Zymomonas pentose-sugar fermenting strains and uses thereof

Zhang, Min; Chou, Yat-Chen; Howe, William; Eddy, Christine; Evans, Kent; Mohagheghi, Ali

Title: Zymomonas pentose-sugar fermenting strains and uses thereof

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Abstract

Disclosed in the present invention is a Zymomonas integrant and derivatives of these integrants that posses the ability to ferment pentose into ethanol. The genetic sequences encoding for the pentose-fermenting enzymes are integrated into the Zymomonas in a two-integration event of homologous recombination and transposition. Each operon includes more than one pentose-reducing enzyme encoding sequence. The integrant in some embodiments includes enzyme sequences encoding xylose isomerase, xylulokinase, transketolase and transketolase. The Zymomonas integrants are highly stable, and retain activity for producing the pentose-fermenting enzyme for between 80 to 160 generations. The integrants are also resistant to acetate inhibition, as the integrants demonstrate efficient ethanol production even in the presence of 8 up to 16 grams acetate per liter media. These stably integrated sequences provide a unique Zymomonas that may then be used for the efficient conversion of pentose sugars (xylose, arabinose) to ethanol. Method of using the Zymomonas integrants and derivatives thereof in production of ethanol from cellulosic feedstock is also disclosed. The invention also provides a method for preparing a Zymomonas integrant as part of the present invention. The host Zymomonas strain found particularly useful in the creation of these compositions and methods is Zymomonas mobilis 31821.

Inventors:

Zhang, Min ^[1]; Chou, Yat-Chen ^[2]; Howe, William ^[2]; Eddy, Christine ^[2]; Evans, Kent ^[3]; Mohagheghi, Ali ^[4]

Lakewood, CO
Golden, CO
Littleton, CO
Northglenn, CO

Issue Date:: 2007-05-29

Research Org.:: Midwest Research Institute, Kansas City, MO (United States)

Sponsoring Org.:: USDOE

OSTI Identifier:: 909424

Patent Number(s):: 7223575

Assignee:: Midwest Research Institute (Kansas City, MO)

Patent Classifications (CPCs):: 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 {

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

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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}

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
C12N15/74 - Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
C12N9/1022 - {transferring aldehyde or ketonic groups

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

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
C12N15/902 - {using homologous recombination}

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 {
C12P7/06 - Ethanol, i.e. non-beverage

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
C12N9/92 - Glucose isomerase {
C12N15/90 - Stable introduction of foreign DNA into chromosome
C12N15/52 - Genes encoding for enzymes or proenzymes
C12N9/1205 - {Phosphotransferases with an alcohol group as acceptor

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DOE Contract Number:: AC36-99GO10337

Resource Type:: Patent

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES

Citation Formats


                    Zhang, Min, Chou, Yat-Chen, Howe, William, Eddy, Christine, Evans, Kent, and Mohagheghi, Ali. Zymomonas pentose-sugar fermenting strains and uses thereof.  United States: N. p., 2007. 
        Web.

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                    Zhang, Min, Chou, Yat-Chen, Howe, William, Eddy, Christine, Evans, Kent, & Mohagheghi, Ali. Zymomonas pentose-sugar fermenting strains and uses thereof.  United States.

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                    Zhang, Min, Chou, Yat-Chen, Howe, William, Eddy, Christine, Evans, Kent, and Mohagheghi, Ali. Tue .  
        "Zymomonas pentose-sugar fermenting strains and uses thereof".  United States.  https://www.osti.gov/servlets/purl/909424.

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@article{osti_909424,

  title        = {Zymomonas pentose-sugar fermenting strains and uses thereof},

  author       = {Zhang, Min and Chou, Yat-Chen and Howe, William and Eddy, Christine and Evans, Kent and Mohagheghi, Ali},

  abstractNote = {Disclosed in the present invention is a Zymomonas integrant and derivatives of these integrants that posses the ability to ferment pentose into ethanol. The genetic sequences encoding for the pentose-fermenting enzymes are integrated into the Zymomonas in a two-integration event of homologous recombination and transposition. Each operon includes more than one pentose-reducing enzyme encoding sequence. The integrant in some embodiments includes enzyme sequences encoding xylose isomerase, xylulokinase, transketolase and transketolase. The Zymomonas integrants are highly stable, and retain activity for producing the pentose-fermenting enzyme for between 80 to 160 generations. The integrants are also resistant to acetate inhibition, as the integrants demonstrate efficient ethanol production even in the presence of 8 up to 16 grams acetate per liter media. These stably integrated sequences provide a unique Zymomonas that may then be used for the efficient conversion of pentose sugars (xylose, arabinose) to ethanol. Method of using the Zymomonas integrants and derivatives thereof in production of ethanol from cellulosic feedstock is also disclosed. The invention also provides a method for preparing a Zymomonas integrant as part of the present invention. The host Zymomonas strain found particularly useful in the creation of these compositions and methods is Zymomonas mobilis 31821.},

  doi          = {},

  journal      = {},
number       = ,

  volume       = ,

  place        = {United States},

  year         = {2007},

  month        = {5}

}

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