Genetic improvement of Escherichia coli for ethanol production: Chromosomal integration of Zymomonas mobilis genes encoding pyruvate decarboxylase and alcohol dehydrogenase II

Journal Article · Mon Apr 01 04:00:00 EST 1991 · Applied and Environmental Microbiology; (United States)

OSTI ID:5670592

Ohta, Kazuyoshi; Beall, D S; Mejia, J P; Shanmugam, K T; Ingram, L O ^[1]

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

Zymomonas mobilis genes for pyruvate decarboxylase (pdc) and alcohol dehydrogenase II (adhB) were integrated into the Escherichia coli chromosome within or near the pyruvate formate-lyase gene (pfl). Integration improved the stability of the Z. mobilis genes in E. coli, but further selection was required to increase expression. Spontaneous mutants were selected for resistance to high levels of chloramphenicol that also expressed high levels of the Z. mobilis genes. Analogous mutants were selected for increased expression of alcohol dehydrogenase on aldehyde indicator plates. These mutants were functionally equivalent to the previous plasmid-based strains for the fermentation of xylose and glucose to ethanol. Ethanol concentrations of 54.4 and 41.6 g/liter were obtained from 10% glucose and 8% xylose, respectively. The efficiency of conversion exceeded theoretical limits (0.51 g of ethanol/g of sugar) on the basis of added sugars because of the additional production of ethanol from the catabolism of complex nutrients. Further mutations were introduced to inactivate succinate production (frd) and to block homologous recombination (recA).

• OSTI ID: 5670592
• Journal Information: Applied and Environmental Microbiology; (United States), Journal Name: Applied and Environmental Microbiology; (United States) Vol. 57:4; ISSN 0099-2240; ISSN AEMID
• Country of Publication: United States
• Language: English

Related Subjects

09 BIOMASS FUELS
090900* -- Biomass Fuels-- Processing-- (1990-)
ALCOHOL DEHYDROGENASE
ALCOHOLS
ALDEHYDES
BACTERIA
BIOCONVERSION
BIOSYNTHESIS
BIOTECHNOLOGY
CARBOHYDRATES
CARBON-CARBON LYASES
CARBOXY-LYASES
DECARBOXYLASES
DNA
ENZYMES
ESCHERICHIA COLI
ETHANOL
FERMENTATION
GENES
GENETIC ENGINEERING
GLUCOSE
HEMIACETAL DEHYDROGENASES
HEXOSES
HYDROXY COMPOUNDS
LYASES
MICROORGANISMS
MONOSACCHARIDES
NUCLEIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
PENTOSES
PROTEINS
RECOMBINANT DNA
SACCHARIDES
SYNTHESIS
XYLOSE
ZYMOMONAS MOBILIS