CN114015605B - Novel melanin-producing Rheinheimia and application thereof - Google Patents
Novel melanin-producing Rheinheimia and application thereof Download PDFInfo
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- CN114015605B CN114015605B CN202111341332.2A CN202111341332A CN114015605B CN 114015605 B CN114015605 B CN 114015605B CN 202111341332 A CN202111341332 A CN 202111341332A CN 114015605 B CN114015605 B CN 114015605B
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Abstract
The invention discloses a novel melanin-producing Rheinheimia and application thereof. The strain is Rheinheimia sp.4Y.26, which has been deposited at the microorganism strain deposit center of Guangdong province at 2021, month 6, and the deposit number is GDMCC No. 61731. The Rheinheimia 4Y26 can be used for preparing melanin. The free radical scavenging test result shows that the melanin produced by the Rheinheimia 4Y26 has excellent free radical scavenging capability and good scavenging effect on hydroxyl free radicals and DPPH free radicals, so the Rheinheimia 4Y26 has good application potential in the preparation of antioxidant products.
Description
Technical Field
The invention relates to the technical field of fermentation in microorganisms and bioengineering, in particular to novel melanin-producing Rheinheimia and application thereof.
Background
Melanin is a polymer composed of heterogeneous indole or phenol substances, is one of the most abundant natural pigments in nature, and is widely distributed in various animals, plants and microorganisms. Based on the chemical precursors synthesized, melanin is mainly composed of five classes, eumelanin, pheomelanin, neuromelanin, heteromelanin and melanin. Among them, eumelanin, pheomelanin and neuromelanin are mainly present in animal tissues, and heteromelanin and melanin are mainly present in microorganisms and plants. Eumelanin and pheomelanin synthesis substrates are typically tyrosine and phenylalanine. Tyrosine is catalyzed to form dopamine and dopaquinone. Dopamine and cysteine undergo cysteamine acylation to form cysteine-dopamine, which ultimately forms brown pigment, while dopaquinone forms eumelanin. Therefore, eumelanin contains carbon, hydrogen, oxygen and nitrogen as main elements, and brown pigment contains carbon, hydrogen, oxygen, nitrogen and sulfur as main elements. The process of melanin synthesis involves deamination and therefore contains carbon, hydrogen and oxygen as the main elements.
Melanin, a typical heterogeneous polymer, is usually bound together by covalent bonds, hydrogen bonds, and various interactions such as cation-pi, and thus its structure has high chemical complexity. Melanin is generally insoluble in acidic solutions but soluble in alkaline solutions, and therefore, a common separation and purification method for melanin is "acid precipitation and alkali dissolution". Because melanin has low solubility in most common solvents, and because its structure is highly heterogeneous, resolving the structure of melanin and understanding the relationship between structure and function is a great challenge. In addition, melanin has a maximum absorption peak under ultraviolet and visible light, and thus melanin is often used as an ultraviolet protective agent.
As an important secondary metabolite, melanin has wide application in industries such as industry, agriculture, environmental protection, medicine and the like, and is an important biological resource. Because melanin has antioxidant and radioprotective properties, melanin is often used to prepare sunscreens, anti-aging products, radioprotective materials, hair dyes, and Bt pesticides. Meanwhile, melanin can promote immune response, can resist bacteria and viruses, is also a marker in the field of medical diagnosis and treatment, and is therefore often used for disease and tumor treatment and medical imaging. Melanin produced by certain bacteria can efficiently adsorb and reduce heavy metal ions, so that heavy metal pollution in soil and water is reduced or even eliminated. Melanin also reduces the sensitivity of pathogenic bacteria to host defense systems, affects host immune responses to pathogenic bacteria, and protects the pathogenic bacteria themselves from oxidative stress by scavenging free radicals, reducing oxidative burst of host cells.
The melanin widely used at present is artificially synthesized, and has great potential safety hazard. Therefore, the biological melanin acquisition is safer, and has greater advantages in application. Natural melanin is a very wide source and can be extracted and isolated from animals, plants and microorganisms. However, compared with animals and plants, the melanin from microbial fermentation sources can compress the production period of the melanin, reduce the land and manual use cost, eliminate the influence of diseases on organisms, and remarkably improve the productivity and benefit. The current research shows that the microorganism capable of synthesizing melanin has rich classification diversity, and pseudomonas spp, streptomyces spp, vibrio spp, aeromonas spp and the like are reported. And the number of the national inventions related to these bacteria is also large. However, there is currently no related patent for melanogenesis and its use.
Disclosure of Invention
The invention provides a novel melanin-producing Rheinheimia strain, which provides a good microbial material for developing natural antioxidant products.
To achieve the above object, the present invention provides a novel melanin-producing strain, which is a strain of Rheinheimia (Rheinheimia sp.) 4Y26 deposited in the Guangdong province microorganism strain collection (GDMCC), building 5, no. 59, mitsui 100 first, xiuzhou, guangzhou City, guangdong, post code: 510070 the preservation number is GDMCC No. 61731 and the preservation date is 2021, 6 and 21.
The strain is obtained by separating from a culture water body sample of a culture plant in Taishan mountain city in the 2019 month by the inventor, and is characterized in that: gram staining was negative, single colony was round, brownish black, smooth in surface, opaque, and regular in edge. The growth temperature of the strain 4Y26 is in the range of 4-42 ℃, and the optimal growth temperature is 28 ℃; the pH range of the growth is 5-9, and the optimal growth pH is 7; the NaCl concentration range for growth is 0-1% (w/v), and the optimal NaCl concentration is 0.5% (w/v).
A second object of the present invention is to provide the use of the above-mentioned rheniheimia (rheinheimia sp.) 4Y26 for the production of melanin.
The Rheinheimia 4Y26 provided by the invention is inoculated to an R2A liquid culture medium and subjected to shaking culture at 180rpm for 7 days at 28 ℃. Taking the culture solution, centrifuging at 6000rpm for 30min, taking the supernatant, regulating the pH of the supernatant to 2, standing overnight for precipitation, centrifuging at 6000rpm for 30min, discarding the supernatant, and dissolving the precipitate by using a NaOH solution. The steps of regulating pH to precipitate melanin, centrifuging and dissolving with NaOH are repeated twice, deionized water is added for washing to neutrality, and then freeze drying is carried out. The absorbance of melanin was measured by uv-vis spectroscopy, the solubility of melanin in different solvents was tested, and the morphological characteristics and major element content of melanin were measured using a scanning electron microscope-energy spectrometer. The test results show that: the Rheinheimia 4Y26 can produce melanin, has the maximum absorption peak in the ultraviolet region, is slightly soluble in deionized water and HCl solution, can be soluble in sodium hydroxide solution and DMSO solvent, is insoluble in organic solvents such as ethanol, acetone, trichloromethane and the like, has a sheet-like microstructure, has a rough surface and holes, and contains the main elements of carbon, hydrogen and oxygen. The identification result shows that melanin produced by the Rheinheimia 4Y26 is melanin.
A third object of the present invention is to provide a viable bacterial preparation for producing melanin, which contains the above-mentioned Rheinheimia sp.) 4Y26.
A fourth object of the present invention is to provide a method for producing melanin, which comprises separating melanin from the fermentation product of Rheinheimia sp.4Y 26.
Preferably, the fermentation product comprises at least one of a fermentation broth of Rheinheimia sp.4Y26, a fermentation broth concentrate, and a fermentation broth extract.
A fifth object of the present invention is to provide the use of the above-mentioned rheniheimia (rheinheimia sp.) 4Y26 for the preparation of an antioxidant product.
The clearance rate of hydroxyl free radicals of melanin generated by the Rheinheimia 4Y26 is 95.3% at the concentration of 50mg/L, and the clearance rate of DPPH free radicals is 86.4% at the concentration of 20 mg/L. These results indicate that: the melanin produced by the Rheinheimia 4Y26 has excellent free radical scavenging capability and has good application potential in the preparation of antioxidant products.
It is a sixth object of the present invention to provide an antioxidant product comprising a fermentation product of Rheinheimia (Rheinheimia sp.) 4Y26, said fermentation product comprising at least one of a fermentation broth, a fermentation broth concentrate, a fermentation broth extract of Rheinheimia (Rheinheimia sp.) 4Y26.
Preferably, the antioxidant product is an antioxidant skin care product or a medicine.
The invention has the beneficial effects that:
the Rheinheimia sp.4Y.26 of the present invention is capable of producing and preparing melanin. The free radical scavenging test result shows that the melanin produced by the Rheinheimia 4Y26 has excellent free radical scavenging capability and good scavenging effect on hydroxyl free radicals and DPPH free radicals, so the Rheinheimia 4Y26 has good application potential in the preparation of antioxidant products.
The Rheinheimia sp. Of the present invention, 4Y26, was deposited at the Cantonese microorganism culture Collection (GDMCC) at month 21 of 2021, address: building 5, building 59, guangzhou City, guangdong, first, china, qinghai, china: 510070 with accession number GDMCC No. 61731.
Drawings
FIG. 1 shows colony morphology of the Rheinheimia 4Y26 cultured on R2A medium for 72 h.
FIG. 2 is a graph showing morphological characteristics of melanin production by Rheinheimia 4Y26.
FIG. 3 is a chart showing the ultraviolet-visible spectrum of melanin produced by the Rheinheimia species 4Y26.
FIG. 4 shows the solubility of the melanin produced by the Rheinheimia species 4Y26 in various solvents.
FIG. 5 is a morphological feature of the production of melanin by Rheinheimia 4Y26 under SEM. The left is on the 50 μm scale and the right is on the 1 μm scale.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Example 1: isolation and identification of Rheinheimia 4Y26
Collecting 500mL of a culture water sample with good water quality in a catfish culture pond in a culture factory (N21 DEG 57', 56.40', E112 DEG 50', 0.76') in the city of Taishan mountain of Jiangmen, and placing the culture water sample in a sterile sample bottle. The samples were placed in a 4 ℃ sampling box and brought back to the laboratory for subsequent processing. After the cultured water sample is brought back, the water sample is sufficiently shaken up on an ultra-clean workbench, 100 mu L of the water sample is taken and added into a 1.5mL centrifuge tube filled with 900 mu L of physiological saline, and then the water sample is diluted to 10 in sequence -2 、10 -3 、10 -4 、10 -5 And 10 -6 Suction 10 -3 、10 -4 、10 -5 And 10 -6 200. Mu.L of each diluted sample was spread on R2A medium (Qingdao sea Bo, cat# HB 0167) and cultured in a biochemical incubator at 28℃for 7 days. By visual inspection, different single colonies were picked from the medium onto a new R2A medium according to the size, color, degree of dryness, smoothness, and whether halos were generated or not, and purified by streaking multiple times until pure culture was obtained. The purified strain was numbered and was uniformly mixed with 25% glycerol (v/v) and stored in an ultra-low temperature refrigerator, thereby obtaining strain 4Y26.
The bacterial strain 4Y26 has negative gram staining, the bacterial colony morphology of the bacterial strain when being cultured on an R2A culture medium for 3 days is shown as a figure 1, and a single bacterial colony is round, has the diameter of 0.6-1.1 mm, is brownish black, has smooth surface, is opaque and has neat edges.
Example 2: 16S rRNA Gene sequence analysis and preservation of Rheinheimia 4Y26
Genomic DNA of strain 4Y26 was extracted and purified using HiPure bacterial DNA extraction kit (cat No. D3146-03, guangzhou Mei Biotechnology Co., ltd.). The bacterial 16S rRNA gene amplification universal primers 27F/1492R, namely 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-GGTTACCTTGTTACGACTT-3') were used to amplify the 16S rRNA gene sequence of strain 4Y26 using the purified genomic DNA as a template. After amplification, a proper amount of PCR products are taken for gel electrophoresis detection, and under the condition that the existence of a target band is determined, the rest PCR products are sent to Suzhou gold and other intelligent biotechnology limited company for sequencing, and the sequence of the PCR products is shown as SEQ ID NO. 1. The 16S rRNA gene sequence of strain 4Y26 was submitted to the EzBioCloud database (www.ezbiocloud.net) for sequence homology alignment. Alignment of 16S rRNA gene sequences shows: strain 4Y26 and the most recently introduced model strain Rheinheimera arenilitoris J-MS1 T The similarity of the 16S rRNA gene sequences (accession number, KJ 816861) was 99.0%. Then, strain 4Y26 and model strain R.arenilitis J-MS1 T Inoculating to R2A liquid culture medium (Qingdao sea Bo, product number HB 0167-2), culturing at 28deg.C for 3 days, and collecting appropriate amount of culture to get to Shanghai Meiji biological medicine science and technology Co., ltd. Strain 4Y26 and model strain R.aroneitis J-MS1 were calculated using the online tools Genome-to-Genome Distance Calculator 3.0.3.0 (http:// ggdc. Dsmz. De/distcalc2. Php) and ANI Calculator (www.ezbiocloud.net/tools/ANI), respectively T The digitized DNA-DNA hybridization value (dDDH) and the Average Nucleotide Identity (ANI) therebetween. Genomic alignment shows that: strain 4Y26 and model Strain R.arenilitis J-MS1 T The dDDH and ANI values between the strains were 45.7% and 91.96%, respectively, both below the DDH threshold of 70% and the ANI threshold of 95-96% for bacterial species division, indicating that strain 4Y26 is a potential new species in the genus alzheimer. Thus, the strain 4Y26 identified by the present invention is a new species and strain, namely Rheinheimia sp.4Y26, which is designated Rheinheimia sp.4Y26. Currently, the strain is deposited in the microbiological bacterial collection center (GDMCC) of Guangdong, guangzhou City, guangdong, xiuzhou Jiu No. 100 first-hand No. 59 building 5, post code: 510070 with deposit numberGDMCC No. 61731, the preservation date is 2021, 6 and 21.
Example 3: research on growth characteristics of Rheinheimia 4Y26
Rheinheimia 4Y26 was inoculated onto R2A liquid medium, placed in biochemical incubators at 4, 10, 15, 20, 28, 37, 40, 42 and 45℃respectively, and after 2 weeks of incubation, growth was observed. Rheinheimia 4Y26 was inoculated into R2A liquid medium with pH values of 3, 4, 5, 6, 7, 8, 9, 10 and 11, respectively, and placed in a biochemical incubator at 28℃for 2 weeks of stationary culture. The Rheinheimia 4Y26 is inoculated into R2A liquid culture media with mass fractions of 0, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 7.0, 10.0 and 15.0% (w/v) of artificially prepared NaCl respectively, and placed in a biochemical incubator at 28 ℃ for static culture for 2 weeks. For growth pH and NaCl test, after the culture is completed, a proper amount of culture solution is taken to determine the OD 600 The growth of the Rheinheimia 4Y26 at different pH values and NaCl concentrations was determined.
Test results show that the temperature range for the growth of the Rheinheimia 4Y26 is 4-42 ℃, and the optimal growth temperature is 28 ℃; the pH range of the growth is 5-9, and the optimal growth pH is 7; the NaCl concentration range for growth is 0-1% (w/v), and the optimal NaCl concentration is 0.5% (w/v).
Example 4: identification of melanin produced by Rheinheimia 4Y26
Rheinheimia 4Y26 was inoculated into R2A liquid medium and cultured with shaking at 180rpm at 28℃for 7 days. The culture broth was centrifuged at 6000rpm for 30min, the cells were discarded, the pH of the supernatant was adjusted to 2 with 6mol/L HCl solution, and the supernatant was allowed to stand overnight to allow melanin to precipitate sufficiently. Centrifuging at 8000rpm for 30min to obtain precipitate as crude melanin extract. The crude extract was fully dissolved with 1mol/L NaOH solution, then centrifuged at 8000rpm for 30min, the pH of the supernatant was again adjusted to 2, left overnight, centrifuged at 8000rpm for 30min to obtain a flocculent precipitate, resuspended in deionized water, washed twice to neutrality, centrifuged at 8000rpm for 10min to obtain a purified melanin extract (i.e., melanin sample), and freeze-dried.
And dissolving a proper amount of purified melanin sample in a 1mol/L NaOH solution, and carrying out ultraviolet-visible absorption spectrum analysis under the wavelength of 200-700 nm by taking a blank NaOH solution (1 mol/L) as a reference. And (3) taking a small amount of melanin samples, respectively adding the melanin samples into solutions or solvents such as deionized water, hydrochloric acid (6 mol/L), naOH (1 mol/L), ethanol, acetone, chloroform and DMSO, and fully and uniformly mixing the solutions or solvents, and observing the solubility of the solutions. A small amount of melanin sample was placed on the conductive gel, and then morphological features of melanin were observed under different magnification. An appropriate observation region is selected, and analysis of the element content in the fixed-point region is performed using a spectrometer.
The morphological characteristics of melanin produced by the Rheinheimia 4Y26 are shown in FIG. 2, and the melanin produced by the Rheinheimia 4Y26 is loose, powdery and light in weight. The ultraviolet-visible spectrum of melanin produced by the Rheinheimia 4Y26 is shown in FIG. 3, the melanin produced by the Rheinheimia 4Y26 has the maximum absorption peak in the ultraviolet region with the wavelength of 200-250 nm, and then the absorbance gradually decreases along with the increase of the wavelength, which is similar to the trend of the ultraviolet-visible absorption spectrum of the identified melanin. The solubility of melanin produced by the Rheinheimia 4Y26 in different solvents is shown in FIG. 4, the melanin produced by the Rheinheimia 4Y26 is slightly soluble in deionized water and HCl solution (6 mol/L), is easily soluble in sodium hydroxide solution (1 mol/L) and DMSO solvent, and is insoluble in organic solvents such as ethanol, acetone, chloroform and the like, which are consistent with the reported melanin solubility test results. The morphological characteristics of melanin produced by the Rheinheimia 4Y26 under SEM are shown in figure 5, and the melanin produced by the Rheinheimia 4Y26 is in a large rigid sheet shape with smooth surface on the scale of 50 μm (left in figure 5); on a 1 μm scale, the surface of melanin is rough and holes of 20 to 40nm are distributed (right in FIG. 5). Elemental analysis of the melanin samples by the spectrometer showed that: the atomic percentages of carbon, oxygen and sulfur elements in melanin produced by the Rheinheimia 4Y26 are 71.0, 28.7 and 0.3%, respectively. Melanin characterization shows that: the melanin produced by the Rheinheimia 4Y26 is melanin.
Example 5: identification of melanin-scavenging free radical produced by Rheinheimia 4Y26
Measurement of melanin produced by Rheinheimia 4Y26 by salicylic acid methodIs effective in scavenging hydroxyl radicals. 1mL 9mmol/L FeSO 4 The solution, 1mL of 9mmol/L dihydroxybenzoic acid (in 95% ethanol) solution and 1mL of melanin solution of different concentrations were mixed. 1mL of 8.8mmol/L H was added 2 O 2 The reaction was started from solution and the mixture was incubated at 37℃for 30min. All reagents are ready for use. The absorbance of the incubation mixture was measured at 510nm using deionized water as a control. The hydroxyl radical scavenging activity η (%) = [ a ] was calculated according to the following formula 0 -(A x -A x0 )]/A 0 X 100%, where A 0 Is blank control absorbance, A x Is the absorbance of the sample after adding melanin, A x0 Is free of developer H 2 O 2 Absorbance values. The test results show that: the hydroxyl radical scavenging rates of melanin produced by the Rheinheimia 4Y26 at the concentrations of 10, 20 and 30mg/L are 91.6, 94.2 and 95.3 percent respectively, which shows that the melanin produced by the Rheinheimia 4Y26 has excellent hydroxyl radical scavenging capability.
The DPPH radical scavenging ability of melanin produced by Rheinheimia 4Y26 was determined using the modified 2,2' -diphenyl-1-cumyl radical (DPPH) method. 1mL of melanin solutions of different concentrations were mixed with 3mL of 0.04mg/mL DPPH (in 95% ethanol). The mixture was kept protected from light for 30min. The absorbance of the mixture was measured at 517 nm. DPPH radical scavenging Activity ζ (%) = [1- (A) was calculated according to the following formula 0 –A S )/A 0 ]X 100%, where A 0 Is the absorbance obtained by mixing 3mL of 95% ethanol with DPPH solution, A S Is the absorbance of the DPPH solution after mixing reaction with the sample. The test results show that: the DPPH radical scavenging rates of melanin produced by the Rheinheimia 4Y26 at the concentrations of 2, 5, 10 and 20mg/L are 48.3, 64.7, 77.8 and 86.4 percent respectively, which indicates that the melanin produced by the Rheinheimia 4Y26 has excellent DPPH radical scavenging capability.
The results show that the Rheinheimia (Rheinheimia sp.) 4Y26 is a novel functional strain for producing melanin, can efficiently remove hydroxyl free radicals and DPPH free radicals, and has great application potential in the preparation of antioxidant products.
The foregoing is merely a preferred embodiment of the present invention, and it should be noted that the above-mentioned preferred embodiment should not be construed as limiting the invention, and the scope of the invention should be defined by the appended claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and such modifications and adaptations are intended to be comprehended within the scope of the invention.
Sequence listing
<110> Guangdong province microorganism institute (Guangdong province microorganism analysis and detection center)
<120> novel melanin-producing Rheinheimia and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
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<212> DNA
<213> Rheinheimia 4Y26 (Rheinheimia sp.4Y26)
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agcggcggac gggtgagtaa tgcgtaggaa gctacccgac agagggggat accagttgga 60
aacgactgtt aataccgcat aatgtctacg gaccaaagtg tgggaccttc gggccacatg 120
ctgtcggatg cgcctacgtg ggattagcta gttggtgagg taatggctca ccaaggcgac 180
gatccctagc tggtttgaga ggatgatcag ccacactgga actgagacac ggtccagact 240
cctacgggag gcagcagtgg ggaatattgg acaatgggcg caagcctgat ccagccatgc 300
cgcgtgtgtg aagaaggcct tcgggttgta aagcactttc agtagggagg aagagttgag 360
tgttaatagt actcagcttt gacgttacct acagaagaag caccggctaa ctctgtgcca 420
gcagccgcgg taatacagag ggtgcaagcg ttaatcggaa ttactgggcg taaagcgcac 480
gcaggcggtt ggttaagtca gatgtgaaag ccccgggctc aacctgggaa ttgcatttga 540
aactggccaa ctagagtacg tgagaggggg gtagaattcc aagtgtagcg gtgaaatgcg 600
tagagatttg gaggaatacc agtggcgaag gcggccccct ggcacgatac tgacgctcag 660
gtgcgaaagc gtggggagca aacaggatta gataccctgg tagtccacgc cgtaaacgat 720
gtctactagc tgttcgtggt cttgtactgt gagtagcgca gctaacgcac taagtagacc 780
gcctggggag tacggtcgca agattaaaac tcaaatgaat tgacgggggc ccgcacaagc 840
ggtggagcat gtggtttaat tcgacgcaac gcgaagaacc ttacctactc ttgacatcta 900
gcgaagattg tagagatacg attgtgcctt cgggaacgct aagacaggtg ctgcatggct 960
gtcgtcagct cgtgttgtga aatgttgggt taagtcccgc aacgagcgca acccttatcc 1020
ttagttgcca gcgagtaatg tcgggaactc tagggagact gccggtgata aaccggagga 1080
aggtggggac gacgtcaagt catcatggcc cttacgagta gggctacaca cgtgctacaa 1140
tggtatgtac agagggaggc aagctggcga cagtgagcgg atctcttaaa gcatatcgta 1200
gtccggatcg cagtctgcaa ctcgactgcg tgaagtcgga atcgctagta atcgcaaatc 1260
agaatgttgc ggtgaatacg ttcccgggcc ttgtacacac cgcccgtcac accatgggag 1320
tgggttgcaa aagaagtag 1339
Claims (7)
1. Rheinheimia sp.) 4Y26, deposited under the number: GDMCC No. 61731.
2. Use of the chlamydomonas reinhardtii (rheinheimia sp.) 4Y26 according to claim 1 for the production of melanin.
3. A viable bacterial preparation for producing melanin, characterized in that the viable bacterial preparation contains the Rheinheimia sp.4Y26 of claim 1.
4. A method for producing melanin, characterized in that melanin is isolated from the fermentation product of Rheinheimera (Rheinheimerasp) 4Y26 according to claim 1.
5. The method according to claim 4, wherein the fermentation product comprises at least one of a fermentation broth of Rheinheimia sp.) 4Y26, a fermentation broth concentrate, and a fermentation broth extract.
6. Use of the chlamydia reinhardtii (rheinheimia sp.) 4Y26 according to claim 1 for the preparation of an antioxidant skin care product or medicament.
7. An antioxidant skin care product or medicament comprising the purified melanin extract of the fermentation supernatant of the Rheinheimia sp 4Y26 of claim 1.
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