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WO2008063020A9 - Souche de rhodobacter sphaeroides sk2h2 présentant une teneur élévée en coenzyme q10 et procédé de production de coenzyme q10 à l'aide de ladite souche - Google Patents

Souche de rhodobacter sphaeroides sk2h2 présentant une teneur élévée en coenzyme q10 et procédé de production de coenzyme q10 à l'aide de ladite souche

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Publication number
WO2008063020A9
WO2008063020A9 PCT/KR2007/005904 KR2007005904W WO2008063020A9 WO 2008063020 A9 WO2008063020 A9 WO 2008063020A9 KR 2007005904 W KR2007005904 W KR 2007005904W WO 2008063020 A9 WO2008063020 A9 WO 2008063020A9
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Prior art keywords
strain
coenzyme qlo
sphaeroides
coenzyme
sk2h2
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Ceased
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PCT/KR2007/005904
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English (en)
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WO2008063020A1 (fr
Inventor
Nahm Ryune Cho
Min Soo Park
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SK Energy Co Ltd
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SK Energy Co Ltd
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Publication of WO2008063020A9 publication Critical patent/WO2008063020A9/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/66—Preparation of oxygen-containing organic compounds containing the quinoid structure
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales

Definitions

  • the present invention relates to a Rhodobacter sphaeroides strain having a high content of coenzyme QlO and a method of producing coenzyme QlO using the same, and more particularly, to a novel Rhodobacter sphaeroides (hereinafter, referred to as R. sphaeroides) SK2H2 strain having a highly increased content of coenzyme QlO, the strain being obtained by mutating a R. sphaeroides strain through genetic modification using mutagens and selecting its mutant strains having an increased activity, and a method of producing coenzyme QlO whose productivity is improved when the R. sphaeroides strain is cultured under a certain culture condition.
  • R. sphaeroides novel Rhodobacter sphaeroides
  • CoQlO Coenzyme QlO
  • Ubiquinone 10 Coenzyme QlO
  • 2,3-dimethoxy-dimethyl-6-decaprenyl-l,4-benzoquinone which refers to fat-soluble benzoquinone having an isoprenoid side chain composed of C- 5 isoprenoid unit as represented by the following Formula 1.
  • Coenzyme QlO is widely present in microorganisms, plants and animals (including human) and takes an important role in maintaining a healthy state of living organisms and protecting them from a variety of diseases.
  • the coenzyme QlO is an essential coenzyme in the electron transport system of mitochondria which takes an important role in ATP biosynthesis and physiologically functions to prevent the oxidation of DNA, lipids, proteins, and the like in the cells as a fat soluble antioxidant.
  • the coenzyme QlO has been proven to be a compound that is effective to prevent and treat heart diseases, degenerative nerve disorders, etc, it is expected that the coenzyme QlO will be increasingly used for medicines or health food supplements (Zahiri, H. S. et al., Metabolic engineering, 8, 406-416, 2006).
  • the method of producing above-mentioned coenzyme QlO includes 1) an extraction from natural substances such as animal/plant tissues, 2) a chemical synthesis, and 3) a fermentation synthesis by microorganisms.
  • the fermentation synthesis includes 1) a method using microorganisms that spontaneously produce coenzyme QlO, 2) a method using microorganisms that are endowed with an ability to synthesize coenzyme QlO through the genetic modifications, and 3) a method including: inducing mutation in microorganisms that spontaneously produce coenzyme QlO, screening strains having an improved activity and optimizing culture conditions of the selected strains, all of which have been highly studied in the art.
  • Cheong et al. reported a method where coenzyme QlO is produced in a yield of 642 mg/L by transforming a strain Agrobacterium tumefaciens with 1-deoxy-D-xylulose 5-phosphate synthase and decaprenyl diphosphate synthase gene and optimizing a fermentation process of the transformed strain, wherein a content of the coenzyme QlO is 7.3 mg/gDCW (Cheong, S. R. et al., US 2005/0181490 (2005)).
  • the coenzyme QlO content per dry cell weight was increased up to 139 mg/gDCW, which is significantly improved compared to that of the wild-type strain, but it is not easy to develop effective processes due to the low content of coenzyme QlO produced in the transformant strains.
  • mutant strains having increased activity were screened using compounds, such as paraquat or linolenic acid, that suppress respiratory chain activity of the electron transport system, the selected mutant strain had a yield that is higher about 3.7 times than the wild-type strain, but a content of coenzyme QlO was 3.5 mg/gDCW and a yield of the mutant strain was 229 mg/L (Pfaller R. et al., US2004 /0209368 (2004)).
  • the present inventors have ardently attempted to solve the problems regarding the low content of coenzyme QlO in the fermentation process as described above, and therefore they found that a content of coenzyme QlO in a mutant strain of a Rhodobacter sp. strain, particularly a R. sphaeroides KCCM35488 strain may be highly increased by inactivating a crtE gene of the R. sphaeroides KCCM35488 strain, mutating genes of the R. sphaeroides KCCM35488 strain with a chemical mutagen NTG (N-methyl- N'-nitro-N-nitrosoguanidine), selecting a strain R. sphaeroides SK2H2 having improved activities, and modifying its culture methods, and therefore the present invention was completed on the basis of the above facts. [43]
  • An aspect of the present invention provides a novel Rhodobacter sphaeroides strain having a high content of coenzyme QlO. [45] [46] Another aspect of the present invention provides a method of producing coenzyme
  • Rhodobacter sphaeroides SK2H2 (Accession No.: KCTC10915BP) strain characterized in that it has a high content of coenzyme QlO.
  • a method of producing coenzyme QlO characterized in that the method includes: culturing the
  • Rhodobacter sphaeroides SK2H2 (Accession No.: KCTC10915BP) and isolating coenzyme QlO from a culture broth of the strain.
  • a strain R. sphaeroides SK2H2 having a high intracellular content of coenzyme QlO may be obtained by inactivating a crtE gene of the R. sphaeroides KCCM35488 strain, treating the crtE gene-inactivated strain with a chemical mutagen NTG (N-methyl- N'-nitro-N-nitrosoguanidine) to produce its randomly mutated strains and selecting from the randomly mutated strains a R. sphaeroides strain having a high content of coenzyme QlO per dry cell weight.
  • the coenzyme QlO may be effectively produced by culturing the selected strain prepared thus, and isolating coenzyme QlO from its culture broth.
  • the method of producing a R. sphaeroides SK2H2 having a high content of coenzyme QlO according to the present invention proceeds by removing some of a crtE gene of a R. sphaeroides KCCM35488 strain by using a homologous recombination method, and will be described in detail, as follows.
  • the crtE is a gene that produces geranylgeranyl pyrophosphate synthase as a reaction product, and removal of some or the whole of the gene inhibits the synthesis of carotenoid.
  • a partial DNA fragment of the crtE gene is amplified with a pair of primers with bases complementary to the crtE gene, and the amplified DNA fragment is then isolated and purified.
  • the purified DNA fragment is cut with specific restriction enzymes, and inserted into a vector plasrrid that has been cut with the same restriction enzymes to construct a vector for homologous recombination, and E. coli is transformed with the vector to prepare a transformed E. coli strain.
  • the plasrrid vector in the transformed E. coli is transferred into the R. sphaeroides KCCM35488 strain through the conjugation of the transformed E. coli and the R. sphaeroides KCCM35488.
  • the strains prepared by introducing the vector onto chromosomes of the R. sphaeroides KCCM35488 through the homologous recombination, may be selected in the form of colonies since they survive in continuous cultures in an LB agar medium supplemented with 20 ⁇ g/ml of antibiotic Kanamycin and an LB agar medium supplemented with 7% sucrose.
  • the selected colonies are used as templates, and crtE gene-inactivated mutant strains are screened and confirmed by the polymerase chain reaction using a set of primers, and the resulting crtE gene-inactivated mutant strain is represented by R. sphaeroides KCCM35488 ( ⁇ c rtE).
  • the crtE gene-inactivated R. sphaeroides KCCM35488 ( ⁇ c rtE) strain is cultured in a nutrient medium and collected when an optical density (hereinafter, referred to as O.D., at a wavelength of 600 nm) of the strain is in a range from 0.7 to 1.0 when O.D. is measured at a wavelength of 600 nm using a spectrophotometer, and a mutagen- containing butter is added to the collected cells, and then cultured at 3O 0 C for 10 minutes.
  • O.D. optical density
  • mutant cells are collected, washed several times with buffer to remove the mutagen off, diluted at a suitable ratio with buffer, and then plated on a medium with selection pressure to screen viable strains.
  • the viable strains are cultured in a nutrient medium again, and collected.
  • coenzyme QlO is extracted with a solvent and its content is quantitatively analyzed with HPLC to screen a strain having the highest content of coenzyme QlO.
  • the selected strain is named Rhodobacter sphaeroides (R. sphaeroides) SK2H2, and deposited in the Korean Collection for Type Cultures (KCTC), and its Accession number is KCTC10915BP (deposited on March 3, 2006).
  • the mutagen, used in the manufacture of the strain according to the present invention includes N-methyl-N'-nitro-N-nitroso-guanidine (NTG), ethyl nitroso-urea (ENU), ethylmethane sulfonate (EMS), dimethyl sulfate (DMS), ethidium bromide (EtBr), acridine orange, ultraviolet (UV), etc., and NTG is widely used among them.
  • NTG N-methyl-N'-nitro-N-nitroso-guanidine
  • ENU ethyl nitroso-urea
  • EMS ethylmethane sulfonate
  • DMS dimethyl sulfate
  • EtBr ethidium bromide
  • UV ultraviolet
  • the medium used to screen mutant strains includes a medium including compounds that can give a selection pressure to the nutrient medium.
  • the compound used herein include intermediates in the biosynthesis of coenzyme QlO, structural analogues of the coenzyme QlO, respiratory inhibitors that inhibit the electron transport system, oxidizing agents that induce the increase in antioxidant activity, etc.
  • the intermediates of the coenzyme QlO include 4-hydroxy benzoic acid, chorisnic acid, shiMnic acid, etc., and 1,4-benzoquinone, 2,6-dimethyl benzoquinone, 2,3-dimethoxy-5-methyl benzoquinone, and the like function as the structural analogues of the coenzyme QlO.
  • Menadione (2-methyl-l,4-naphthoquinone), antimycin A, and the like are used as the respiratory inhibitor, and paraquat (l,l'-dimethyl-4,4'-bipyridinium), linolenic acid, or the like is used as the oxidizing agent that functions to increase the intracellular oxidation reaction.
  • the Rhodobacter sphaeroides SK2H2 strain selected through the above-mentioned method is suitably cultured in a nutrient medium supplemented with molasses and glycerol, a content of the coenzyme QlO per dry cell weight is increased to greater than 20 mg/gDCW.
  • the Rhodobacter sphaeroides SK2H2 strain may be cultured in a medium containing bacto tryptone 1.6%, yeast extract 1%, sodium chloride 0.5%, molasses 4%, and glycerol 2%.
  • the coenzyme QlO may be easily isolated and quantified according to the conventional methods.
  • the cultured cells are collected from the culture medium through cen- trifugation, compounds such as CellyticB solution (Sigma), that lyses cells, and lysozme are added to the collected cells, nixed thoroughly, and cultured at 37 0 C.
  • a solvent in which hexane and propanol are nixed in an appropriate ratio is added to the culture medium.
  • the resulting culture medium is stirred, and then centrifuged to collect a supernatant from two separated layers.
  • the collected supernatant is dried using a dryer to remove solvents off, and then dissolved in ethanol completely, and the resulting mixture is analyzed with high-performance liquid chromatography (HPLC).
  • HPLC high-performance liquid chromatography
  • Mghtysil RP- 18 GP 150-4.6 (5/M) of Kanto Chemical (Japan) is used as a column used in the liquid chromatographic analysis, and a mixture solution of methanol and ethanol at a ratio 1:1 (vol.: vol.) is used as an eluant.
  • the cultured product is detected at a wavelength of UV 275 nm while the mixture solution flows through the column at a rate of 1 mi per minute.
  • the cultured product, coenzyme QlO may be easily distinguished since the coenzyme QlO is detected at a time point of 16.7 minutes.
  • the coenzyme QlO may be quantified by calculating the measured values using an equation obtained through a standard curve.
  • the content of the coenzyme QlO per dry cell content is 20 greater than mg/gDCW, which is increased 40% or more, compared to the highest content of the coenzyme QlO in the conventional strains and its process, and therefore the strain used in the present invention may be used to develop an effective commercial process since it is possible to produce a high concentration of the coenzyme QlO when the strain is cultured in a large scale through the fed-batch culture.
  • a pH range from 6.5 to 8.5 is suitable in the culture of the strain.
  • pH is less than 6.5, or greater than 8.5, the content of the coenzyme QlO per dry cell weight may be reduced.
  • its culture temperature is preferably in a range from 25 to 4O 0 C. A culture rate of the strain is so slow when the culture temperature is less than 25 0 C, whereas a content of the coenzyme QlO per dry cell weight is reduced when the culture temperature exceeds 4O 0 C.
  • volume of the culture medium should be also used at a suitable volume range.
  • Volume of the medium in the culture of the strain is preferably in a range from 20 to 40 ml on the basis of a 25OmL Erlenmeyer flask, but the present invention is not particularly United thereto.
  • the culture time is preferably in a range from 72 to 96 hours.
  • a content of the coenzyme QlO per dry cell weight is in a range of to greater than 20 mg/gDCW when the R. sphaeroides SK2H2 strain is cultured under the above-mentioned conditions.
  • Example 1 Inactivation of crtE gene of R. sphaeroides KCCM35488 strain
  • the R. sphaeroides KCCM35488 strain kindly provided by Korean culture center of microorganisms (KCCM), was used in this experiment, and a crtE gene of the R. sphaeroides KCCM35488 strain was inactivated by removing some of the crtE gene on its chromosomes using a homologous recombination method in this experiment.
  • primers were synthesized, as follows.
  • crtE-n (5'-CATGCTGCAGCGTTGAAGACGATGTGATCG)
  • crtE-rl (5'-TTCAGCCGCTCATCTTGTGCCTCATAGGACA)
  • crtE-f2 (5'-GCACAAGATGAGCGGCTGAAGGACATCCTC)
  • crtE-xl ACTGCTGCAGTGTCAACTTTATTGGACAGT
  • the crtE-fl and crtE-rl primers were used to amplify a DNA fragment of about
  • PCR polymerase chain reaction
  • the amplified DNA fragment was isolated and purified using a Qiagen PCR purification Mt (Qiagen).
  • the crtE- ⁇ 2 and crtE-r2 primers were used to amplify a DNA fragment of about 1.4 kb from the R. sphaeroides KCCM35488 through a polymerase chain reaction (PCR).
  • the amplified DNA fragment was isolated and purified using a Qiagen PCR purification Mt (Qiagen).
  • the two DNA fragments with about 1.0 kb and 1.4 kb obtained through the polymerase chain reaction were put into one nicrotube and nixed, and the crtE-fl and crtE-r2 primers were added to the resulting DNA fragment mixture, and the DNA fragment mixture was then subject to the polymerase chain reaction to amplify a DNA fragment of about 2.4 kb.
  • the amplified DNA fragment was isolated and purified using a Qiagen PCR purification Mt, and then cut with a restriction enzyme PstI.
  • the DNA fragment of about 2.4 kb cut by the PstI was introduced into a pLOl plasnid (Kmr sacB, RP4 oriT, CoIEl ori) (Lenz, O. et al., J. Bacteriol., 176:4385-4393, 1994) that has been cut by the same restriction enzyme, and the resulting plasnid was named pLOl-dcrtE, and thus the manufacture of a vector for homologous recombination was completed.
  • E. coli S 17- 1 Tra+ recA pro thi hsdR chr::RP4-2) (Simon, R.
  • sphaeroides KCCM35488 through a single crossover event were screened on an LB agar medium (1.0 % bacto tryptone, 0.5 % yeast extract, 1.0 % sodium chloride, 1.5% agar) supplemented with 20 ⁇ g/ml of antibiotic Kanamycin.
  • the selected colonies were put into an LB medium (1.0 % bacto tryptone, 0.5 % yeast extract, 1.0 % sodium chloride) and suspended, and the resulting suspension was plated on an LB agar medium supplemented with 7% sucrose.
  • Colonies grown on the sucrose-containing LB agar medium are colonies that have genetic characters where the pLOl-dcrtE plasnid introduced into the chromosomes is left out from the chromosomes through a double crossover event.
  • these colonies there are two types of genetic characters; one wild type in which the crtE gene remains intact, and the other mutant type in which a sequence of about 0.7 kb is removed from the crtE gene.
  • the crtE gene was divided into a mutant type when a DNA fragment of about 2.4 kb was detected, and divided into a wild type when a DNA fragment of about 3.1 kb was detected.
  • the colonies having the DNA fragment of 2.4 kb were finally confirmed through the polymerase chain reaction, and selected as the c/tE-inactivated strain of the R. sphaeroides KCCM35488, and then represented by R. sphaeroides KCCM35488 ( ⁇ c rtE).
  • optical density (O.D.) of the resulting LB broth was in a range from 0.7 to 1.0 when the optical density (O.D.) was measured with a spectrophotometer at 600 nm
  • 1 ml of the LB broth was put into a nicrotube, cen trifuged at a rotary speed of 13,000 rpm for 5 minutes to collect the cells, and the cells were washed with the same equivalent amount of phosphate buffer (0.8% sodium chloride, 0.02% potassium chloride, 0.144 % sodium phosphate, dibasic, 0.024%, potassium dihydrogen phosphate, pH 7.4), and then recovered.
  • phosphate buffer 0.8% sodium chloride, 0.02% potassium chloride, 0.144 % sodium phosphate, dibasic, 0.024%, potassium dihydrogen phosphate, pH 7.4
  • a phosphate buffer containing NTG as mutagen at a concentration range from 0.001 to 0.01 mg/ml was added to the recovered cells, and the cells were incubated at 3O 0 C for 10 minutes.
  • the resulting cell broth was centrifuged at a rotary speed of 13,000 rpm for 5 minutes to collect the cells, and the collected cells were washed with 1 ml of phosphate buffer twice to remove the mutagen off, and then re- suspended in 1 ml of phosphate buffer.
  • the resulting suspension was sequentially diluted 10-fold, and each of the diluted solutions was plated on an LB agar medium supplemented with 4-hydroxy benzoic acid as selection pressure for screening strains with an increased biosynthetic activity of coenzyme QlO, and then incubated for 4 days to obtain about 200 mutant strains.
  • the culture broth was inoculated at a concentration of 2% into a 250 ml Erlenmeyer flask containing 30 ml of coenzyme QlO culture medium (1.6 % bacto tryptone, 1.0 % yeast extract, 0.5 % sodium chloride, 4% molasses, 2% glycerol, pH 7.0), and then incubated at 3O 0 C for 3 days in a shaking incubator (200rpm).
  • coenzyme QlO culture medium 1.6 % bacto tryptone, 1.0 % yeast extract, 0.5 % sodium chloride, 4% molasses, 2% glycerol, pH 7.0
  • Dry cell weight (g/L) Weight of dish before dryness (mg) - Weight of dish after dryness (mg) / 5
  • a method of measuring a yield (mg/L) of coenzyme QlO is described, as follows. After the cell culture, 0.5 ml of the cell culture broth was put into a nicrotube and centrifuged at a rotary speed of 13,000 rpm for 5 minutes to collect the cells. 1 ml of phosphate buffer was added to the collected cells, washed thoroughly, and then centrifuged to collect the cells. 1 ml of 20 mM Tris-HCl buffer (pH 7.6) was added to the nicrotube containing the collect cells, suspended homogeneously, and then centrifuged again to collect the cells.
  • Analytic conditions of the liquid chromatography are as follows.
  • the liquid chro- matography used herein was HP 1050 Series Liquid Chromatography System (Hewlett Packard), Mghtysil RP- 18 GP 150-4.6 (5/M) from Kanto Chemical (Japan) was used as a column, and a mixture solution of methanol and ethanol at a ratio 1:1 (vol.: vol.) was used as an eluant, and flow rate of the eluant was 1 ml per minute.
  • the cultured product was detected at a wavelength of UV 275 nm.
  • the cultured product, coenzyme QlO may be easily distinguished since the coenzyme QlO was detected at a time point of 16.7 minutes.
  • the coenzyme QlO was quantified by calculating the measured values using an equation obtained through a standard curve.
  • the reference coenzyme QlO Aldrich
  • the reference coenzyme QlO Aldrich
  • the content (mg/gDCW) of the coenzyme QlO was calculated according to the following Equation 3 by using the dry cell weight (gDCW/L) and yield (mg/L) of the coenzyme QlO.
  • Each of the 200 R. sphaeroides SK2H2 strains was measured under the above- mentioned conditions for dry cell weight (gDCW/L), yield (mg/L) and content (mg/gDCW), and the results from the R. sphaeroides KCCM35488 strain and the R. sphaeroides KCCM35488 ( ⁇ c rtE) strain were then compared to each other to select 8 strains with a 10% increased activity. The results are listed in the following Table 1.
  • the R. sphaeroides SK2H2 strain having the highest content of coenzyme QlO was selected, and deposited in the Korean Collection for Type Cultures (KCTC), and its accession number was KCTC10915BP (deposited on March 3, 2006).
  • Example 4 Dry cell weight, Yield and Content of R. sphaeroides SK2H2 according to the Culture time
  • a colony of the R. sphaeroides SK2H2 strain prepared in Example 3 was inoculated in a test tube containing 3 ml of LB medium, and cultivated at 3O 0 C for 24 hours in a shaking incubator (200 rpm).
  • the culture broth was inoculated at a concentration of 2% into a 250 ml Erlenmeyer flask containing 30 ml of coenzyme QlO culture medium (1.6 % bacto tryptone, 1.0 % yeast extract, 0.5 % sodium chloride, 4% molasses, 2% glycerol, pH 7.0), and then cultivated at 30°Cfor 4 days in a shaking incubator (200rpm).
  • the cell culture broth was taken, and its dry cell weight (gDCW/L), yield (mg/L) and content (mg/gDCW) were calculated in the same manner as in Example 3. The results are listed in the following Table 2.
  • the Rhodobacter sphaeroides SK2H2 strain (Accession No.: KCTC 10915BP) according to the present invention has an extremely high content of coenzyme QlO per dry cell weight, which is greater than 20 mg/gDCW, compared to the previously produced strains, and therefore it is possible to produce coenzyme QlO effectively in a large-scaled culture of the strain.
  • the microorganism identified under 1 above was accompanied by.
  • microorganism identified under I above was received by this International Depositary Authority on and a request to convert the original deposit to a deposit under the Budapest Treaty was received by it on

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Abstract

La présente invention concerne une souche de Rhodobacter sphaeroides SK2H2 (numéro d'enregistrement : KCTC10915BP) présentant une teneur élevée en coenzyme Q10, ainsi qu'un procédé de production de coenzyme Q10 au moyen de ladite souche. Le procédé consiste à cultiver la souche de Rhodobacter sphaeroides SK2H2 (numéro d'enregistrement : KCTC10915BP) dotée d'une teneur élevée en coenzyme Q10, et à isoler la coenzyme Q10 d'un bouillon de culture de cette souche. La souche de Rhodobacter sphaeroides SK2H2 (numéro d'enregistrement : KCTC10915BP) présente une teneur très élevée de coenzyme Q10 en poids de matières cellulaires sèches par comparaison avec les souches produites auparavant, ladite teneur étant supérieure à 20 mg/gDCW, et il est donc possible de produire efficacement la coenzyme Q10 dans une culture de la souche à grande échelle.
PCT/KR2007/005904 2006-11-24 2007-11-22 Souche de rhodobacter sphaeroides sk2h2 présentant une teneur élévée en coenzyme q10 et procédé de production de coenzyme q10 à l'aide de ladite souche Ceased WO2008063020A1 (fr)

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