|Gene:||fbp||Accession Numbers: EG10283 (EcoCyc), b4232, ECK4227|
Subunit composition of
fructose-1,6-bisphosphatase I = [Fbp]4
fructose-1,6-bisphosphatase monomer = Fbp
Fructose-1,6-bisphosphatase catalyzes the conversion of fructose-1,6-bisphosphate to fructose-6-phosphate in the gluconeogenesis pathway. The enzyme is required for growth on glycerol, succinate and acetate as the carbon source, but not for growth on hexoses and pentoses [Fraenkel65].
To avoid futile cycling with phosphofructokinases, the activity of fructose-1,6-bisphosphatase must be regulated. Like the mammalian enzymes, E. coli fructose-1,6-bisphosphatase is inhibited by fructose-2,6-bisphosphate; however, this compound is not present in vivo [KelleyLoughnane02]. AMP is a noncompetitive inhibitor of the enzyme. The low Ki of the enzyme for AMP suggests that under physiological conditions the enzyme would be inhibited to a great extent [Babul83]. A mutant enzyme that is insensitive to AMP inhibition has been isolated [Sedivy86]. Phosphoenolpyruvate (PEP) was initially reported to partially inhibit fructose-1,6-bisphosphatase activity at high concentrations, but was able to block inhibition by AMP [Babul83]. A later report showed that PEP at low concentrations (2 mM) activates the enzyme; PEP may thus be the physiological regulator under gluconeogenic growth conditions [Hines06].
Under nondenaturing conditions, the enzyme is present in several aggregated forms in which the tetramer seems to predominate at low enzyme concentrations [Babul83]. A crystal structure of this enzyme has been solved at 1.45 Å resolution [Hines06]. Crystal structures have also been solved with several bound ligands at 2.05 Å resolution [Hines07] and 2.18 Å resolution [Hines07a, Hines07b].
In addition to PEP, citrate is also an activator. Tetramers with bound PEP or citrate are in the active allosteric R state conformation. PEP, citrate, 3-phosphoglycerate, cis-aconitate, isocitrate, oxaloacetate, and α-ketoglutarate are activators at concentrations of up to 5 mM, although PEP is the most potent activator. Like PEP, citrate is an antagonist of AMP inhibition [Hines07a]. Glucose 6-phosphate is also an allosteric inhibitor and distinct sites for inhibition by AMP and glucose 6-phosphate have been identified. The enzyme undergoes a quaternary structure transition from the R-state to a T-like state in response to the binding of these metabolites. Their binding is proposed to down-regulate the activated R state enzyme [Hines07]. Unlike the porcine enzyme which is synergistically inhibited by AMP and fructose 2,6-bisphosphate, these compounds are not synergistic inhibitors of this Type I E. coli enzyme [Hines07b, Gao14].
Using a kinetic model of E. coli glycolysis, allosteric interactions were identified that govern the reversible switch between glycolysis and gluconeogenesis which included the activation of fructose-1,6-bisphosphatase by pyruvate [Link13]. In E. coli metabolic engineering studies, site-directed mutagenesis of target amino acid residues encoded by fbp reduced allosteric inhibition without affecting catalytic efficiency [Yang12a]. E. coli fbp overexpressed in a strain of Corynebacterium glutamicum resulted in increased L-lysine production [Xu14].
A series of vectors inducibly expressing paired-terminus antisense RNAs was constructed to silence central carbon metabolism in host E. coli K-12 MG1655. A vector that silenced fbp at 77% efficacy resulted in a defect in carbon catabolite repression [Nakashima14].
|Map Position: [4,452,634 <- 4,453,632] (95.97 centisomes, 345°)||Length: 999 bp / 332 aa|
Molecular Weight of Polypeptide: 36.834 kD (from nucleotide sequence), 40 kD (experimental) [Babul83 ]
Molecular Weight of Multimer: 150 kD (experimental) [KelleyLoughnane02]
Unification Links: ASAP:ABE-0013842 , CGSC:784 , EchoBASE:EB0279 , EcoGene:EG10283 , EcoliWiki:b4232 , ModBase:P0A993 , OU-Microarray:b4232 , PortEco:fbp , PR:PRO_000022576 , Pride:P0A993 , Protein Model Portal:P0A993 , RefSeq:NP_418653 , RegulonDB:EG10283 , SMR:P0A993 , String:511145.b4232 , Swiss-Model:P0A993 , UniProt:P0A993
Relationship Links: InterPro:IN-FAMILY:IPR000146 , InterPro:IN-FAMILY:IPR020548 , InterPro:IN-FAMILY:IPR028343 , Panther:IN-FAMILY:PTHR11556 , PDB:Structure:2GQ1 , PDB:Structure:2OWZ , PDB:Structure:2OX3 , PDB:Structure:2Q8M , PDB:Structure:2QVR , Pfam:IN-FAMILY:PF00316 , Prints:IN-FAMILY:PR00115 , Prosite:IN-FAMILY:PS00124
|Biological Process:||GO:0006094 - gluconeogenesis
GO:0016311 - dephosphorylation [GOA06, GOA01, GOA01a, KelleyLoughnane02]
GO:0051289 - protein homotetramerization [KelleyLoughnane02]
GO:0005975 - carbohydrate metabolic process [UniProtGOA11a, GOA01a]
GO:0008152 - metabolic process [UniProtGOA11a]
GO:0016051 - carbohydrate biosynthetic process [GOA06]
|Molecular Function:||GO:0042132 - fructose 1,6-bisphosphate 1-phosphatase activity
[GOA06, GOA01, GOA01a, KelleyLoughnane02]
GO:0000166 - nucleotide binding [UniProtGOA11a]
GO:0000287 - magnesium ion binding [GOA06]
GO:0003824 - catalytic activity [UniProtGOA11a]
GO:0016787 - hydrolase activity [UniProtGOA11a]
GO:0042578 - phosphoric ester hydrolase activity [GOA01a]
GO:0046872 - metal ion binding [UniProtGOA11a]
|Cellular Component:||GO:0005829 - cytosol
[DiazMejia09, Ishihama08, LopezCampistrou05]
GO:0005737 - cytoplasm [UniProtGOA11, UniProtGOA11a, GOA06]
|MultiFun Terms:||metabolism → central intermediary metabolism|
|Growth Medium||Growth?||T (°C)||O2||pH||Osm/L||Growth Observations|
|LB enriched||Yes||37||Aerobic||6.95||Yes [Gerdes03, Comment 1]|
|LB Lennox||Yes||37||Aerobic||7||Yes [Baba06, Comment 2]|
|M9 medium with 1% glycerol||Yes||37||Aerobic||7.2||0.35||Yes [Joyce06, Comment 3]|
|MOPS medium with 0.4% glucose||Yes||37||Aerobic||7.2||0.22||Yes [Baba06, Comment 2] |
Yes [Feist07, Comment 4]
Enzymatic reaction of: fructose-1,6-bisphosphatase
Synonyms: fructose bisphosphatase, hexosediphosphatase, fructose-1,6-P2 1-phosphatase
EC Number: 184.108.40.206
The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction in which it was curated.
The reaction is physiologically favored in the direction shown.
In Pathways: superpathway of hexitol degradation (bacteria) , superpathway of glycolysis and Entner-Doudoroff , superpathway of glycolysis, pyruvate dehydrogenase, TCA, and glyoxylate bypass , gluconeogenesis I , glycolysis II (from fructose 6-phosphate) , glycolysis I (from glucose 6-phosphate)
The enzyme shows substrate inhibition by fructose1,6-bisphosphate at concentrations above 0.05mM [Babul83].
Values for kinetic parameters were shown to vary with assay protocol [Hines07a].
pH(opt): 7.5 [Babul83]
|Feature Class||Location||Attached Group||Citations||Comment|
|Protein-Segment||3 -> 5|
|Nucleotide-Phosphate-Binding-Region||19 -> 23||AMP|
|Nucleotide-Phosphate-Binding-Region||104 -> 105||AMP|
|Protein-Segment||113 -> 116|
|Protein-Segment||257 -> 259|
10/20/97 Gene b4232 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10283; confirmed by SwissProt match.
Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554
DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114
Feist07: Feist AM, Henry CS, Reed JL, Krummenacker M, Joyce AR, Karp PD, Broadbelt LJ, Hatzimanikatis V, Palsson BO (2007). "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information." Mol Syst Biol 3;121. PMID: 17593909
Gao14: Gao Y, Shen L, Honzatko RB (2014). "Central cavity of fructose-1,6-bisphosphatase and the evolution of AMP/fructose 2,6-bisphosphate synergism in eukaryotic organisms." J Biol Chem 289(12);8450-61. PMID: 24436333
Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938
Hines07: Hines JK, Kruesel CE, Fromm HJ, Honzatko RB (2007). "Structure of inhibited fructose-1,6-bisphosphatase from Escherichia coli: distinct allosteric inhibition sites for AMP and glucose 6-phosphate and the characterization of a gluconeogenic switch." J Biol Chem 282(34);24697-706. PMID: 17567577
Hines07a: Hines JK, Fromm HJ, Honzatko RB (2007). "Structures of activated fructose-1,6-bisphosphatase from Escherichia coli. Coordinate regulation of bacterial metabolism and the conservation of the R-state." J Biol Chem 282(16);11696-704. PMID: 17314096
Hines07b: Hines JK, Chen X, Nix JC, Fromm HJ, Honzatko RB (2007). "Structures of mammalian and bacterial fructose-1,6-bisphosphatase reveal the basis for synergism in AMP/fructose 2,6-bisphosphate inhibition." J Biol Chem 282(49);36121-31. PMID: 17933867
Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394
KelleyLoughnane02: Kelley-Loughnane N, Biolsi SA, Gibson KM, Lu G, Hehir MJ, Phelan P, Kantrowitz ER (2002). "Purification, kinetic studies, and homology model of Escherichia coli fructose-1,6-bisphosphatase." Biochim Biophys Acta 1594(1);6-16. PMID: 11825604
Link13: Link H, Kochanowski K, Sauer U (2013). "Systematic identification of allosteric protein-metabolite interactions that control enzyme activity in vivo." Nat Biotechnol 31(4);357-61. PMID: 23455438
LopezCampistrou05: Lopez-Campistrous A, Semchuk P, Burke L, Palmer-Stone T, Brokx SJ, Broderick G, Bottorff D, Bolch S, Weiner JH, Ellison MJ (2005). "Localization, annotation, and comparison of the Escherichia coli K-12 proteome under two states of growth." Mol Cell Proteomics 4(8);1205-9. PMID: 15911532
Marcus84: Marcus F, Edelstein I, Rittenhouse J (1984). "Inhibition of Escherichia coli fructose-1,6-bisphosphatase by fructose 2,6-bisphosphate." Biochem Biophys Res Commun 119(3);1103-8. PMID: 6324777
Nakashima14: Nakashima N, Ohno S, Yoshikawa K, Shimizu H, Tamura T (2014). "A vector library for silencing central carbon metabolism genes with antisense RNAs in Escherichia coli." Appl Environ Microbiol 80(2);564-73. PMID: 24212579
Xu14: Xu JZ, Zhang JL, Guo YF, Jia QD, Zhang WG (2014). "Heterologous expression of Escherichia coli fructose-1,6-bisphosphatase in Corynebacterium glutamicum and evaluating the effect on cell growth and L-lysine production." Prep Biochem Biotechnol 44(5);493-509. PMID: 24397720
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