Production of polyalkanoate

Abstract

Genes encoding polyhydroxyalkanoate synthase, β-ketothiolase and acetoacetyl CoA reductase are isolated from the publicly available bacterium Chromatium vinosum. Recombinant genomes of plants or other species of bacteria which contain these genes are capable of producing polyalkanoate polymers. The nucleotide sequences of the said three genes have been determined.

1 . Gene fragments isolated from the bacterium  Chromatium vinosum  and encoding polyhydroxyalkanoate (PHA) synthase, acetoacetyl CoA reductase and β-ketothiolase.  
     
     
         2 . Gene fragments as claimed in  claim 1  in which the  Chromatium vinosum  is of the strain designated D, available to the public from the Deutsche Sammlung fur Mikroorganismen under the Accession Number 180.  
     
     
         3 . A 16.5kb EcoRl fragment of  Chromatium vinosum  DNA, designated PP10, hybridizable to a 5.2 kb SmaI/EcoRl fragment, designated SE52 isolated from  Alcaligenes eutrophus  and known to contain the genes encoding PHA-synthase acetoacetyl CoA reductase and β-ketothiolase.  
     
     
         4 . A fragment of the PP10 fragment claimed in  claim 3 , designated SE45, encoding the PHA-synthase and β-ketothiolase genes.  
     
     
         5 . A fragment of the PP10 fragment claimed in  claim 3 , designated SB24, encoding the acetoacetyl CoA reductase gene.  
     
     
         6 . A recombinant genome which contains one or more of the fragments designated PP10, SE45 and region SB24 claimed in claims  3 ,  4  and  5  respectively.  
     
     
         7 . A bacterium having incorporated in its genome one or more of the fragments designated PP10, SE45 and region SB24 claimed in claims  3 ,  4  and  5  respectively.  
     
     
         8 . A plant having stably incorporated in its genome by transformation one or more of the fragments designated PP10, SE45 and region SB24 claimed in claims  3 ,  4  and  5  respectively.  
     
     
         9 . A method for the manufacture of polyhydroxyalkanoates, comprising culturing the microorganism  Chromatium vinosum,  or a bacterium of a different species having stably incorporated within its genome by transformation one or more PHA synthesising genes from  Chromatium vinosum.    
     
     
         10 . A gene, encoding β-ketothiolase, having the nucleotide sequence shown in FIG. 3.  
     
     
         11 . A gene encoding polyhydroxyalkanoate synthase (phbC), having the nucleotide sequence shown in FIG. 5.  
     
     
         12 . A gene encoding acetoacetyl CoA reductase (phbB) having the nucleotide sequence shown in FIG. 5.