Title: Recombinant cells that highly express chromosomally-integrated heterologous genes

Abstract

Recombinant host cells are obtained that comprise (A) a heterologous, polypeptide-encoding polynucleotide segment, stably integrated into a chromosome, which is under transcriptional control of an endogenous promoter and (B) a mutation that effects increased expression of the heterologous segment, resulting in enhanced production by the host cells of each polypeptide encoded by that segment, relative to production of each polypeptide by the host cells in the absence of the mutation. The increased expression thus achieved is retained in the absence of conditions that select for cells displaying such increased expression. When the integrated segment comprises, for example, ethanol-production genes from an efficient ethanol producer like Zymomonas mobilis, recombinant Escherichia coli and other enteric bacterial cells within the present invention are capable of converting a wide range of biomass-derived sugars efficiently to ethanol.

Inventors:

Ingram, Lonnie O ^[1]; Ohta, Kazuyoshi ^[1]; Wood, Brent E ^[1]

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1. Gainesville, FL

Issue Date:

Thu Jan 01 00:00:00 EST 1998

Research Org.:

Univ. of Florida, Gainesville, FL (United States)

OSTI Identifier:

871892

Patent Number(s):

5821093

Assignee:

University of Florida Research Foundation, Inc. (Gainesville, FL)

Patent Classifications (CPCs):

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C - CHEMISTRY C12 - BIOCHEMISTRY C12P - FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {

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C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C12N15/01 - Preparation of mutants without inserting foreign genetic material therein
C12N15/52 - Genes encoding for enzymes or proenzymes
C12N15/70 - Vectors or expression systems specially adapted for E. coli
C12N15/74 - Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
C12N15/902 - {using homologous recombination}
C12N9/0006 - {acting on CH-OH groups as donors
C12N9/88 - Lyases

C - CHEMISTRY C12 - BIOCHEMISTRY C12P - FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {

C12P7/065 - {with microorganisms other than yeasts}
C12P7/10 - substrate containing cellulosic material

G - PHYSICS G06 - COMPUTING G06T - IMAGE DATA PROCESSING OR GENERATION, IN GENERAL

G06T1/00 - General purpose image data processing

H - ELECTRICITY H04 - ELECTRIC COMMUNICATION TECHNIQUE H04N - PICTORIAL COMMUNICATION, e.g. TELEVISION

H04N1/00 - Scanning, transmission or reproduction of documents or the like, e.g. facsimile transmission

Y - NEW / CROSS SECTIONAL TECHNOLOGIES Y02 - TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE Y02E - REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

Y02E50/10 - Biofuels

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DOE Contract Number:  

FG05-86ER13574

Resource Type:

Patent

Country of Publication:

United States

Language:

English

Subject:

recombinant; cells; highly; express; chromosomally-integrated; heterologous; genes; host; obtained; comprise; polypeptide-encoding; polynucleotide; segment; stably; integrated; chromosome; transcriptional; control; endogenous; promoter; mutation; effects; increased; expression; resulting; enhanced; production; polypeptide; encoded; relative; absence; achieved; retained; conditions; select; displaying; comprises; example; ethanol-production; efficient; ethanol; producer; zymomonas; mobilis; escherichia; coli; enteric; bacterial; capable; converting; wide; range; biomass-derived; sugars; efficiently; zymomonas mobilis; wide range; host cells; recombinant host; host cell; recombinant cells; segment comprises; escherichia coli; heterologous genes; stably integrated; chromosomally-integrated heterologous; express chromosomally-integrated; highly express; polypeptide-encoding polynucleotide; polynucleotide segment; /435/999/