Engineering complex microbial phenotypes with transcription enhancement
The present invention relates to a recombinant E. coli exhibiting a complex phenotype, comprising one or more RNA polymerase subunit genes, one or more functional genes, and, optionally, one or more transcription factors from a heterologous prokaryote. Also provided are methods for screening such a recombinant E. coli.
1. A method for generating a recombinant host E.coli bacterium exhibiting a desirable complex phenotype, comprising
(a) introducing into the host E. coli bacterium one or more RNA polymerase subunit genes from a heterologous prokaryote in a genus selected from the group consisting of Bacillus and Lactobacillus,
( b) expressing the one or more RNA poly merase subunit genes in the host E. coli bacterium;
(c) introducing into the host E. coli bacterium a plurality of function genes from one or more genomic libraries, and
(d) expressing the plurality of functional genes in the host E. coli bacterium, wherein the expression of the plurality of functional genese is enhanced by the expression of the one or more RNA polymerase subunit genes, whereby the host E. coli bacterium acquires the desirable complex phenotype.
2. The method of claim 1 , further comprising expressing one or more transcript ion factor genes in the host E.coli bacterium, wherein the one or more transcription factor genes are derived from the heterologous prokaryote.
3. The method of claim 1 , wherein the one or more RNA polymerase subunit genes comprise an alpha, beta, beta', omega or sigma subunit gene.
4. The method of claim 1 , wherein the one or more RNA polymerase subunit genes comprise a sigma subunit gene selected from the group consisting of rpoD, rpoN, sigH, rpoS, rpoE, sigB, sigF, sigE, and sigG.
5. The method of claim 1 , wherein the one or more genomic libraries are natural or enriched metagenomic libraries.
6. (Previousiy Presented) The method of ciaim 1 , wherein the one or more genomic libraries are generated from one or more heterologous prokaryotes.
7. The method of claim 1 , wherein the heterologous prokaryote is Lactobacillus plantarum or Bacills subtilis.
8. The method of claim 1 , wherein the one or more RNA polymerase subunit genes are selected from the group consisting of rpoD of Lactobacillus plantarun, sigA of Bacillus subtilis , and a combination thereof.