Method for preparing ultraviolet absorbent by utilizing Kocuria rhizophila and application
By isolating Rhizophora oryzae from coral mucus to prepare 3-butyl-1-isobenzofuranone UV absorber, the problems of skin allergy and environmental impact of traditional UV absorbers are solved, and highly efficient UVA band ultraviolet protection is achieved.
Patent Information
- Application Number
- CN202511910163.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-17
- Publication Date
- 2026-03-17
AI Technical Summary
Traditional UV absorbers pose risks of skin allergies and environmental impacts, and their absorption capacity in the UVA band is insufficient, making them ineffective in preventing skin tanning.
Kocuria rhizophila was isolated and screened from the mucus of corals in the South China Sea. 3-Butyl-1-isobenzofuranone was prepared as a UV absorber. The highly efficient UV absorber was obtained through a specific culture medium and extraction separation process.
The prepared 3-butyl-1-isobenzofuranone has strong UV absorption capacity in the UVA band, is environmentally friendly, has high safety, and effectively prevents skin tanning.
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Figure CN121674283A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of biotechnology, and in particular to a method and application for preparing ultraviolet absorbers using Kocuria rhizophila. Background Technology
[0002] Traditional sunscreens use UV absorbers such as benzophenone and cinnamate, which pose risks of skin allergies or cancer. Furthermore, their large-scale use ultimately leads to their release into oceans and lakes, causing adverse effects on the environment and wildlife. In addition, over 98% of UVA ultraviolet rays with wavelengths of 320–400 nm can penetrate the ozone layer and clouds to reach the Earth's surface, directly penetrating the dermis and damaging elastic and collagen fibers, causing sunburn and posing a significant health hazard. However, currently used traditional UV absorbers have insufficient absorption capacity in the UVA band.
[0003] Coral reefs are widely distributed in shallow waters between 30 degrees north and south latitude. Exposed to high levels of ultraviolet radiation, coral mucus is rich in UV-resistant substances, thus protecting the coral host from UV damage. A diverse range of microorganisms exist in coral mucus, utilizing the organic nutrients to produce UV-resistant compounds, further protecting the coral host from UV damage. Summary of the Invention
[0004] This invention utilizes a natural, UV-resistant microorganism, *Kocuria rhizophila*, isolated and screened from coral mucus in the South China Sea. The UV absorber prepared from this *Kocuria rhizophila* is 3-butyl-1-isobenzofuranone (3-Butylidenephthalide). Compared to benzophenone, a commonly used UV absorber in traditional sunscreens, it exhibits stronger UV absorption, especially in the UVA band (320-400 nm), thus effectively preventing skin tanning. Furthermore, these UV absorbers, derived from coral mucus microorganisms, possess better environmental and biocompatibility than traditionally used chemically synthesized UV absorbers, making them safer to use.
[0005] This invention is specifically implemented through the following scheme:
[0006] On the one hand, the present invention provides a natural Kocuria rhizophila BB045, which is now deposited at the China Center for Type Culture Collection, accession number CCTCC M 20252319, with a deposit date of October 22, 2025.
[0007] Secondly, the present invention provides a method for preparing an ultraviolet absorber, comprising the following steps:
[0008] (1) The rhizotrophic cocci were added to 2216E medium and cultured to obtain seed culture;
[0009] (2) The seed culture was inoculated into the bacterial fermentation medium and cultured to obtain the fermentation broth;
[0010] (3) Add ethyl acetate to the fermentation broth for extraction to obtain an extract. Then, evaporate the extract under reduced pressure to obtain a crude extract.
[0011] (4) The crude extract was extracted, separated and purified to obtain 3-butyl-1-isobenzofuranone, which is the ultraviolet absorber.
[0012] Furthermore, the method for preparing the ultraviolet absorber is as follows:
[0013] Pretreatment of crude extract: Dissolve the crude extract obtained in step (3) in 50-60 mL of methanol, add 20-30 g of 100-200 mesh silica powder and evaporate under reduced pressure to obtain the embedded crude extract;
[0014] Forward silica column chromatography: A forward silica column is packed with 100-200 mesh silica powder and 300-400 mesh silica powder. The embedded crude extract is loaded onto the column, and the following gradient elution is performed sequentially using a mixture of petroleum ether and ethyl acetate, and a mixture of dichloromethane and methanol as the mobile phase.
[0015] From 0 to 25 minutes, the eluent 1 was collected by using a mixture of petroleum ether and ethyl acetate at a volume ratio of 0:1 as the mobile phase.
[0016] For 25-50 minutes, the eluent 2 was collected by mixing petroleum ether and ethyl acetate in a volume ratio of 1:4 as the mobile phase.
[0017] For 50-75 minutes, a mixture of petroleum ether and ethyl acetate at a volume ratio of 2:3 was used as the mobile phase, and the eluent 3 was collected.
[0018] For 75-100 min, use a mixture of petroleum ether and ethyl acetate in a volume ratio of 3:2 as the mobile phase, and collect the eluent 4.
[0019] 125~150 min, using a mixture of petroleum ether and ethyl acetate at a volume ratio of 4:1 as the mobile phase, and collect the eluent 5;
[0020] For 150-175 min, a mixture of petroleum ether and ethyl acetate at a volume ratio of 1:0 was used as the mobile phase, and the eluent 6 was collected.
[0021] The eluent 7 was collected after 175-200 min of a mixture of dichloromethane and methanol at a volume ratio of 0:1 as the mobile phase.
[0022] For 200-225 minutes, a mixture of dichloromethane and methanol at a volume ratio of 1:3 was used as the mobile phase, and the eluent 8 was collected.
[0023] The eluent was collected at a volume ratio of 1:1 (dichloromethane and methanol) for 225-250 min.
[0024] Thin-layer chromatography analysis and component combination: Spot eluents 1-9 onto a thin-layer chromatography silica gel plate for thin-layer chromatography analysis. Combine eluents 1 and 2, 4 and 5, and 8 and 9, and evaporate to dryness under reduced pressure to obtain a mixture.
[0025] Add the mixture to ethanol to obtain a solution of the mixture;
[0026] The mixture solution was added to the chromatographic column and eluted with mobile phase 10 at a flow rate of 0.4 mL / min. The eluent retained for 27–29 min was collected and denoted as Fr3. Mobile phase 10 was prepared by mixing an aqueous solution of 0.05% formic acid and an acetonitrile solution containing 0.05% formic acid at a volume ratio of 1:9.
[0027] After distilling the eluent Fr3 under reduced pressure, 4 mL of methanol was added to dissolve it. The supernatant was collected by centrifugation and added to the chromatographic column. Mobile phase 11 was used for elution at a flow rate of 0.4 mL / min. The eluent retained for 14.5~15.5 min was collected, which was 3-butyl-1-isobenzofuranone. Mobile phase 2 was obtained by mixing 0.1% formic acid aqueous solution and acetonitrile in a volume ratio of 7:2.
[0028] Furthermore, the method for preparing the ultraviolet absorber is characterized in that the chromatographic column is an Agient EclipseXDB-CI8 with dimensions of 9.4 x 250 mm and a particle size of 5 μm.
[0029] Furthermore, the method for preparing the ultraviolet absorber, wherein the 2216E culture medium formula is as follows: peptone 5.0 g / L, yeast extract 1.0 g / L, ferric citrate 0.1 g / L, sodium chloride 19.45 g / L, magnesium chloride 5.98 g / L, sodium sulfate 3.24 g / L, calcium chloride 1.8 g / L, potassium chloride 0.55 g / L, sodium carbonate 0.16 g / L, potassium bromide 0.08 g / L, strontium chloride 0.034 g / L, boric acid 0.022 g / L, sodium silicate 0.004 g / L, sodium fluoride 0.0024 g / L, sodium nitrate 0.0016 g / L, disodium hydrogen phosphate 0.008 g / L, pH 7.4~7.8.
[0030] Furthermore, in the preparation method of the ultraviolet absorber, the bacterial fermentation culture medium formula is as follows: 10g tryptone, 5g yeast extract, 10g sodium chloride, and 1L sterile artificial seawater, wherein the artificial seawater formula is: NaCl: 132.6g, MgCl2·6H2O: 26.14g, MgSO4: 16.53g, CaCl2: 5.71g, KCl: 3.63g, NaHCO3: 1.01g, NaBr: 0.42g, distilled water: 5000mL, pH 7.0-7.2.
[0031] Thirdly, the present invention provides an ultraviolet absorber, characterized in that it comprises 3-butyl-1-isobenzofuranone.
[0032] Fourthly, the present invention provides the application of the aforementioned rhizotrophic coccidia in the preparation of UV-resistant products.
[0033] Fifthly, the present invention provides an application of the 3-butyl-1-isobenzofuranone as an ultraviolet absorber.
[0034] Furthermore, the application is characterized in that 3-butyl-1-isobenzofuranone absorbs ultraviolet light with a wavelength of 200~400nm. Attached Figure Description
[0035] Figure 1 Survival curves of naturally occurring *Cockia rhizophila* isolated from coral mucus under ultraviolet light.
[0036] Figure 2 DAD detection peak diagram of 3-Butylidenephthalide
[0037] Figure 3 It is 3-Butylidenephthalide. 13 C NMR spectrum
[0038] Figure 4 It is 3-Butylidenephthalide. 1 H NMR image
[0039] Figure 5 The structural formula is 3-butyl-1-isobenzofuranone (3-Butylidenephthalide).
[0040] Figure 6 UV absorption spectrum of 3-Butylidenephthalide
[0041] Figure 7 UV absorption spectrum of benzophenone (control) Detailed Implementation
[0042] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the scope of this application. Reagents not specifically described in detail herein are all conventional reagents and are commercially available; methods not specifically described in detail are all conventional experimental methods and can be learned from the prior art.
[0043] It should be noted that the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion, for example, a process, method, system, product, or device that includes a series of steps or units is not necessarily limited to those steps or units that are explicitly listed, but may include other steps or units that are not explicitly listed or that are inherent to such process, method, product, or device.
[0044] To better understand the invention and not to limit its scope, all figures indicating amounts, percentages, and other numerical values used in this application should, in all cases, be understood to be modified by the word "approximately." Therefore, unless specifically stated otherwise, the numerical parameters listed in the specification and appended claims are approximate values and may vary depending on the desired properties being sought. Each numerical parameter should at least be considered as obtained based on reported significant figures and through conventional rounding methods.
[0045] Example 1: Isolation and Identification of Microorganisms from Coral Slime
[0046] Coral was collected from the coral reef of Xidao Island in Sanya. The coral was washed with sterile artificial seawater, and the mucus was extracted using a sterile pipette and stored in centrifuge tubes. Sterile artificial seawater was then added and mixed to dilute the coral mucus.
[0047] Add 20 mL of 2216E medium to a 50 mL centrifuge tube. The medium has the following composition: peptone 5.0 g / L, yeast extract 1.0 g / L, ferric citrate 0.1 g / L, sodium chloride 19.45 g / L, magnesium chloride 5.98 g / L, sodium sulfate 3.24 g / L, calcium chloride 1.8 g / L, potassium chloride 0.55 g / L, sodium carbonate 0.16 g / L, potassium bromide 0.08 g / L, strontium chloride 0.034 g / L, boric acid 0.022 g / L, sodium silicate 0.004 g / L, sodium fluoride 0.0024 g / L, sodium nitrate (or ammonium nitrate) 0.0016 g / L, disodium hydrogen phosphate 0.008 g / L, pH 7.6±0.2 (25 ℃). Add 2 μL of coral mucus and place the tube in a constant temperature shaker at 28 ℃ and 220 rpm for enrichment culture.
[0048] Five culture media were prepared by adding artificial seawater (Table 1) and 1.8% agar: (No. 1: 22 g soybean meal, 10 g yeast extract, 5 g glucose, 2 g NaCl, 2 g MgSO4·7H2O2, 2.5 g K2HPO4, 1 L sterile artificial seawater; No. 2: 10 g tryptone, 5 g yeast extract, 10 g sodium chloride, 1 L sterile artificial seawater; No. 3: 17 g tryptone, 3 g soybean meal papain digest, 5 g NaCl, 2.5 g K2HPO4, 2.5 g glucose, 1 L sterile artificial seawater; No. 4: 10 g soluble starch, 10 g peptone, 0.5 g MgSO4·7H2O, 0.5 g K2HPO4, 1 L sterile artificial seawater; No. 5: 10 g milk powder, 5 g tryptone, 10 g glucose, 2.5 g yeast extract). g. Take 1 L of sterile artificial seawater and spread 45 μL of the above enrichment culture solution onto the corresponding solid medium, with three replicates for each medium. Incubate the plates upside down in a 28 ℃ incubator for 7-14 days. Once colonies have grown on the plates, select single colonies based on morphological characteristics for streak plating for further purification.
[0049] Table 1. Artificial Seawater Formulation
[0050]
[0051] In a clean bench, use a sterile pipette tip to pick up a single colony from the purification plate and inoculate it into a 50 mL centrifuge tube containing 20 mL of the liquid culture medium corresponding to the purification plate. Incubate the culture in a constant temperature shaker at 28 ℃ and 220 rpm for 3-4 days.
[0052] After the incubation period, add 1 μL of bacterial culture to a PCR tube in a clean bench. Prepare the PCR solution according to the PCR system preparation method in Table 2, mix thoroughly with a pipette tip, and place the PCR tube in a PCR instrument. Perform PCR amplification of 16S rDNA using universal primers (e.g., 27F and 1492R). Perform electrophoresis with 3 μL of PCR solution. Observe the DNA bands under a UV transilluminator. Cut the bands and place them in a centrifuge tube for sequencing. Compare the results with the NCBI database to determine the strain species information and complete the strain identification.
[0053] Table 2. PCR system for 16S rDNA
[0054]
[0055] Example 2: Screening of naturally occurring rhizophilic Coccidia with UV resistance
[0056] Five groups of bacterial suspensions obtained from the five culture media in Example 1 were diluted and spread onto corresponding agar plates. *E. coli* was used as a negative control. The plates were irradiated with different intensities of ultraviolet light for 0 s, 15 s, 30 s, 45 s, and 60 s. After ultraviolet irradiation, the plates were sealed with aluminum foil and incubated at 28 ℃ in the dark for 4-5 days. Once the colony count on the plates stabilized, the plates were removed, and the number of single colonies on each plate was recorded. The survival rate of the tested strains under the corresponding ultraviolet intensities was calculated using the following formula:
[0057] D = N / N0 × 100%
[0058] Where D represents the survival rate (Survival Donation).
[0059] N represents the number of bacterial strains that survive under ultraviolet irradiation;
[0060] N0 represents the number of strains that were not exposed to ultraviolet light.
[0061] By plotting a survival rate-UV intensity curve, the UV intensity (D10) corresponding to a survival rate of 10% was used as the criterion for judging the strength of the strain's UV resistance, thus screening out natural strains with UV tolerance. Following this screening process, a *Cochliobolus rhizophilus* strain with strong survival ability under UV irradiation was obtained, with a D10 of 135.30 J·m⁻¹. -2 It is 93 times that of E. coli. Figure 1 The culture medium used was No. 2, which consisted of 10 g tryptone, 5 g yeast extract, 10 g sodium chloride, and 1 L sterile artificial seawater. The strain was deposited at the China Center for Type Culture Collection on October 22, 2025, at Wuhan University, Wuhan, China.
[0062] Example 3: Isolation and purification of UV-absorbing compounds from Rhizotrophic coccidia
[0063] The *Coprinovia rumenophila* culture was inoculated into No. 2, 2216E medium and cultured in a shaker at 28 °C and 220 rpm for 48 h to obtain the seed culture. The culture medium (10 g tryptone, 5 g yeast extract, 10 g sodium chloride, 1 L sterile artificial seawater) was cultured in a shaker at 28 °C and 220 rpm for 7 days. Metabolites from the fermentation broth were extracted with ethyl acetate (3 v / v). The extract was then evaporated under reduced pressure to remove the solvent ethyl acetate, yielding a crude extract. The crude extract was dissolved in methanol, and 20 g of 100-200 mesh silica gel powder was added, followed by reduced pressure evaporation to obtain the embedded crude extract.
[0064] Pretreatment of crude extract: Dissolve the crude extract obtained in the previous step in 50 mL of methanol, add 20 g of 100-200 mesh silica powder and evaporate under reduced pressure to obtain the embedded crude extract;
[0065] Forward silica column chromatography: A forward silica column is packed with 100-200 mesh silica powder and 300-400 mesh silica powder. The embedded crude extract is loaded onto the column, and the following gradient elution is performed sequentially using a mixture of petroleum ether and ethyl acetate, and a mixture of dichloromethane and methanol as the mobile phase.
[0066] From 0 to 25 minutes, the eluent 1 was collected by using a mixture of petroleum ether and ethyl acetate at a volume ratio of 0:1 as the mobile phase.
[0067] For 25-50 minutes, the eluent 2 was collected by mixing petroleum ether and ethyl acetate in a volume ratio of 1:4 as the mobile phase.
[0068] For 50-75 minutes, a mixture of petroleum ether and ethyl acetate at a volume ratio of 2:3 was used as the mobile phase, and the eluent 3 was collected.
[0069] For 75-100 min, use a mixture of petroleum ether and ethyl acetate in a volume ratio of 3:2 as the mobile phase, and collect the eluent 4.
[0070] 125~150 min, using a mixture of petroleum ether and ethyl acetate at a volume ratio of 4:1 as the mobile phase, and collect the eluent 5;
[0071] For 150-175 min, a mixture of petroleum ether and ethyl acetate at a volume ratio of 1:0 was used as the mobile phase, and the eluent 6 was collected.
[0072] The eluent 7 was collected after 175-200 min of a mixture of dichloromethane and methanol at a volume ratio of 0:1 as the mobile phase.
[0073] For 200-225 minutes, a mixture of dichloromethane and methanol at a volume ratio of 1:3 was used as the mobile phase, and the eluent 8 was collected.
[0074] The eluent was collected at a volume ratio of 1:1 (dichloromethane and methanol) for 225-250 min.
[0075] Thin-layer chromatography analysis and component combination: Eluents 1 and 2, 4 and 5, 8 and 9 were combined, evaporated to dryness under reduced pressure to obtain a mixture; the mixture was then added to ethanol to obtain a mixture solution.
[0076] Compounds exhibiting ultraviolet absorption were traced using HPLC: The HPLC instrument was an Agilent Technologies (China) Co., Ltd. 1260 Infinity II series; the column was an Agient Eclipse XDB-CI8, 9.4 x 250 mm, with a particle size of 5 μm; the detector was a diode array detector, detecting short-wave ultraviolet light at 254 nm, medium-wave ultraviolet light at 310 nm, and long-wave ultraviolet light at 340 nm. The mobile phase consisted of an aqueous solution containing 0.05% formic acid and an acetonitrile solution containing 0.05% formic acid. The injection volume was 10 µL, and the column temperature was 30 °C. The mixture was added to the column, eluted, and the following eluent was collected:
[0077] The retention time was 27-29 min. The elution was carried out using a mixture of 0.05% formic acid aqueous solution and acetonitrile containing 0.05% formic acid at a volume ratio of 1:9 as the mobile phase. The flow rate was 0.4 mL / min, and the collected eluent was recorded as Fr3.
[0078] The eluent Fr3 from the previous step was dissolved in 4 mL of methanol after vacuum distillation. The supernatant was collected by centrifugation and injected at a volume of 50 µL into an Agilent 1260 Infinity II series HPLC semi-preparative system. The chromatographic column was Agilent Eclipse XDB-C18, with dimensions of 9.4 × 250 mm and a particle size of 5 μm. The detector was a diode array detector with a detection wavelength of 300 nm. Each peak was collected using an automatic fraction collector.
[0079] In this process, phase A is a 0.1% formic acid aqueous solution with a volume ratio of 7:2 to phase B acetonitrile, and the retention time is 14.5~15.5 min, which is 3-butyl-1-isobenzofuranone (e.g. Figure 2 (as shown)
[0080] Example 4: Structural Identification of Ultraviolet Absorbing Compounds from Rhizotrophic Cochlea
[0081] The pure compound with UV absorption prepared above was dissolved in methanol to prepare a 0.1 mg / mL solution. 500 μL of the methanol solution was transferred to a 1.5 mL centrifuge tube and centrifuged at 12000 rpm for 5 min. 50 μL of the supernatant was transferred to a LC-MS vial with an inner tube, and 1 μL was injected for LC-MS analysis. The Thermo Fisher Scientific Vanquish UPLC / Q Exactive plus mass spectrometer was used. LC-MS / MS analysis was performed on a Q Exactive Plus UHPLC system equipped with an ESI ion source. The chromatographic column was an Angilent EC-C18, 2.1 × 50 mm, with a particle size of 1.9 μm. The mobile phase consisted of an aqueous solution of 0.05% formic acid (phase A) and acetonitrile (phase B). The injection volume was 1 μL, and the column temperature was 30℃. The ESI ion source parameters are set as follows: capillary temperature 320 ℃, nozzle voltage 3.2 kV, lens RF voltage 50 V, and positive ion mode.
[0082] Mass spectrometry data of the compounds were acquired using a Thermo Fisher Xcalibur 3.0 spectrometer to obtain their molecular weights. After acquisition, the raw data were converted to .mzXML format using MSConvertGUI 3.0 software, connected to the GNPS molecular network (https: / / gnps.ucsd.edu / ), and the MS / MS results were compared with the molecular network database to observe for compounds with high matching degrees.
[0083] The structure was further identified using nuclear magnetic resonance (NMR) spectroscopy (C-H). Deuterated methanol was used as the solvent for compounds with higher polarity, while deuterated chloroform was used for compounds with lower polarity. Small, repeated applications of 500 μL of deuterated solvent were added to the sample vial, and the dissolved sample was transferred to the NMR tube until the liquid level reached 3-4 cm. The compounds were then analyzed using a Bruker Avance 600 MHz III spectrophotometer. 1 H NMR and 13 C NMR analysis, results are as follows Figure 3-4 As shown. The spectrum was analyzed using MestReNova software to determine the compound's structural formula, as shown below. Figure 5 As shown.
[0084] Example 5 Evaluation of the ultraviolet absorption capacity of compounds
[0085] The compound was completely dissolved in an appropriate amount of methanol to achieve a concentration of 1 mM. 0.364 g of benzophenone was weighed and added to 2 mL of methanol.
[0086] A 1 mM methanol solution was prepared as a positive control, and a pure methanol solution was used as a negative control.
[0087] Pipette 500 μL of each compound solution into a centrifuge tube and centrifuge at 12000 rpm for 5 min. Transfer 50 μL of the supernatant to a LC-MS vial with an inner tube. Perform HPLC full-wavelength (190-400 nm) UV absorption analysis of the compounds using a 10 μL injection volume (under the same conditions as above). Determine the maximum absorption wavelength of each compound by analyzing its UV absorption spectrum. Compare the absorbance of each compound with that of benzophenone in the UVA and UVB bands using a microplate reader. Add 280 μL of each 1 mM test solution to a 96-well plate, select a wavelength of 280-400 nm and a bandwidth of 2 nm, and read the data three times per well, taking the average value.
[0088] One compound with strong ultraviolet absorption that was isolated and identified was 3-butyl-1-isobenzofuranone (3-Butylidenephthalide). Figure 6 As shown (where the vertical axis represents the response value, with an upper limit of 100 indicating an ultraviolet absorption intensity of 3000 mAU at the corresponding wavelength, and the horizontal axis represents the wavelength), this compound exhibits very strong ultraviolet absorption capabilities. Compared to traditional ultraviolet absorbers such as benzophenone (e.g., Figure 7 As shown in Table 3, this compound has a stronger UV absorption capacity than benzophenone, especially in the UVA band (320-400 nm) where benzophenone has weak absorption. 3-Butyl-1-isobenzofuranone has very strong UV absorption (Table 3).
[0089] Table 3. UV absorbance of 3-Butylidenephthalide
[0090]
[0091] In summary, the above description is merely a preferred embodiment of this application, but the scope of protection of this application is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope provided in this application should be included within the scope of protection of this application.
Claims
1. A Kocuria rhizophila BB045, which is deposited in China General Microbiological Culture Collection Center with the accession number CCTCC M 20252319. 2.A method for preparing an ultraviolet absorber, comprising the following steps: (1) adding the Kocuria rhizophila of claim 1 into 2216E medium to obtain a seed liquid; (2) inoculating the seed liquid into a bacterial fermentation medium to obtain a fermentation liquid; (3) adding ethyl acetate into the fermentation liquid to obtain an extraction liquid, and then performing rotary evaporation under reduced pressure to obtain a crude extract; (4) separating and purifying the crude extract to obtain 3-butyl-1-isobenzofuranone, which is the ultraviolet absorber. 3.The method for preparing the ultraviolet absorber of claim 2, comprising the following steps: (1) crude extract pretreatment: dissolving the crude extract obtained in step (3) of claim 2 with 50-60 mL of methanol, adding 20-30 g of 100-200 mesh silica gel powder, and then performing rotary evaporation under reduced pressure to obtain the embedded crude extract; (2) forward silica gel column chromatography separation: filling a forward silica gel column with 100-200 mesh silica gel powder and 300-400 mesh silica gel powder, and then loading the embedded crude extract, using a mixture of petroleum ether and ethyl acetate, a mixture of dichloromethane and methanol as the mobile phase, and performing the following gradient elution; 0-25 min, using a mixture of petroleum ether and ethyl acetate with a volume ratio of 0:1 as the mobile phase 1 to collect eluate 1; 25-50 min, using a mixture of petroleum ether and ethyl acetate with a volume ratio of 1:4 as the mobile phase 2 to collect eluate 2; 50-75 min, using a mixture of petroleum ether and ethyl acetate with a volume ratio of 2:3 as the mobile phase 3 to collect eluate 3; 75-100 min, using a mixture of petroleum ether and ethyl acetate with a volume ratio of 3:2 as the mobile phase 4 to collect eluate 4; 125-150 min, using a mixture of petroleum ether and ethyl acetate with a volume ratio of 4:1 as the mobile phase 5 to collect eluate 5; 150-175 min, using a mixture of petroleum ether and ethyl acetate with a volume ratio of 1:0 as the mobile phase 6 to collect eluate 6; 175-200 min, using a mixture of dichloromethane and methanol with a volume ratio of 0:1 as the mobile phase 7 to collect eluate 7; 200-225 min, using a mixture of dichloromethane and methanol with a volume ratio of 1:3 as the mobile phase 8 to collect eluate 8; 225-250 min, using a mixture of dichloromethane and methanol with a volume ratio of 1:1 as the mobile phase 9 to collect eluate 9; (3) thin layer chromatography analysis and component combination: combining eluate 1 and 2, 4 and 5, 8 and 9, and then performing rotary evaporation under reduced pressure to obtain a mixture; (4) adding the mixture into ethanol to obtain a mixture solution; The mixture solution is added to the chromatographic column, eluted with mobile phase 10 at a flow rate of 0.4 mL / min, and the eluate with a retention time of 27-29 min is collected as Fr3; the mobile phase 10 is obtained by mixing 0.05% formic acid aqueous solution and 0.05% formic acid acetonitrile solution at a volume ratio of 1:9; (5) The eluate Fr3 is evaporated under reduced pressure, then 4 mL of methanol is added for dissolution, centrifuged, and the supernatant is added to the chromatographic column, eluted with mobile phase 11 at a flow rate of 0.4 mL / min, and the eluate with a retention time of 14.5-15.5 min is collected as 3-butyl-1-isobenzofuranone; the mobile phase 2 is obtained by mixing 0.1% formic acid aqueous solution and acetonitrile at a volume ratio of 7:
2.
4. The method for preparing the ultraviolet absorber as described in claim 3, characterized in that, The chromatographic column is Agient EclipseXDB-CI8, with a size of 9.4x250 mm and a particle size of 5 μm.
5. The preparation method of the ultraviolet absorber according to claim 2, wherein the culture medium formula is as follows: proteose peptone 5.0 g / L, yeast powder 1.0 g / L, ferric citrate 0.1 g / L, sodium chloride 19.45 g / L, magnesium chloride 5.98 g / L, sodium sulfate 3.24 g / L, calcium chloride 1.8 g / L, potassium chloride 0.55 g / L, sodium carbonate 0.16 g / L, potassium bromide 0.08 g / L, strontium chloride 0.034 g / L, boric acid 0.022 g / L, sodium silicate 0.004 g / L, sodium fluoride 0.0024 g / L, sodium nitrate 0.0016 g / L, disodium hydrogen phosphate 0.008 g / L, and pH is 7.4-7.
8.
6. The preparation method of the ultraviolet absorber according to claim 2, wherein the bacterial fermentation culture medium formula is as follows: tryptone 10 g, yeast extract powder 5 g, sodium chloride 10 g, and sterile artificial seawater 1 L, wherein the artificial seawater formula is as follows: NaCl: 132.6 g, MgCl2·6H2O: 26.14 g, MgSO4: 16.53 g, CaCl2: 5.71 g, KCl: 3.63 g, NaHCO3: 1.01 g, NaBr: 0.42 g, and distilled water: 5000 mL, and pH is 7.0-7.
2.
7. An ultraviolet absorber characterized by comprising 3-Butyl-1-isobenzofuranone.
8. The use of the Kochia rhizogenes according to claim 1 in the preparation of anti-ultraviolet products.
9. The use of 3-butyl-1-isobenzofuranone as an ultraviolet absorber according to claim 7.
10. Use according to claim 9, wherein the compound is ###0002### 3-Butyl-1-isobenzofuranone absorbs ultraviolet rays with a wavelength of 200-400 nm.