Rhodobacter sphaeroides mutant strain with high yield of coenzyme q10 and application thereof

Abstract

The application discloses a mutant strain with high coenzyme Q10 yield, and has a preservation number of CGMCC No.22200.The mutant strain is obtained by enrichment, separation, mutagenesis and screening of Rhodobacter sphaeroides with high coenzyme Q10 content.The application also discloses application of the mutant strain in production of coenzyme Q10 and in medicines, health products, cosmetics and food.

Description

Technical Field

[0001] This invention belongs to the field of microbiology and relates to a high-yield coenzyme Q10-producing Rhodotorula globosum mutant strain and a method for producing coenzyme Q10. Background Technology

[0002] Coenzyme Q10, also known as ubiquinone, is a lipid-soluble quinone compound widely found on biological membranes. It is a yellow to orange-yellow crystalline powder, odorless and tasteless, and easily decomposes in light. Coenzyme Q10 is a natural antioxidant and cell metabolism activator produced by cells themselves. It is widely present in very low amounts in the cells of animals, plants, and microorganisms, and can be synthesized in all body tissues. Its biological activity mainly comes from the redox properties of the quinone ring and the physicochemical properties of the isoprene-like side chain. As an essential coenzyme for ATP production, coenzyme Q10 is responsible for proton transfer and electron transport in the respiratory chain, promotes oxidative phosphorylation and energy synthesis, and is an essential nutrient for maintaining normal metabolism and physiological functions. In addition, as a natural antioxidant, coenzyme Q10 can prevent the peroxidation of lipids, proteins, and DNA, maintaining the structural integrity and functional stability of biological membranes. Coenzyme Q10 is an essential element for human life, with a variety of physiological functions, such as: helping to provide sufficient oxygen to the myocardium and preventing sudden heart attacks; effectively preventing photoaging of the skin and reducing wrinkles around the eyes; used to prevent and treat chronic fatigue syndrome; promoting energy conversion and enhancing the function of the human immune system. Therefore, it is widely used in the pharmaceutical, health care, cosmetics and food industries.

[0003] The main methods for producing Coenzyme Q10 are chemical synthesis and microbial fermentation. Chemical synthesis involves complex procedures, produces a mixture of cis and trans isomers that are not easily absorbed by the human body, and has high purification costs. Microbial fermentation, on the other hand, offers advantages such as abundant raw material sources, low production costs, mild reaction conditions, natural products, good biological activity, and easy process control. It is a promising, efficient, economical method that is easily scaled up for large-scale industrial production. Furthermore, through strain selection and fermentation regulation techniques, the unit yield of the strain can be targeted to increase, and it has completely replaced chemical synthesis as the primary method for Coenzyme Q10 production.

[0004] In nature, strains that produce coenzyme Q10 mainly include those from the genera *Rhodospirillum*, *Agrobacterium*, *Paracococcus*, *Pseudomonas*, and *Candida*. *Rhodospirillum* bacteria of the family Rhodospirillumaceae obtain energy for growth and reproduction through cyclic photophosphorylation. They require a large accumulation of coenzyme Q10 within their cells to reduce free radicals generated during photosynthesis, making them one of the main strains used for large-scale fermentation production of coenzyme Q10. The biosynthetic pathway of coenzyme Q10 can be divided into three modules: the branched acid pathway—synthesis of the quinone ring nucleus; the side chain synthesis pathway (bacterial MEP pathway, fungal MVA pathway)—synthesis of the side chain; and the quinone ring modification pathway—condensation of two precursor substances and multi-step modification of the benzene ring. The biosynthetic pathway of coenzyme Q10 is long, involves many metabolic nodes, and has complex metabolic regulation. Natural strains that produce coenzyme Q10, usually screened from nature, typically have low fermentation levels and cannot meet the requirements for large-scale industrial fermentation production. Summary of the Invention

[0005] The purpose of this invention is to provide a high-yield coenzyme Q10-producing Rhodotorula globosum mutant strain and a method for producing coenzyme Q10 by fermentation.

[0006] This invention first isolated a strain of Rhodococcus pluvialis PSB8 from mud collected from lake water. Subsequently, based on strain PSB8, a mutant strain with high coenzyme Q10 production was successfully bred through mutagenesis breeding. The mutant strain was named Rhodococcus pluvialis TIB.RS20.

[0007] Therefore, in a first aspect, the present invention provides a high-yield coenzyme Q10-producing Rhodophyton floccosum strain (Rhodophyton spp.). Rhodobacter sphaeroides The mutant strain of ) was deposited on April 19, 2021, at the China General Microbiological Culture Collection Center (CGMCC, address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, postcode: 100101), with accession number CGMCC No. 22200.

[0008] Secondly, the present invention provides a method for producing coenzyme Q10, which involves fermenting the mutant strain, collecting the fermentation broth, and collecting the coenzyme Q10 therein.

[0009] More specifically, it includes the following steps:

[0010] (1) The mutant strain is cultured on a culture medium plate to obtain a single colony; preferably, the plate is activated again to obtain a single colony, so that the viability of the bacterial cells will be better.

[0011] (2) Scrape a single colony from the plate in step (1), inoculate it with seed culture medium, and shake to obtain seed liquid;

[0012] (3) Take the seed liquid from step (2) and transfer it to the fermentation medium and shake it for a certain period of time to obtain the fermentation broth; preferably, inoculate 3-7 mL of seed liquid with 50 mL of fermentation medium and shake it in a shake flask.

[0013] (4) Collect the fermentation broth from step (3) and detect coenzyme Q10 by liquid chromatography.

[0014] The culture temperature in step (1) is 25-35℃, preferably 32℃, and the culture time is 5-8 days, preferably 6 days.

[0015] Preferably, the single colony scraped in step (2) is a single colony on a 1 / 2 plate, the shaking speed during the shaking culture is 180-220 rpm, preferably 200 rpm, and the shaking culture time is 30-36 h, preferably 34 h;

[0016] Preferably, in step (3), the seed liquid is inoculated into every 50 ml of fermentation medium at a rate of 3-7 mL, preferably 5 mL, the shaking speed during culture is 180-220 rpm, preferably 200 rpm, and the shaking culture time is 90-120 h, preferably 110 h.

[0017] Thirdly, the present invention provides the application of the mutant strain in the production of coenzyme Q10.

[0018] Fourthly, the present invention provides the application of the mutant strain in pharmaceuticals, health products, cosmetics, and food.

[0019] The high-coenzyme Q10-producing mutant strain of *Rhodotorula globosum* of this invention was obtained through mutagenesis and screening of *Rhodotorula globosum* strains with high coenzyme Q10 content obtained through enrichment and isolation. This strain can produce high levels of coenzyme Q10. Experiments show that in shake-flask culture, the coenzyme Q10 yield of this strain reaches 182.4 mg / L, more than 10 times that of the original strain. It belongs to the category of high-coenzyme Q10-producing strains and has high application value. It can be used for coenzyme Q10 production and has a very broad prospect for application in medicine, health products, cosmetics, and food. Attached Figure Description

[0020] Figure 1 The standard curve of coenzyme Q10.

[0021] Figure 2 The yield of coenzyme Q10 from 11 isolated photosynthetic bacteria strains.

[0022] Figure 3 Microscopic image of strain PSB8.

[0023] Figure 4 Fatality curve.

[0024] Figure 5 The yield of coenzyme Q10 after 110 h of shake-flask fermentation of different green mutant strains.

[0025] Figure 6 Biomass of different green mutant strains after 110 h of shake-flask fermentation. Detailed Implementation

[0026] The following embodiments and accompanying drawings of the present invention are merely illustrative of specific implementation schemes for carrying out the present invention. These schemes and drawings should not be construed as limiting the present invention, and can be fully applied to various fields suitable for the present invention. For those skilled in the art, other modifications can be easily implemented. Therefore, any changes made without departing from the principles and essence of the present invention fall within the protection scope of the present invention.

[0027] Unless otherwise specified, the experimental techniques and methods used in this embodiment are conventional. Unless otherwise specified, all materials and reagents used in this embodiment can be obtained through legitimate commercial channels.

[0028] The present invention will now be described in further detail with reference to the accompanying drawings, so that those skilled in the art can implement it based on the description.

[0029] I. Culture medium formulation:

[0030] Enrichment and separation medium (g / L): yeast extract 10, dipotassium hydrogen phosphate 1.5, magnesium sulfate heptahydrate 0.3, ferrous sulfate heptahydrate 0.1, sodium chloride 2, manganese sulfate 0.02, choline chloride 0.0005, solid medium with agar powder 18, pH controlled at 7.0-7.4 by NaOH, and 10 mL of auxiliary solution added after sterilization.

[0031] Seed culture medium (g / L): yeast extract 1, anhydrous glucose 5, ammonium sulfate 1.5, corn steep liquor 1, potassium dihydrogen phosphate 0.3, dipotassium hydrogen phosphate 0.3, magnesium sulfate heptahydrate 1, ferrous sulfate heptahydrate 0.3, sodium chloride 2, manganese sulfate 0.001, choline chloride 0.001, calcium carbonate 5, pH controlled at 7.0-7.4 with NaOH, add 10mL of auxiliary solution after sterilization.

[0032] Fermentation medium (g / L): anhydrous glucose 40, ammonium sulfate 4, corn steep liquor 8, potassium dihydrogen phosphate 1.5, magnesium sulfate heptahydrate 5, ferrous sulfate heptahydrate 1, sodium chloride 2.5, manganese sulfate 0.02, choline chloride 0.002, calcium carbonate 10, solid medium with agar powder 18, pH controlled at 7.0-7.4 by NaOH, and 10 mL of auxiliary solution added after sterilization.

[0033] Excipients (g / L): Thiamine 0.5, Riboflavin 0.1, Pyridoxine 0.15, Niacin 1, Folic Acid 0.03, Biotin 0.04.

[0034] II. Detection Methods for Coenzyme Q10

[0035] The specific method is as follows:

[0036] (1) Sample pretreatment

[0037] After shaking the fermentation broth well, pipette 10 mL of the broth into a 100 mL brown volumetric flask, add 1-2 drops of 0.6 mol hydrochloric acid and 1 mL of hydrogen peroxide, shake gently, add 20 mL of acetone, and dilute to the mark with anhydrous ethanol. Sonicate at 35°C for 40 minutes. Centrifuge the entire sample at 5000 rpm for 15 minutes, remove the supernatant, evaporate to dryness using a rotary evaporator, dissolve in 2 mL of acetone, and filter through a 0.45 µm filter into a sample vial for liquid chromatography analysis. The amounts of hydrochloric acid, hydrogen peroxide, acetone, and anhydrous ethanol, as well as the volumetric flask volume, can be adjusted according to the volume of the fermentation broth.

[0038] (2) Preparation of standard products

[0039] Coenzyme Q10 standards of different concentrations (10 mg / L, 20 mg / L, 30 mg / L, 50 mg / L, 80 mg / L, 100 mg / L) were prepared using acetone as a solvent.

[0040] (3) HPLC detection conditions

[0041] The chromatographic column was an Agilent TC-C18, 4.6×250mm, 5u; the mobile phase was anhydrous ethanol:methanol = 1:1; the detection wavelength was 275nm; the column temperature was 35℃; the flow rate was 2.0mL / min; and the injection volume was 20µL.

[0042] (4) Construction of the Coenzyme Q10 standard curve

[0043] Standards of different concentrations were analyzed by HPLC under the conditions described above, and a standard curve of coenzyme Q10 concentration versus peak area was plotted. A standard curve for the HPLC detection of coenzyme Q10 concentration was plotted with coenzyme Q10 concentration (mg / L) on the x-axis and the measured peak area on the y-axis. Figure 1 As shown, the regression equation is y = 0.1678x + 0.0365, R0 2 =0.9985, the peak area showed a good linear relationship with the coenzyme Q10 concentration. After the sample liquid chromatography detection was completed, the coenzyme Q10 yield in the sample was calculated based on the coenzyme Q10 standard curve and the peak area of ​​the sample.

[0044] Example 1: Enrichment and Isolation of Photosynthetic Bacteria

[0045] Soil samples collected from lakeside water were anaerobically cultured under light using an anaerobic bottle containing 100 mL of enrichment and isolation medium. The lamp was a 40W incandescent bulb, 30-40 cm away, and the mixture was incubated at 32°C for 5-10 days, until the culture medium turned red to brownish-red. 1 mL of the culture was then transferred to another anaerobic bottle containing fresh enrichment and isolation medium and cultured again. This process was repeated 5-6 times to enrich photosynthetic bacteria.

[0046] The obtained bacterial suspension was serially diluted with liquid enrichment and separation medium. 100 μL of the bacterial suspension from each dilution was spread onto solid enrichment and separation medium and cultured in the dark aerobic environment at 32°C for 5-8 days until single colonies grew on the plates. Single colonies with bright color and short growth cycle were selected for repeated isolation and purification until a pure culture of photosynthetic bacteria was obtained.

[0047] Example 2: Determination of Coenzyme Q10 content in pure cultures

[0048] 1. Shake flask fermentation

[0049] Eleven pure-culture photosynthetic bacteria isolates were diluted appropriately and spread onto solid enrichment and isolation medium. They were then incubated aerobically in the dark at 32°C for 6 days to obtain single colonies. Each single colony was picked, diluted 100-fold with sterile water, and 100 μL was spread onto solid enrichment and isolation medium. The culture was then incubated aerobically in the dark at 32°C for 6 days to obtain fresh, activated single colonies.

[0050] Half of each single colony was scraped from the plate and inoculated into a shake flask containing 30 mL of seed culture medium. The culture was incubated at 32°C and 200 rpm for 34 h with shaking. 5 mL of the seed culture was then transferred to a shake flask containing 50 mL of fermentation culture medium. The culture was incubated at 32°C and 200 rpm for 110 h with shaking to obtain the fermentation broth. The coenzyme Q10 yield in the fermentation broth was determined using liquid chromatography. Each strain was performed in triplicate for shake flask fermentation.

[0051] 2. Determination of Coenzyme Q10 content

[0052] After processing, the samples were analyzed by liquid chromatography. The concentration of the samples was calculated based on the standard curve. The data from each group were compared, and the sample with the highest coenzyme Q10 content was selected. The results showed (see...) Figure 2 Of the 11 pure photosynthetic bacteria isolated (named PSB1, PSB2, PSB3, PSB4, PSB5, PSB6, PSB7, PSB8, PSB9, PSB10, and PSB11 respectively), PSB8 had the highest coenzyme Q10 content, at 16.6 mg / L.

[0053] Example 3: Observation of the morphological characteristics of the strain

[0054] 1. Observation of colony morphology characteristics

[0055] The strain PSB8, which had the highest coenzyme Q10 content obtained through enrichment and isolation, showed that after 6 days of dark aerobic culture on a solid enrichment and isolation medium, single colonies were red with a slightly lighter color at the periphery. The single colonies were round, smooth, slightly raised, with neat edges and a glossy appearance, and the diameter of the single colonies was between 1.5 and 2.5 mm. The suitable growth temperature was 25-35℃, and the suitable growth pH was 7.0-7.4.

[0056] 2. Observation of individual morphological characteristics of bacterial strains

[0057] The coenzyme Q10 enriched and isolated strain PSB8 was examined for its morphology using Gram staining and microscopic observation. Gram staining results indicated that PSB8 is a Gram-negative bacterium. Microscopic observation under both oil immersion eyepiece and objective lenses at 100x magnification revealed that most strains were spherical, with single cells ranging in size from 1.0µm to 1.3µm (see...). Figure 3 ).

[0058] Example 4: Identification of strain PSB8

[0059] 1. Sequencing of the 16S rRNA gene of the strain

[0060] The PSB8 strain was cultured and expanded according to the shake-flask fermentation method described in Example 2. Genomic DNA of the PSB8 strain was extracted according to the instructions on the bacterial genomic DNA extraction kit from Tiangen Biotech Co., Ltd. PCR amplification was performed using universal primers for the 16S rRNA gene synthesized by Tianjin Qingke Biotechnology Co., Ltd. (8F: 5'-AGAGTTTGATCCTGGCTCAG-3', 1492R: 5'-GGTTACCTTGTTACGACTT-3'). The PCR reaction system (50µL) included 25µL PrimeSTAR® Max DNA Polymerase, 1µL each of 8F and 1492R primers (10µM), 1µL PSB8 genomic DNA, and 22µL sterile ddH2O. PCR reaction conditions were: 98℃ for 2 min, 1 cycle; 98℃ for 10 s, 55℃ for 15 s, 72℃ for 30 s, 30 cycles; 72℃ for 5 min, 1 cycle; 4℃ forever. Sequencing was performed by Suzhou Genewiz Biotechnology Co., Ltd., using 8F and 1492R primers.

[0061] 2. 16S rRNA gene sequence analysis

[0062] The 16S rRNA gene fragment amplified by strain PSB8 was a single band, approximately 1.4 kb in size. Homology comparison of the 16S rRNA gene sequence of strain PSB8 with sequences in the GenBank database revealed that strain PSB8 naturally clustered with the 16S rRNA sequences of the genus *Rhodophyta*. It showed 99.85% similarity to three *Rhodophyta* strains with accession numbers EU649703.1, MG818312.1, and FM177579.1, indicating that strain PSB8 is most closely related to *Rhodophyta*.

[0063] Example 5: Determination of mutagenesis time by ambient pressure room temperature plasma (ARTP)

[0064] To obtain a relatively broad mutant library, the *Rhodopseudomonas spp.* strain PSB8 was subjected to ambient pressure room temperature plasma (ARTP) mutagenesis. First, the lethality of PBSB8 under plasma mutagenesis conditions was determined. PBSB8 cells seeded to mid-log phase were diluted with 0.8% NaCl solution to a concentration of 10⁻⁶ cells / mL. 8 A suspension of cells / mL was prepared, and 10 μL of the suspension was evenly spread onto an iron plate for ARTP mutagenesis. Mutagenesis was performed at 0 s, 20 s, 30 s, 40 s, 50 s, 60 s, 70 s, 80 s, and 90 s, with two replicates for each time point and three replicates for each time point plate application. After mutagenesis, the iron plate containing cells was placed in 1 mL of sterile water and vortexed to wash off the cells. The cell suspension was then serially diluted 10-fold to 10⁻⁶. -2 100 µL of diluted cell suspension was plated and incubated at 32°C for 6 days. Colony counts were then recorded. The lethality rate at different mutagenesis times was calculated using the following formula. A lethality rate curve was plotted with mutagenesis time on the x-axis and lethality rate at different mutagenesis times on the y-axis. The formula for calculating the lethality rate is: Lethality rate (%) = [(number of 0s mutagenesis colonies - number of Ns mutagenesis colonies) / number of 0s mutagenesis colonies] × 100%, where N = 20, 30, 40, 50, 60, 70, 80, 90.

[0065] from Figure 4 It can be seen that the lethality rate at 50s of mutagenesis is 74.86%, and the lethality rate at 80s of mutagenesis reaches more than 99.16%. When the lethality rate is 70%-80%, the probability of positive mutation after mutagenesis is the highest. Therefore, 50s was chosen as the mutagenesis time for the final construction of the mutant library.

[0066] Example 6: Construction of a mutant library

[0067] Take a concentration of 10 810 μL of a cell suspension (cells / mL) was spread onto iron plates for ARTP mutagenesis. The mutagenesis time was 50 s. Cells on two iron plates were mutagenized. After mutagenesis, the cells from both iron plates were resuspended in the same EP tube containing 1 mL of fermentation medium by vortexing. The vortexed cell suspension was then serially diluted 10-fold to 10⁻⁶. -2 100 μL of the diluted cell suspension was spread onto a solid fermentation medium plate and cultured aerobically in the dark at 32°C for 6 days. 420 single colonies grew, including 216 red colonies, 195 white colonies, and 9 green colonies.

[0068] Example 7: Screening of high-coenzyme Q10-producing mutants

[0069] 1. Shake flask fermentation conditions

[0070] To reduce the workload of screening, we first determined whether there were significant differences in the coenzyme Q10 production of colonies of different colors. Therefore, we picked 5 single colonies each of red, white and green on the plate, diluted them 100 times with sterile water, and then spread 100 μL of each on solid enrichment and separation medium. We then incubated them in the dark at 32°C for 6 days to obtain fresh and activated single colonies.

[0071] Half of each single colony was scraped from the plate and inoculated into a shake flask containing 30 mL of seed culture medium. The culture was incubated at 32°C and 200 rpm for 34 h with shaking. 5 mL of the seed culture was then transferred to a shake flask containing 50 mL of fermentation culture medium. The culture was incubated at 32°C and 200 rpm for 110 h with shaking to obtain the fermentation broth. The coenzyme Q10 yield in the fermentation broth was determined using liquid chromatography. Each strain was performed in triplicate for shake flask fermentation.

[0072] 2. Analysis of the coenzyme Q10 production capacity of mutant strains with different colony colors

[0073] After processing, the samples were analyzed by liquid chromatography. Samples with peak areas exceeding the standard curve range required dilution with acetone to an appropriate factor before further analysis. The concentration of the samples was calculated based on the standard curve, and the data from each group were compared. The results showed that green colonies consistently produced the highest levels of coenzyme Q10, followed by red colonies, while white colonies produced the lowest levels.

[0074] Example 8: Evaluation of shake-flask fermentation of different green mutant strains

[0075] 1. Shake-flask fermentation conditions for different green mutant strains

[0076] The original PSB8 strain, used as a control, was diluted appropriately and spread onto solid enrichment and isolation medium. It was then incubated aerobically in the dark at 32°C for 6 days to obtain single colonies. Each single colony was picked, diluted 100-fold with sterile water, and 100 μL was spread onto solid enrichment and isolation medium. The culture was then incubated aerobically in the dark at 32°C for 6 days to obtain fresh, activated single colonies.

[0077] Nine green single colonies (named LVM1-LVM9) were picked from the plate, diluted 100 times with sterile water, and then 100 μL of each colony were spread onto solid enrichment and separation medium and cultured in the dark at 32°C for 6 days to obtain fresh and activated single colonies.

[0078] Half of each single colony was scraped from the plate and inoculated into a shake flask containing 30 mL of seed culture medium. The culture was incubated at 32°C and 200 rpm for 34 h with shaking. 5 mL of the seed culture was then transferred to a shake flask containing 50 mL of fermentation culture medium. The culture was incubated at 32°C and 200 rpm for 110 h with shaking to obtain the fermentation broth. The coenzyme Q10 yield in the fermentation broth was determined using liquid chromatography. Each strain was performed in triplicate for shake flask fermentation.

[0079] 2. Comparison of Coenzyme Q10 production capacity and biomass of different green mutant strains

[0080] Compared with the control strain PSB8, the green mutant strain showed a significant increase in the ability to produce coenzyme Q10 (see Table 1 below). Figure 5 ).

[0081]

[0082] The LVM7 strain produced 78.6 mg / L of coenzyme Q10, a 4.6-fold increase compared to the control strain PSB8. Furthermore, the greener the single colony, the higher the coenzyme Q10 production. The mutant strain showed no significant difference in biomass compared to the control strain PSB8, indicating that the mutation did not affect bacterial growth (see [link to original text]). Figure 6 ).

[0083] Example 9: Mutagenesis of the green mutant strain LVM7 and screening of high-coenzyme Q10-producing strains

[0084] Based on the green mutant strain LVM7, Examples 6 and 8 were repeated three times, and finally a high-yielding coenzyme Q10 strain TIB.RS20 was obtained. The coenzyme Q10 yield in the shake flask reached 182.4 mg / L. The coenzyme Q10 yield of strain TIB.RS20 is 2.3 times that of LVM7 and 10.7 times that of PSB8. Therefore, it belongs to the high-yielding coenzyme Q10 strain and has high application value.

[0085] The strain TIB.RS20 was deposited on April 19, 2021, at the China General Microbiological Culture Collection Center (CGMCC, address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, postcode: 100101), with accession number CGMCC No. 22200. sequence list <110> Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences <120> A high-coenzyme Q10-producing mutant strain of Rhodopseudomonas aeruginosa and its application <160> 2 <170> SIPOSequenceListing 1.0 <210> 1 <211> 20 <212> DNA <213> Artificial sequence() <400> 1 agagtttgat cctggctcag 20 <210> 2 <211> 20 <212> DNA <213> Artificial sequence() <400> 2 ggttaccttg ttacgactt 19

Claims

1. A mutant strain that produces high levels of coenzyme Q10, characterized in that, The mutant strain is Rhodobacter phaeroides, which was deposited at the China General Microbiological Culture Collection Center (CGMCC) on April 19, 2021, with accession number CGMCC No. 22200.

2. A method for preparing coenzyme Q10, characterized in that, The mutant strain of claim 1 is fermented, the fermentation broth is collected, and the coenzyme Q10 therein is collected.

3. The method as described in claim 2, characterized in that, The fermentation culture temperature is 25-35℃.

4. The method as described in claim 2 or 3, characterized in that, The fermentation culture is a shaking culture with a rotation speed of 180-220 rpm and a shaking culture time of 90-120 h.

5. The method as described in claim 2, characterized in that, The steps before fermentation are as follows: (1) The mutant strain of claim 1 is cultured on a culture medium plate to obtain a single colony; (2) Scrape a single colony from the plate in step (1), inoculate it with seed culture medium, and shake to obtain seed liquid; (3) Take the seed liquid from step (2) and transfer it to the fermentation medium.

6. The method as described in claim 5, characterized in that, After obtaining a single colony in step (1), it is cultured again on a culture medium plate to obtain a single colony.

7. The method as described in claim 5, characterized in that, In step (3), the seed culture is inoculated at a ratio of 3-7 mL of seed culture to 50 mL of fermentation medium, and then cultured in a shake flask with shaking.

8. The application of the mutant strain as described in claim 1 in the production of coenzyme Q10.

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