.0-7.4.

[0053] 3. Identification of strains

[0054] After 3 days of culture at 28°C in R2A medium, strain samples were collected and observed for hyphae and spore morphology under light and electron microscopy. Gram staining was performed using the standard Gram staining method. The culture characteristics of Streptomyces versicolor T12 were analyzed under different temperatures (8, 20, 28, 30, and 33°C), different NaCl concentrations (0, 0.3, 0.5, 0.7, 1, 1.5, and 2%), and different initial medium pH values ​​(5.0, 6.0, 7.0, 8.0, 9.0, and 10.0). Catalase activity was measured in freshly grown cells using a 3% (v / v) H2O2 solution. Oxidase activity was detected using the bioMérieux oxidase reagent according to standard procedures. Indole and hydrogen sulfide assays were performed according to the Tindall assay. The strain's ability to utilize different carbon and nitrogen sources, as well as the amino acid and sugar composition of its cell wall, were also investigated.

[0055] The results showed that the front of the S. discolor T12 colony on the soil extract was white or blue. The colony photos of the Streptomyces discolor T12 of the present invention are as follows: Figure 2 As shown, the colony is 3-4 mm in diameter and radiates in a circular shape. The surface is covered with dense creeping aerial hyphae ( Figure 3 ). Compared with strain S.cyaneus NRRLB-2296 T The aerial hyphae of Streptomyces versicolor T12 are more developed, thicker in diameter and more numerous. The spores are spiral-shaped, with a small number of branches, gathered into dense clusters and extended to the surrounding area. Under the scanning electron microscope, the spores are arranged in a spiral shape, with thorns on the outer surface of the spores, and the shape is similar to that of Xanthium sibiricum. The spores are about 0.9-1.1 microns long and 0.4-0.5 microns wide ( Figure 4Thin-layer chromatography and high-performance liquid chromatography (HPLC) were used to analyze the cell wall amino acids, whole-cell sugars, polar lipids, and respiratory quinones of Streptomyces versicolor T12. The results showed that the main amino acid component of the cell wall of strain T12 was LL-DAP (LL-diaminopimelic acid), and the cell wall mainly contained ribose and glucose. The main polar lipids were phosphatidylethanolamine (PE), phosphatidylinositol (PI), and diphosphatidylglycerol (DPG). The cell wall also contained phosphatidylinositol mannoside (PIM), five unidentified phospholipids (PL1-5), two unidentified amino lipids (AL1-2), and one unidentified glycolipid. These results were consistent with the composition characteristics of polar lipids in Actinomycetes, indicating that strain T12 belongs to the genus Actinomycetes.

[0056] The 16S rDNA sequence of S. discolor T12 was determined. The 16S rDNA sequence of S. discolor T12 of the present invention is shown in SEQ. No. 1, and its sequence is registered in GenBank as OQ154868. A phylogenetic analysis was performed based on the 16S rDNA gene sequences of S. discolor T12 and its closest related model strains and reference strains in GenBank. A phylogenetic tree was constructed using MEGA6 software using the neighbor-joining method, maximum likelihood method, and maximum parsimony method. Figure 1 The draft genome of the strain was determined and the results showed that the strain was similar to the reference strain S.cyaneus NRRLB-2296. T The dDDH value was 51.2%, which was much lower than 70%, indicating that strain T12 was different from S. cyaneus NRRLB-2296. T The new strain has a G+C content of 70.2 mol%.

[0057] Table 1 Differences in physiological characteristics between Streptomyces versicolor T12 and existing model strains of Streptomyces

[0058]

[0059]

[0060] Note: “+” means positive reaction, “-” means negative reaction.

[0061] Example 2 Preparation of blue pigment using S. discolor T12 strain

[0062] 1. Strain activation

[0063] Prepare solid ISP2 medium. Sterilize the prepared medium at 121°C for 20-30 minutes, pour into a plate, cool, inoculate with S. discolor T12, and incubate at 28°C for 4-7 days.

[0064] Prepare R5 medium for fermentation of the strain: 103g sucrose, 10g glucose, 10.12g magnesium chloride hexahydrate, 0.25g potassium sulfate, 5.73g TES, 5g yeast extract, 0.1g casamino acids, 2mL trace elements, and distilled water to 1000mL. For solid culture, add agar to a final concentration of 2.0%. After sterilization, add the following: 10mL potassium dihydrogen phosphate (0.5%), 15mL L-proline (20%), 4mL calcium chloride dihydrate (5M), 7mL sodium hydroxide (1M), and adjust the pH to 7.0 before use. Select an appropriate number of colonies and inoculate them into liquid medium. Incubate at 28°C with shaking for 4-7 days.

[0065] 2. Extraction of blue pigment

[0066] The pigment produced by Streptomyces versicolor T12 turns blue under alkaline conditions and red under acidic conditions. Ethyl acetate can only be extracted under acidic conditions, so ethyl acetate is the optimal solvent for pigment extraction. Dilute hydrochloric acid is added dropwise to the fermented bacterial solution. When the solution changes from blue to red, an organic solvent such as ethyl acetate is added to extract the blue pigment. The solution is shaken at 16°C for 2 hours. After standing and stratification, the upper ethyl acetate phase pigment solution is transferred to a new bottle. If pigment still remains in the lower bacterial solution, the above steps can be repeated until no more pigment is extracted. The resulting supernatant is evaporated and dried on a rotary evaporator to obtain the crude blue pigment fraction.

[0067] 3. Thin layer chromatography analysis of blue pigment crude extract

[0068] A 50×100G silica gel plate was used, and the developing solvent was benzene:acetic acid (9:1). The results were as follows: Figure 5 As shown, the blue pigment contains at least five components, while the blue pigment extract of Streptomyces coelicolor contains only two components. The blue pigment components produced by the two strains are significantly different. The blue pigment yield of Streptomyces versicolor T12 of the present invention is significantly higher than that of Streptomyces coelicolor. The blue pigment yield of Streptomyces T12 of the present invention is more than three times that of Streptomyces coelicolor.

[0069] Example 3 Detection of the physicochemical properties of the blue pigment obtained by fermentation of the S. discolor T12 strain of the present invention

[0070] 1. Solubility characteristics: Weigh 5 mg of crude blue-purple pigment extract and dissolve it in 1 mL of water, acetonitrile, methanol, ethanol, isopropanol, acetone, tetrahydrofuran, ethyl acetate, chloroform, n-hexane, petroleum ether (60-90) (the order of the above reagents is the same as Figure 6 The pigment was dissolved in each solvent in the same order as above, and then shaken and dissolved at room temperature for 1 hour. The pigment was allowed to stand and the dissolution of the pigment in each solvent was observed. The results were as follows: Figure 6 As shown, the blue pigment is soluble in acetonitrile, methanol, ethanol, acetone, tetrahydrofuran, and ethyl acetate, slightly soluble in water, and difficult to dissolve in n-hexane and petroleum ether, among which the solubility is the highest in methanol, acetone, and tetrahydrofuran.

[0071] 2. Thermal stability of pigments

[0072] The blue pigment extract obtained by the present invention was prepared into an aqueous solution with a concentration of 3 mg / mL, adjusted to pH 9, and placed in a water bath at 28°C, 40°C, 60°C, 80°C, and 100°C for 2 hours. The absorbance at 584 nm was measured and the residual pigment was calculated. The results showed that after 2 hours of high-temperature treatment, the pigment had a loss rate of less than 2%, except for a loss rate of 2.55% at 100°C, which indicated that the pigment of the present invention has strong heat resistance.

[0073] Table 2 Pigment loss rate at different temperatures

[0074] Temperature (℃) Processing time (h) Absorbance (584nm) Loss rate (%) 70 2 0.953 0.7％ 80 2 0.945 1.65％ 90 2 0.944 1.67％ 100 2 0.938 2.55％

[0075] 3. Acid and alkali stability of pigments

[0076] like Figure 7 , add HCl or NaOH solution in the pigment dissolved with ethyl hexanoate to 10mL, the pH value of solution is measured when pigment solution color changes, it is found that when pigment solution pH≥9, it is blue, solution turns blue, and with pH increase, color deepens, and during pH=7.0, pigment solution is purple, and when pH≤5.5, pigment solution is red, and full wavelength scanning is carried out using ultraviolet-visible spectrophotometer, and result shows that pigment solution has specific absorption peak at wavelength 370nm and 570nm respectively; When pigment solution pH≤5.5, this secondary peak (570nm) disappears. Moreover, the pigment obtained by the present invention and acid-base reaction discoloration belong to reversible reaction, add alkali in the pigment solution that adds acid and turns red, pigment turns blue again, adds acid again, and can become red again, and the color of this pigment solution depends on the size of pH value.

[0077] 4. Photostability of pigments

[0078] The blue pigment extract obtained by the present invention was prepared into a 3 mg / mL aqueous solution, adjusted to pH 9, added to a quartz cuvette, and irradiated under ultraviolet light. After a certain period of time, the pigment loss rate was as follows: Figure 8 The results show that the pigment solution OD 584 nm The absorbance value decreased slightly, and the pigment solution did not change significantly, indicating that it has good tolerance to ultraviolet rays.

[0079] 5. Effect of food additives on pigment stability

[0080] The blue pigment extract obtained by the present invention was prepared into a 3 mg / mL aqueous solution and adjusted to pH 9 for later use. 2.5 mL of the pigment solution was taken, 0.5 mL of 10% sucrose and 1 mL of salt (sodium chloride) solution were added respectively, and then distilled water was added to 5 mL. 2.5 mL of the pigment alcohol solution was added with 2.5 mL of deionized water as a control. After mixing and standing for a period of time, the OD values ​​of each were measured. 584 The results showed that additives such as sucrose and salt had no effect on the stability of the pigment ( Figure 9 ).

[0081] 6. Effect of metal ions on crude blue pigment

[0082] The blue pigment extract obtained by the present invention is prepared into a 3 mg / mL aqueous solution, which is adjusted to pH 9 for later use. 5 test tubes are taken and 3 mL of the pigment solution is added to each of them. A certain amount of potassium chloride, sodium chloride, magnesium chloride hexahydrate, and anhydrous aluminum chloride are added to 4 of them to make the concentration in the solution 1 mol / L. The other test tube is used as a control. After mixing and standing for a period of time, the OD values ​​of each test tube are measured. 584 The results showed that the metal ion K + 、Na + Mg 2+ and Al 3+ The addition of T12 pigment solution will increase the OD 584 The value increases, but the pigment stability is not affected much (such as Figure 10 shown).

[0083] 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 invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations that come within the meaning and range of equivalents of the claims be embraced therein.

[0084] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

[0085] The present invention is not limited to the above description of the embodiments. Any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without requiring creative work should fall within the scope of protection of the present invention.

[0086] DTD version: V1_3

[0087] File name: A blue pigment-producing Streptomyces versicolor and a method for producing blue pigment.xml

[0088] Software Name: WIPOSequence

[0089] Software version: 2.3.0

[0090] Date of Generation: 2024-07-01

[0091] Basic Information:

[0092] Current application / applicant file name: Anhui Agricultural University

[0093] Applicant's name: Anhui Agricultural University

[0094] Applicant's name / language:zh

[0095] Applicant's name or title / Latin name: Anhui Agricultural University

[0096] Inventor Name: Mei Fengmin

[0097] Inventor Name / Language:zh

[0098] Inventor Name / Latin Name:Mei fengmin

[0099] Invention Title: A blue pigment-producing Streptomyces versicolor and a method for producing blue pigment (zh)

[0100] Total number of sequences: 1

[0101] sequence:

[0102] Serial number (ID): 1

[0103] Length:1492

[0104] Molecule type: DNA

[0105] Feature Position / Qualifier:

[0106] -source,1..1492

[0107] >mol_type,genomic DNA

[0108] >organism,unidentified

[0109] Residue:

[0110]

[0111]

Claims

1. A blue pigment-producing Streptomyces chromophore ( Streptomyces discolor )T12, the depository institution is the General Microbiology Center of China Culture Collection Administration of Microorganisms, and the deposit registration number is CGMCC No.4.7887.

2. Application of Streptomyces versicolor T12 according to claim 1 in producing blue pigment by fermentation.

3. A method for producing blue pigment using the Streptomyces versicolor T12 according to claim 1, characterized in that: The Streptomyces versicolor T12 was fermented and cultured, and the cells were collected by centrifugation after the fermentation was completed. The blue pigment was extracted with ethyl acetate under acidic conditions, and the extract was concentrated and dried by rotary evaporation to obtain a crude blue pigment extract.

4. The method for producing blue pigment by utilizing the Streptomyces versicolor T12 according to claim 1 according to claim 3, characterized in that The liquid culture medium used for fermentation of Streptomyces versicolor T12 contains the following per liter: sucrose 103 g, glucose 10 g, MgCl2·6H2O 10.12 g, K2SO4 0.25 g, TES 5.73 g, yeast extract 3-5 g, casamino acids 0.1 g, trace elements 2 mL, and distilled water to 1000 mL. After sterilization, 10 mL of 0.5% potassium dihydrogen phosphate solution, 15 mL of 20% L-proline, 4 mL of 5 M calcium chloride dihydrate, and 7 mL of 1 M sodium hydroxide should be added before use.

5. The method for producing blue pigment by utilizing the Streptomyces versicolor T12 according to claim 1 according to claim 3, characterized in that The fermentation culture conditions of the Streptomyces versicolor T12 are as follows: fermentation culture at 28° C. for 3-8 days.

6. The method for producing blue pigment by utilizing the Streptomyces versicolor T12 according to claim 1 according to claim 5, characterized in that The fermentation culture conditions of the Streptomyces versicolor T12 are as follows: fermentation culture at 28° C. for 6 days.

7. The method for producing blue pigment by utilizing the Streptomyces versicolor T12 according to claim 1 according to claim 3, characterized in that When the pH of the solution containing the blue pigment is ≥9, the pigment solution is blue, and when the pH is ≤5.5, the pigment solution is red.

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