Heterologous Hosts

Abstract

This invention is related to bacterial engineering and the heterologous expression of useful compounds. In particular, the invention relates to a heterologous host that has been engineered for expression of a gene which is capable of polyketide or non-ribosomal peptide synthesis. Methods of treating cancer are also disclosed.

What is claimed: 
     
         1 . An  E. coli  Nissle 1917 host cell comprising a genetic engineering that provides heterologous expression of one or more genes capable of polyketide synthesis (PKS), non-ribosomal peptide synthesis (NRPS), or hybrid polyketide-NRP synthesis. 
     
     
         2 . The  E. coli  Nissle 1917 host cell according to  claim 1 , wherein the host cell is a heterologous species with respect to a species from which the one or more genes capable of polyketide, non-ribosomal peptide or hybrid polyketide-NRP synthesis is derived. 
     
     
         3 . The  E. coli  Nissle 1917 host cell according to  claim 1 , wherein the polyketide, NRP, or hybrid polyketide-NRP synthesis is not naturally produced in said host cell. 
     
     
         4 . The  E. coli  Nissle 1917 host cell according to  claim 1 , wherein the genetic engineering provides heterologous expression of all members of a gene cluster capable of polyketide, non-ribosomal peptide (NRP), or hybrid polyketide-NRP synthesis. 
     
     
         5 . The  E. coli  Nissle 1917 host cell according to  claim 1 , wherein the genetic engineering comprises transformation of  E. coli  Nissle 1917 with a gene or gene cluster from one or more organisms selected from the group consisting of myxobacteria,  Agrobacterium tumefaciens, Burkholderia  and  Stigmatella aurantiaca.    
     
     
         6 . The  E. coli  Nissle 1917 host cell according to  claim 5 , wherein the myxobacteria is selected from  Polyangium  and  Sorangium cellulosum.    
     
     
         7 . The  E. coli  Nissle 1917 host cell according to  claim 4 , wherein the gene cluster is a glidobactin gene cluster, a epothilone gene cluster, a tubulysin gene cluster, a disorazol(e) gene cluster, a salinomycin gene cluster, a myxochromide S gene cluster, a myxothiazol gene cluster, or a combination of the genes comprising these gene clusters. 
     
     
         8 . The  E. coli  Nissle 1917 host cell according to  claim 7 , wherein the host cell expresses glidobactin A, epothilone, myxochromide, colibactin, tubulysin, disorazol(e) or a functional derivative thereof. 
     
     
         9 . The  E. coli  Nissle 1917 host cell according to  claim 1 , wherein the one or more genes are found in nature in the gene cluster. 
     
     
         10 . The  E. coli  Nissle 1917 host cell according to  claim 1 , wherein the gene cluster comprises a genetic engineering. 
     
     
         11 . The  E. coli  Nissle 1917 host cell according to  claim 10 , wherein the genetic engineering comprises inclusion of one or more genes or one or more domains of genes from a heterologous gene cluster in the gene cluster. 
     
     
         12 . The  E. coli  Nissle 1917 host cell according to  claim 1 , wherein the genetic engineering comprises transformation of  E. coli  Nissle 1917 with a nucleic acid encoding a pPant transferase. 
     
     
         13 . The  E. coli  Nissle 1917 host cell of  claim 12 , wherein the genetic engineering comprises transformation of  E. coli  Nissle 1917 with a nucleic acid encoding MtaA pPant transferase. 
     
     
         14 . The  E. coli  Nissle 1917 host cell of  claim 13 , wherein expression of MtaA pPant transferase is regulated by a tetracycline inducible promoter. 
     
     
         15 . A compound with a molecular weight selected from 572.2, 678.3 and 671.3 obtainable by expressing MtaA pPant transferase in  E. coli  Nissle 1917. 
     
     
         16 . A method for generating a  E. coli  Nissle 1917 host cell comprising transforming  E. coli  Nissle 1917 with a heterologous gene or gene cluster capable of polyketide, non-ribosomal peptide (NRP), or hybrid polyketide-NRP synthesis. 
     
     
         17 . A method of treatment comprising administering the  E. coli  Nissle 1917 host cell of  claim 1  to a subject. 
     
     
         18 . A method of treatment comprising administering the compound of  claim 15  to a subject. 
     
     
         19 . A method of treating cancer comprising administering the  E. coli  Nissle 1917 host cell of  claim 1  to a tumor. 
     
     
         20 . A method of treating or preventing cancer comprising administering the compound of  claim 15  to a subject.