Note: Sucrose transport and metabolism in Clostridium beijerinckii NCIMB 8052

Tangney, Martin; Rousse, Céline; Yazdanian, Mohsen; Mitchell, Wilfrid J.

doi:10.1046/j.1365-2672.1998.00432.x

Cited by 35 publications

(40 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…1), of which six have been assigned a role or putative function. Genome analysis reveals a similar organization in other bacteria, including several gram-positive organisms (5,30), with homologues of the transport/metabolic genes (gutAEBD) and the transcriptional activator gene (gutM) being highly conserved. However, GutQ has a narrow phylogenetic distribution confined to a subset of enterobacteria thus far limited to the sequenced gut operons in Escherichia, Shigella, and Salmonella species and Erwinia amylovora (1).…”

Section: Discussionmentioning

confidence: 99%

Identification of GutQ from Escherichia coli as a d -Arabinose 5-Phosphate Isomerase

Meredith¹,

Woodard

2005

J Bacteriol

View full text Add to dashboard Cite

The glucitol operon (gutAEBDMRQ) of Escherichia coli encodes a phosphoenolpyruvate:sugar phosphotransferase system that metabolizes the hexitol D-glucitol (sorbitol). The functions for all but the last gene, gutQ, have been previously assigned. The high sequence similarity between GutQ and KdsD, a D-arabinose 5-phosphate isomerase (API) from the 3-deoxy-D-manno-octulosonate (KDO)-lipopolysaccharide (LPS) biosynthetic pathway, suggested a putative activity, but its role within the context of the gut operon remained unclear. Accordingly, the enzyme was cloned, overexpressed, and characterized. Recombinant GutQ was shown to indeed be a second copy of API from the E. coli K-12 genome with biochemical properties similar to those of KdsD, catalyzing the reversible aldol-ketol isomerization between D-ribulose 5-phosphate (Ru5P) and Darabinose 5-phosphate (A5P). Genomic disruptions of each API gene were constructed in E. coli K-12. TCM11[(⌬kdsD)] was capable of sustaining essential LPS synthesis at wild-type levels, indicating that GutQ functions as an API inside the cell. The gut operon remained inducible in TCM7[(⌬gutQ)], suggesting that GutQ is not directly involved in D-glucitol catabolism. The conditional mutant TCM15[(⌬gutQ⌬kdsD)] was dependent on exogenous A5P both for LPS synthesis/growth and for upregulation of the gut operon. The phenotype was suppressed by complementation in trans with a plasmid encoding a functional copy of GutQ or by increasing the amount of A5P in the medium. As there is no obvious obligatory role for GutQ in the metabolism of D-glucitol and there is no readily apparent link between D-glucitol metabolism and LPS biosynthesis, it is suggested that A5P is not only a building block for KDO biosynthesis but also may be a regulatory molecule involved in expression of the gut operon.D-Arabinose 5-phosphate isomerase (API) is the first enzyme in the biosynthetic pathway of 3-deoxy-D-manno-octulosonate (KDO), a unique 8-carbon sugar component of lipopolysaccharides (LPSs). Lipopolysaccharides are amphiphilic macromolecules located in the outer leaflet of the outer membrane of gram-negative bacteria, forming an asymmetric lipid bilayer that surrounds the cell. The LPS layer is anchored in the outer membrane by the highly conserved lipid A domain and is followed by an oligosaccharide core region usually containing at least one KDO molecule that connects lipid A to an extended hypervariable repeating polysaccharide chain (O antigen) (25). In addition to providing a measure of intrinsic antibiotic resistance and defense against host responses, LPS (also referred to as endotoxin) is the main mediator of gramnegative bacterial pathogenesis (13, 33). The minimal LPS structure required for viability of Escherichia coli under laboratory conditions is KDO 2 -lipid A, also referred to as Re endotoxin (11,25,31).API catalyzes the reversible 1,2-keto/aldol isomerization of the pentose pathway intermediate ribulose 5-phosphate (Ru5P) to D-arabinose 5-phosphate (A5P) and is the intracellular source of A5P used f...

show abstract

Section: Discussionmentioning

confidence: 99%

Identification of GutQ from Escherichia coli as a d -Arabinose 5-Phosphate Isomerase

Meredith¹,

Woodard

2005

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…In E. coli, another sequence is present (talC, see above) which shared 68% identical (79% similar) residues with FSA. fsa-related genes with prominent similarity were only found in prokaryotic genomes such as in Clostridium beijerinckii (31), as well as in the total genomes of Yersinia pestis, Bacillus subtilis, and Bacillus stearothermophilus, in the extreme thermophilic eubacteria Aquifex aeolicus and Thermotoga maritima, and in the archaebacterium Methanococcus jannaschii. Fig.…”

Section: Fructose-6-phosphate Aldolase From E Colimentioning

confidence: 99%

Fructose-6-phosphate Aldolase Is a Novel Class I Aldolase from Escherichia coli and Is Related to a Novel Group of Bacterial Transaldolases

Schürmann

Sprenger

2001

Journal of Biological Chemistry

193

171

View full text Add to dashboard Cite

We have cloned an open reading frame from the Escherichia coli K-12 chromosome that had been assumed earlier to be a transaldolase or a transaldolase-related protein, termed MipB. Here we show that instead a novel enzyme activity, fructose-6-phosphate aldolase, is encoded by this open reading frame, which is the first report of an enzyme that catalyzes an aldol cleavage of fructose 6-phosphate from any organism. We propose the name FSA (for fructose-six phosphate aldolase; gene name fsa). The recombinant protein was purified to apparent homogeneity by anion exchange and gel permeation chromatography with a yield of 40 mg of protein from 1 liter of culture. By using electrospray tandem mass spectroscopy, a molecular weight of 22,998 per subunit was determined. From gel filtration a size of 257,000 (؎ 20,000) was calculated. The enzyme most likely forms either a decamer or dodecamer of identical subunits. The purified enzyme displayed a V max of 7 units mg ؊1 of protein for fructose 6-phosphate cleavage (at 30°C, pH 8.5 in 50 mM glycylglycine buffer). For the aldolization reaction a V max of 45 units mg ؊1 of protein was found; K m values for the substrates were 9 mM for fructose 6-phosphate, 35 mM for dihydroxyacetone, and 0.8 mM for glyceraldehyde 3-phosphate. FSA did not utilize fructose, fructose 1-phosphate, fructose 1,6-bisphosphate, or dihydroxyacetone phosphate. FSA is not inhibited by EDTA which points to a metal-independent mode of action. The lysine 85 residue is essential for its action as its exchange to arginine (K85R) resulted in complete loss of activity in line with the assumption that the reaction mechanism involves a Schiff base formation through this lysine residue (class I aldolase). Another fsa-related gene, talC of Escherichia coli, was shown to also encode fructose-6-phosphate aldolase activity and not a transaldolase as proposed earlier.

show abstract

“…Many bacterial species, including Klebsiella pneumoniae (Sprenger & Lengeler, 1988 ;Titgemeyer et al, 1996), Bacillus subtilis (Fouet et al, 1987), Lactococcus lactis (Thompson & Chassy, 1981 ;Thompson et al, 1991 ;Rauch & deVos, 1992), Fusobacterium mortiferum (Thompson et al, 1992), Escherichia coli (Schmid et al, 1988) and Clostridium beijerinckii (Tangney et al, 1998 ;Reid et al, 1999) with phosphorylation at C-6 of the glucosyl moiety via the phosphoenolpyruvate-dependent sucrose : phosphotransferase system (PEP : PTS) (Meadow et al, 1990 ;Postma et al, 1993). Sucrose 6-phosphate is hydrolysed intracellularly by sucrose-6-phosphate hydrolase (S6PH) to yield glucose 6-phosphate and fructose, which are further metabolized via the glycolytic pathway.…”

Section: Introductionmentioning

confidence: 99%

Metabolism of sucrose and its five isomers by Fusobacterium mortiferum

et al. 2002

View full text Add to dashboard Cite

Fusobacterium mortiferum utilizes sucrose [glucose-fructose in α(1 2) linkage] and its five isomeric α-D-glucosyl-D-fructoses as energy sources for growth.Sucrose-grown cells are induced for both sucrose-6-phosphate hydrolase (S6PH) and fructokinase (FK), but the two enzymes are not expressed above constitutive levels during growth on the isomeric compounds. Extracts of cells grown previously on the sucrose isomers trehalulose α(1 1), turanose α(1 3), maltulose α(1 4), leucrose α(1 5) and palatinose α(1 6) contained high levels of an NAD M plus metal-dependent phospho-α-glucosidase (MalH). The latter enzyme was not induced during growth on sucrose. MalH catalysed the hydrolysis of the 6'-phosphorylated derivatives of the five isomers to yield glucose 6-phosphate and fructose, but sucrose 6-phosphate itself was not a substrate. Unexpectedly, MalH hydrolysed both α-and β-linked stereomers of the chromogenic analogue p-nitrophenyl glucoside 6-phosphate. The gene malH is adjacent to malB and malR, which encode an EII(CB) component of the phosphoenolpyruvate-dependent sugar :phosphotransferase system and a putative regulatory protein, respectively. The authors suggest that for F. mortiferum, the products of malB and malH catalyse the phosphorylative translocation and intracellular hydrolysis of the five isomers of sucrose and of related α-linked glucosides. Genes homologous to malB and malH are present in both Klebsiella pneumoniae and the enterohaemorrhagic strain Escherichia coli O157 :H7. Both these organisms grew well on sucrose, but only K. pneumoniae exhibited growth on the isomeric compounds.

show abstract

Note: Sucrose transport and metabolism in Clostridium beijerinckii NCIMB 8052

Cited by 35 publications

References 22 publications

Identification of GutQ from Escherichia coli as a d -Arabinose 5-Phosphate Isomerase

Identification of GutQ from Escherichia coli as a d -Arabinose 5-Phosphate Isomerase

Fructose-6-phosphate Aldolase Is a Novel Class I Aldolase from Escherichia coli and Is Related to a Novel Group of Bacterial Transaldolases

Metabolism of sucrose and its five isomers by Fusobacterium mortiferum

Contact Info

Product

Resources

About