Properties of the glucose phosphotransferase system of Clostridium acetobutylicum NCIB 8052

Journal Article · Sun Sep 01 00:00:00 EDT 1991 · Applied and Environmental Microbiology; (United States)

OSTI ID:5910047

Mitchell, W J; Shaw, J E; Andrews, L ^[1]

---

1. Heriot-Watt Univ., Edinburgh (England)

Acetone-butanol-ethanol fermentation by Clostridium acetobutylicum has been exploited on an industrial scale in the past, but for economic reasons the process has declined. However, with an increased understanding of solvent formation and the potential for genetic manipulation of the organism, this fermentation is once again receiving attention. An economical process would be founded on the use of cheap, renewable substrates, ideally carbohydrate-based waste materials. However, little is known about the mechanism and regulation of carbohydrate accumulation by C. acetobutylicum. The glucose phosphotransferase system (PTS) of C. acetobutylicum was studied by using cell extracts. The system exhibited a K{sub m} for glucose of 34 {mu}M, and glucose phosphorylation was inhibited competitively by mannose and 2-deoxyglucose. The analogs 3-O-methylglucoside and methyl {alpha}-glucoside did not inhibit glucose phosphorylation significantly. Activity showed no dependence on Mg{sup 2+} ions or on pH in the range 6.0 to 8.0. The PTS comprised both soluble and membrane-bound proteins, which interacted functionally with the PTSs of Clostridium pasteurianum, Bacillus subtilis, and Escherichia coli. In addition to a membrane-bound enzyme II{sup Glc}, sugar phosphorylation assays in heterologous systems incorporating extracts of pts mutants of other organisms provided evidence for enzyme I, HPr, and III{sup Glc} components. The HPr was found in the soluble fraction of C. acetobutylicum extracts, whereas enzyme I, and probably also III{sup Glc}, was present in both the soluble and membrane fractions, suggesting a membrane location in the intact cell.

• OSTI ID: 5910047
• Journal Information: Applied and Environmental Microbiology; (United States), Vol. 57:9; ISSN 0099-2240
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
09 BIOMASS FUELS
CLOSTRIDIUM ACETOBUTYLICUM
METABOLISM
FUELS
BIOSYNTHESIS
GLUCOSE
BACILLUS SUBTILIS
CARBOHYDRATES
CATIONS
COMPARATIVE EVALUATIONS
ENZYME ACTIVITY
ESCHERICHIA COLI
INHIBITION
MAGNESIUM COMPOUNDS
PH VALUE
PHOSPHOTRANSFERASES
ALDEHYDES
ALKALINE EARTH METAL COMPOUNDS
BACILLUS
BACTERIA
CHARGED PARTICLES
CLOSTRIDIUM
ENZYMES
EVALUATION
HEXOSES
IONS
METHANOGENIC BACTERIA
MICROORGANISMS
MONOSACCHARIDES
ORGANIC COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PROTEINS
SACCHARIDES
SYNTHESIS
TRANSFERASES
550200* - Biochemistry
090900 - Biomass Fuels- Processing- (1990-)