Process for producing polyhydroxyalkanoates by utilizing microorganisms

Abstract

A microbial polyhydroxyalkanoate which comprises one or more of monomer units represented by Formula (1), where R is at least one selected from the group represented by any one of Formulas (2), (3) and (4); in Formula (2), R1 is selected from the group consisting of hydrogen atom (H), halogen atom, —CN, —NO 2 , —CF 3 , —C 2 F 5 and —C 3 F 7 , and q is an integer of 1 to 8; in Formula (3), R2 is selected from the group consisting of hydrogen atom (H), halogen atom, —CN, —NO 2 , —CF 3 , —C 2 F 5 and —C 3 F 7 , and r is an integer of 1 to 8; in Formula (4), R3 is selected from the group consisting of hydrogen atom (H), halogen atom, —CN, —NO 2 , —CF 3 , —C 2 F 5 and —C 3 F 7 , and s is an integer of 1 to 8. The production method is also disclosed.

What is claimed is:  
     
         1 . A polyhydroxyalkanoate comprising one or more of monomer units represented by Formula (1),  
       
         
           
           
               
               
           
         
         where R is at least one selected from the group represented by any one of Formulas (2), (3) and (4);  
         
           
             
             
                 
                 
             
           
         
         in formula (2), R1 is selected from the group consisting of hydrogen atom (H), halogen atom, —CN, —NO 2 , —CF 3 , —C 2 F 5  and —C 3 F 7 , and q is an integer of 1 to 8;  
         in formula (3), R2 is selected from the group consisting of hydrogen atom (h), halogen atom, —CN, —NO 2 , —CF 3 , —C 2 F 5  and —C 3 F 7 , and r is an integer of 1 to 8;  
         in formula (4), R3 is selected from the group consisting of hydrogen atom (h), halogen atom, —CN, —NO 2 , —CF 3 , —C 2 F 5  and —C 3 F 7 , and s is an integer of 1 to 8;  
         provided that following R is not selected: 
 when one kind of R is selected: 
 R being R1=H and q=2, R being R1=H and q=3 in Formula (2),  
 R being R2=halogen and r=2, or R being R2=—CN and r=3 and  
 R being R2=—NO 2  and r=3 in Formula (3);  
 
 when two kinds of R are selected: 
 a combination of R being R1=H and q=3 and 5 respectively in Formula (2),  
 a combination of R being R2=H and r=2 and 4 respectively,  
 a combination of R being R2=H and r=2 and 6 respectively, and  
 a combination of R being R2=halogen and r=2 and 4 respectively in Formula (3);  
 
 when three kinds of R are selected: 
 a combination of R being R1=H and q=3, 5 and 7 respectively in Formula (2),  
 a combination of R being R2=H and r=1, 3 and 5 respectively, and a combination of R being R2=H and r=2, 4 and 6 respectively in Formula (3).  
 
 
       
     
     
         2 . The polyhydroxyalkanoate according to  claim 1  wherein the monomer unit is 3-hydroxy-4-phenoxybutyric acid unit of Formula (5).  
       
         
           
           
               
               
           
         
       
     
     
         3 . The polyhydroxyalkanoate according to  claim 1  wherein the monomer unit is 3-hydroxy-5-phenoxyvaleric acid unit of Formula (6).  
       
         
           
           
               
               
           
         
       
     
     
         4 . The polyhydroxyalkanoate according to  claim 1  wherein the monomer unit is 3-hydroxy-5-(4-fluorophenyl)valeric acid unit of Formula (7).  
       
         
           
           
               
               
           
         
       
     
     
         5 . The polyhydroxyalkanoate according to  claim 1  wherein the monomer unit is 3-hydroxy-4-cyclohexylbutyric acid unit of Formula (8).  
       
         
           
           
               
               
           
         
       
     
     
         6 . The polyhydroxyalkanoate according to  claim 1  wherein the monomer unit is 3-hydroxy-5-phenoxyvaleric acid unit of Formula (6) and 3-hydroxy-5-phenylvaleric acid unit of Formula (9).  
       
         
           
           
               
               
           
         
       
     
     
         7 . The polyhydroxyalkanoate according to  claim 1  wherein the monomer unit is 3-hydroxy-4-phenylbutyric acid unit of Formula (10) and 3-hydroxy-6-phenylhexanoic acid units of Formula (11).  
       
         
           
           
               
               
           
         
       
     
     
         8 . The polyhydroxyalkanoate according to any one of  claims 1  to  7  wherein a number average molecular weight is 10 to 200 thousands.  
     
     
         9 . A process of producing a polyhydroxyalkanoate comprising the step of: 
 culturing a microorganism in a culture medium containing a raw material alkanoate and an yeast extract, wherein the microorganism produces a polyhydroxyalkanoate utilizing the alkanoate.    
     
     
         10 . The process according to  claim 9 , wherein the alkanoate is represented by Formula (12), and the polyhydroxyalkanoate has one or more of monomer units represented by Formula (13):  
       
         
           
           
               
               
           
         
         wherein in Formula (12), R is at least one selected from the group consisting of Formulas (2), (3) and (4);  
         
           
             
             
                 
                 
             
           
         
         wherein in Formula (2), R1 is selected from the group consisting of hydrogen atom (H), halogen atom, —CN, —NO 2 , —CF 3 , —C 2 F 5  and —C 3 F 7 , and q is an integer of 1 to 8;  
         in Formula (3), R2 is selected from the group consisting of hydrogen atom (H), halogen atom, —CN, —NO 2 , —CF 3 , —C 2 F 5  and —C 3 F 7 , and r is an integer of 1 to 8;  
         in Formula (4), R3 is selected from the group consisting of hydrogen atom (H), halogen atom, —CN, —NO 2 , —CF 3 , —C 2 F 5  and —C 3 F 7 , and s is an integer of 1 to 8;  
         
           
             
             
                 
                 
             
           
         
         wherein R′ is the selected R for the alkanoate or a group having the same R1, R2 or R3 as R but q=q 0 , q=q 0 −2, q=q 0 −4 q=q 0 −6; or r=r 0 , r=r 0 −2, r=r 0 −4 or r=r 0 −6; or s=s 0  or s=s 0 −2, s=s 0 −4 or s=s 0 −6; wherein q 0 , r 0 , and s 0  are q, r, or s of the R, and q 0 −2, r 0 −2; s 0 −2, q 0 −4, r 0 −4; s 0 −4, q 0 −6, r 0 −6; or s 0 −6 being integers only more than 1.  
       
     
     
         11 . The process according to  claim 10  wherein the monomer unit is of Formula (5), and the microorganism is cultured in a culture medium containing 4-phenoxybutyric acid of Formula (14) and an yeast extract, the microorganism produces poly-3-hydroxy-4-phenoxybutyric acid utilizing 4-phenoxybutyric acid.  
       
         
           
           
               
               
           
         
       
     
     
         12 . The process according to  claim 10  wherein the monomer unit is of Formula (6), and the culture medium contains 5-phenoxyvaleric acid of Formula (15) and an yeast extract, and the microorganism produces poly-3-hydroxy-5-phenoxyvaleric acid utilizing 5-phenoxyvaleric acid.  
       
         
           
           
               
               
           
         
       
     
     
         13 . The process according to  claim 10  wherein the monomer unit is of Formula (16), the culture medium contains 5-(4-fluorophenoxy)valeric acid of Formula (17) and yeast extract, and the microorganism produces poly-3-hydroxy-5-(4-fluorophenoxy)valeric acid utilizing 5-(4-fluorophenoxy)valeric acid.  
       
         
           
           
               
               
           
         
       
     
     
         14 . The process according to  claim 10  wherein the monomer unit is of Formula (9), the culture medium contains 5-phenylvaleric acid of Formula (18) and an yeast extract, and the microorganism produces poly-3-hydroxy-5-phenylvaleric acid utilizing 5-phenylvaleric acid.  
       
         
           
           
               
               
           
         
       
     
     
         15 . The process according to  claim 10  wherein the monomer unit is of Formula (7), the culture medium contains 5-(4-fluorophenyl)valeric acid of Formula (19) and an yeast extract, and the microorganism produces poly-3-hydroxy-5-(4-fluorophenyl)valeric acid utilizing 5-(4-fluorophenyl)valeric acid.  
       
         
           
           
               
               
           
         
       
     
     
         16 . The process according to  claim 10  wherein the monomer unit is of Formula (8), the culture medium contains 4-cyclohexylbutyric acid of Formula (20) and an yeast extract, and the microorganism produces poly-3-hydroxy-4-cyclohexylbutyric acid utilizing 4-cyclohexylbutyric acid.  
       
         
           
           
               
               
           
         
       
     
     
         17 . The process according to  claim 10  wherein the monomer unit is of Formulas (6) and (9), the culture medium contains 5-phenoxyvaleric acid of Formula (15), 5-phenylvaleric acid of Formula (18) and an yeast extract, and the microorganism produces a polyhydroxyalkanoate comprising 3-hydroxy-5-phenoxyvaleric acid and 3-hydroxy-5-phenylvaleric acid utilizing 5-phenoxyvaleric acid and 5-phenylvaleric acid.  
       
         
           
           
               
               
           
         
       
     
     
         18 . The process according to  claim 10  wherein the monomer unit is of Formulas (10) and (11), the culture medium contains 6-phenylhexanoic acid of Formula (21) and an yeast extract, and the microorganism produces a polyhydroxyalkanoate comprising 3-hydroxy-4-phenylbutyric acid and 3-hydroxy-6-phenylhexanoic acid utilizing 6-phenylhexanoic acid.  
       
         
           
           
               
               
           
         
       
     
     
         19 . The process according to  claim 10  wherein the monomer unit is of Formulas (6) and (22), the culture medium contains 7-phenoxyheptanoic acid of Formula (23) and an yeast extract, and the microorganism produces a polyhydroxyalkanoate comprising 3-hydroxy-5-phenoxyvaleric acid and 3-hydroxy-7-phenoxyheptanoic acid utilizing 7-phenoxyheptanoic acid.  
       
         
           
           
               
               
           
         
       
     
     
         20 . The process according to  claim 10  wherein the monomer unit is of Formulas (5), (24) and (25), the culture medium contains 8-phenoxyoctanoic acid of Formula (26) and an yeast extract, and the microorganism produces a polyhydroxyalkanoate comprising 3-hydroxy-4-phenoxybutyric acid, 3-hydroxy-6-phenoxyhexanoic acid and 3-hydroxy-8-phenoxyoctanoic acid utilizing 8-phenoxyoctanoic acid.  
       
         
           
           
               
               
           
         
       
     
     
         21 . The process according to  claim 10  wherein the monomer unit is of Formulas (6), (22) and (27), the culture medium contains 11-phenoxyundecanoic acid of Formula (28) and an yeast extract, the microorganism produces a polyhydroxyalkanoate comprising 3-hydroxy-5-phenoxyvaleric acid, 3-hydroxy-7-phenoxyheptanoic acid and 3-hydroxy-9-phenoxynonanoic acid utilizing 11-phenoxyundecanoic acid.  
       
         
           
           
               
               
           
         
       
     
     
         22 . The process according to  claim 9  wherein the step of culturing is one step culture in a culture medium containing an alkanoate and an yeast extract.  
     
     
         23 . The process according to  claim 9  wherein the step of culturing is two step culture in a culture medium containing an alkanoate and an yeast extract followed by culturing in a nitrogen-restricted culture medium containing the alkanoate.  
     
     
         24 . The process according to  claim 9  which further comprises a step of separation/refinement of the polyhydroxyalkanoate.  
     
     
         25 . The process according to  claim 9  wherein the microorganism belongs to a genus Pseudomonas.  
     
     
         26 . The process according to  claim 25  wherein the microorganism is at least one selected from the group consisting of  Pseudomonas cichorii  YN2 (FERM BP-7375);  Pseudomonas cichorii  H45 (FERM BP-7374);  Pseudomonas putida  P91 (FERM BP-7373); and  Pseudomonas jessenii  P161 (FERM BP-7376).