Processes for producing coenzyme q10

Abstract

The present invention relates to a process for producing reduced coenzyme Q 10 which comprises obtaining microbial cells containing reduced coenzyme Q 10 at a ratio of not less than 70 mole % among the entire coenzymes Q 10 , optionally disrupting the cells and recovering thus-produced reduced coenzyme Q 10 . The present invention also relates to a process for producing oxidized coenzyme Q 10 which comprises either recovering oxidized coenzyme Q 10 after oxidizing the above-mentioned microbial cells or disrupted product thereof, or recovering reduced coenzyme Q 10 from the above-mentioned microbial cells or disrupted product thereof to oxidize thus-obtained reduced coenzyme Q 10 thereafter. According to the processes of the present invention, reduced coenzyme Q 10 and oxidized coenzyme Q 10 can be produced simply on the industrial scale.

1 . A process for producing the reduced coenzyme Q 10  represented by the following formula (I):  
       
         
           
           
               
               
           
         
         which comprises culturing reduced coenzyme Q 10 -producing microorganisms in a culture medium containing a carbon source, a nitrogen source, a phosphorus source and a micronutrient to obtain microbial cells containing reduced coenzyme Q 10  at a ratio of not less than 70 mole % among the entire coenzymes Q 10 ,  
         optionally disrupting the microbial cells and  
         extracting thus-produced reduced coenzyme Q 10  by an organic solvent:  
       
     
     
         2 . The process according to  claim 1 , 
 wherein the reduced coenzyme Q 10  is contained at a ratio of not less than 70 mole % among the entire coenzymes Q 10 .    
     
     
         3 . (Canceled)  
     
     
         4 . The process according to  claim 1 , 
 wherein the culture is carried out at 15 to 45° C. and at a pH of 4 to 9.    
     
     
         5 . The process according to  claim 1 , 
 wherein the concentration of the carbon source in the culture is controlled to a concentration that no adverse effects are substantially caused on the productivity of reduced coenzyme Q 10 .    
     
     
         6 . (Canceled)  
     
     
         7 . (Canceled)  
     
     
         8 . (Canceled)  
     
     
         9 . (Canceled)  
     
     
         10 . (Canceled)  
     
     
         11 . (Canceled)  
     
     
         12 . (Canceled)  
     
     
         13 . (Canceled)  
     
     
         14 . (Canceled)  
     
     
         15 . (Canceled)  
     
     
         16 . The process according to  claim 1 , 
 wherein the extraction of reduced coenzymes Q 10  is carried out from wet cells or dry cells of the microbial cells or disrupted product thereof by using a hydrophilic organic solvent.    
     
     
         17 . (Canceled)  
     
     
         18 . The process according to  claim 1 , 
 wherein the extraction of the reduced coenzymes Q 10  is carried out from an aqueous suspension of the microbial cells or disrupted product thereof by using a hydrophobic organic solvent.    
     
     
         19 . The process according to  claim 18 , 
 wherein the hydrophobic organic solvent is a hydrocarbon, a fatty acid ester or an ether.    
     
     
         20 . The process according to  claim 18 , 
 wherein the hydrophilic organic solvent is used as an auxiliary solvent in combination with the hydrophobic organic solvent.    
     
     
         21 . The process according to  claim 20 , 
 wherein the hydrophobic organic solvent is a hydrocarbon, and the hydrophilic organic solvent is an alcohol.    
     
     
         22 . (Canceled)  
     
     
         23 . (Canceled)  
     
     
         24 . The process according to  claim 20 , 
 wherein the extraction is carried out under the condition that the hydrophobic organic solvent is contained in 25 to 65% by volume and the hydrophilic organic solvent is contained in 5 to 50% by volume.    
     
     
         25 . (Canceled)  
     
     
         26 . (Canceled)  
     
     
         27 . (Canceled)  
     
     
         28 . (Canceled)  
     
     
         29 . The process according to  claim 1 , 
 wherein the reduced coenzyme Q 10  is contained at a ratio of not less than 70 mole % among the entire coenzymes Q 10      in the case that the reduced coenzyme Q 10 -producing microorganisms are cultured with shaking (amplitude: 2 cm, 310 reciprocation/min) at 25° C. for 72 hours in 10 mL of a culture medium [(glucose: 20 g, peptone: 5 g, yeast extract: 3 g, malt extract: 3 g)/L, pH: 6.0] using a test tube (inner diameter: 21 mm, entire length: 200 mm),    the obtained broth is optionally concentrated,    the obtained solution is vigorously shaken for 3 minutes using 10 parts by volume of glass beads (425 to 600 μm) to disrupt the microorganisms under a nitrogen atmosphere in the concomitant presence of 3 parts by volume of isopropanol and 18.5 parts by volume of n-hexane relative to 10 parts by volume of the broth, and    the prepared hydrophobic organic solvent phase (n-hexane phase) is analyzed by HPLC.    
     
     
         30 . The process according to  claim 29 , 
 wherein the reduced coenzyme Q 10 -producing microorganisms have not less than 1 μg/mL of a productivity of reduced coenzyme Q 10  per unit culture medium when measured by HPLC under the condition according to  claim 29 .    
     
     
         31 . The process according to  claim 30 , 
 wherein the microorganisms are microorganisms of the genus  Agrobacterium,  the genus  Aspergillus,  the genus  Acetobacter,  the genus  Aminobacter,  the genus  Agromonas,  the genus  Acidiphilium,  the genus  Bulleromyces,  the genus  Bullera,  the genus  Brevundimonas,  the genus  Cryptococcus,  the genus  Chionosphaera,  the genus  Candida,  the genus  Cerinosterus,  the genus  Exisophiala,  the genus  Exobasidium,  the genus  Fellomyces,  the genus  Filobasidiella,  the genus  Filobasidium,  the genus  Geotrichum,  the genus  Graphiola,  the genus  Gluconobacter,  the genus  Kockovaella,  the genus  Kurtzmanomyces,  the genus  Lalaria,  the genus  Leucosporidium,  the genus  Legionella,  the genus  Methylobacterium,  the genus  Mycoplana,  the genus  Oosporidium,  the genus  Pseudomonas,  the genus  Psedozyma,  the genus  Paracoccus,  the genus  petromyces,  the genus  Rhodotorula,  the genus  Rhodosporidium,  the genus  Rhizomonas,  the genus  Rhodobium,  the genus  Rhodoplanes,  the genus  Rhodopseudomonas,  the genus  Rhodobacter,  the genus  Sporobolomyces,  the genus  Sporidiobolus,  the genus  Saitoella,  the genus  Schizosaccharomyces,  the genus  Sphingomonas,  the genus  Sporotrichum,  the genus  Sympodiomycopsis,  the genus  Sterigmatosporidium,  the genus  Tapharina,  the genus  Tremella,  the genus  Trichosporon,  the genus  Tilletiaria,  the genus  Tilletia,  the genus  Tolyposporium,  the genus  Tilletiopsis,  the genus  Ustilago,  the genus  Udeniomyces,  the genus  Xanthophilomyces,  the genus  Xanthobacter,  the genus  Paecilomyces,  the genus  Acremonium,  the genus  Hyhomonus,  or the genus  Rhizobium.      
     
     
         32 . (Canceled)  
     
     
         33 . A process for producing the oxidized coenzyme Q 10  represented by the following formula (II):  
       
         
           
           
               
               
           
         
         which comprises culturing reduced coenzyme Q 10 -producing microorganisms in a culture medium containing a carbon source, a nitrogen source, a phosphorus source and a micronutrient to obtain microbial cells containing reduced coenzyme Q 10  at a ratio of not less than 70 mole % among the entire coenzymes Q 10 ,  
         optionally disrupting the microbial cells; and  
         either oxidizing thus-produced reduced coenzyme Q 10  to oxidized coenzyme Q 10  and then extracting the resultant by an organic solvent, or extracting thus-produced reduced coenzyme Q 10  by an organic solvent, purifying optionally and oxidizing the resultant to oxidized coenzyme Q 10 .  
       
     
     
         34 . (Canceled)  
     
     
         35 . The process according to  claim 33 , 
 wherein the culture is carried out at 15 to 45° C. and at a pH of 4 to 9.    
     
     
         36 . The process according to  claim 33 , 
 wherein the concentration of the carbon source in the culture is controlled to a concentration that no adverse effects are substantially caused on the productivity of reduced coenzyme Q 10 .    
     
     
         37 . (Canceled)  
     
     
         38 . (Canceled)  
     
     
         39 . (Canceled)  
     
     
         40 . (Canceled)  
     
     
         41 . (Canceled)  
     
     
         42 . (Canceled)  
     
     
         43 . (Canceled)  
     
     
         44 . The process according to  claim 33 , 
 wherein the extraction of coenzymes Q 10  is carried out from wet cells or dry cells of the microbial cells or disrupted product thereof by using a hydrophilic organic solvent.    
     
     
         45 . (Canceled)  
     
     
         46 . The process according to  claim 33 , 
 wherein the extraction of the coenzymes Q 10  is carried out from an aqueous suspension of the microbial cells or disrupted product thereof by using a hydrophobic organic solvent.    
     
     
         47 . (Canceled)  
     
     
         48 . (Canceled)  
     
     
         49 . (Canceled)  
     
     
         50 . (Canceled)  
     
     
         51 . (Canceled)  
     
     
         52 . (Canceled)  
     
     
         53 . (Canceled)  
     
     
         54 . The process according to  claim 33 , 
 wherein the reduced coenzyme Q 10  is contained at a ratio of not less than 70 mole % among the entire coenzymes Q 10      in the case that the reduced coenzyme Q 10 -producing microorganisms are cultured with shaking (amplitude: 2 cm, 310 reciprocation/min) at 25° C. for 72 hours in 10 mL of a culture medium [(glucose: 20 g, peptone: 5 g, yeast extract: 3 g, malt extract: 3 g)/L, pH: 6.0] using a test tube (inner diameter: 21 mm, entire length: 200 mm),    the obtained broth is optionally concentrated,    the obtained solution is vigorously shaken for 3 minutes using 10 parts by volume of glass beads (425 to 600 μm) to disrupt the microorganisms under a nitrogen atmosphere in the concomitant presence of 3 parts by volume of isopropanol and 18.5 parts by volume of n-hexane relative to 10 parts by volume of the broth, and    the prepared hydrophobic organic solvent phase (n-hexane phase) is analyzed by HPLC.    
     
     
         55 . (Canceled)  
     
     
         56 . The process according to  claim 33 , 
 wherein the microorganisms are microorganisms of the genus  Agrobacterium,  the genus  Aspergillus,  the genus  Acetobacter,  the genus  Aminobacter,  the genus  Agromonas,  the genus  Acidiphilium,  the genus  Bulleromyces,  the genus  Bullera,  the genus  Brevundimonas,  the genus  Cryptococcus,  the genus  Chionosphaera,  the genus  Candida,  the genus  Cerinosterus,  the genus  Exisophiala,  the genus  Exobasidium,  the genus  Fellomyces,  the genus  Filobasidiella,  the genus  Filobasidium,  the genus  Geotrichum,  the genus  Graphiola,  the genus  Gluconobacter,  the genus  Kockovaella,  the genus  Kurtzmanomyces,  the genus  Lalaria,  the genus  Leucosporidium,  the genus  Legionella,  the genus  Methylobacterium,  the genus  Mycoplana,  the genus  Oosporidium,  the genus  Pseudomonas,  the genus  Psedozyma,  the genus  Paracoccus,  the genus  petromyces,  the genus  Rhodotorula,  the genus  Rhodosporidium,  the genus  Rhizomonas,  the genus  Rhodobium,  the genus  Rhodoplanes,  the genus  Rhodopseudomonas,  the genus  Rhodobacter,  the genus  Sporobolomyces,  the genus  Sporidiobolus,  the genus  Saitoella,  the genus  Schizosaccharomyces,  the genus  Sphingomonas,  the genus  Sporotrichum,  the genus  Sympodiomycopsis,  the genus  Sterigmatosporidium,  the genus  Tapharina,  the genus  Tremella,  the genus  Trichosporon,  the genus  Tilletiaria,  the genus  Tilletia,  the genus  Tolyposporium,  the genus  Tilletiopsis,  the genus  Ustilago,  the genus  Udeniomyces,  the genus  Xanthophilomyces,  the genus  Xanthobacter,  the genus  Paecilomyces,  the genus  Acremonium,  the genus  Hyhomonus,  or the genus  Rhizobium.      
     
     
         57 . (Canceled)