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Escherichia coli K-12 substr. MG1655 Enzyme: fructose-1,6-bisphosphatase I



Gene: fbp Accession Numbers: EG10283 (EcoCyc), b4232, ECK4227

Synonyms: fdp

Regulation Summary Diagram: ?

Subunit composition of fructose-1,6-bisphosphatase I = [Fbp]4
         fructose-1,6-bisphosphatase monomer = Fbp

Summary:
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, 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 shows 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 the enzyme has been solved at 1.45 Å resolution [Hines06].

Locations: cytosol

Map Position: [4,452,634 <- 4,453,632] (95.97 centisomes)
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]

pI: 5.97

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

Gene-Reaction Schematic: ?

GO Terms:

Biological Process: GO:0006094 - gluconeogenesis Inferred from experiment Inferred by computational analysis [UniProtGOA12, Fraenkel65]
GO:0016311 - dephosphorylation Inferred by computational analysis Inferred from experiment [KelleyLoughnane02, GOA06, GOA01, GOA01a]
GO:0005975 - carbohydrate metabolic process Inferred by computational analysis [UniProtGOA11a, GOA01a]
GO:0008152 - metabolic process Inferred by computational analysis [UniProtGOA11a]
GO:0016051 - carbohydrate biosynthetic process Inferred by computational analysis [GOA06]
Molecular Function: GO:0042132 - fructose 1,6-bisphosphate 1-phosphatase activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01, GOA01a, KelleyLoughnane02]
GO:0000166 - nucleotide binding Inferred by computational analysis [UniProtGOA11a]
GO:0000287 - magnesium ion binding Inferred by computational analysis [GOA06]
GO:0003824 - catalytic activity Inferred by computational analysis [UniProtGOA11a]
GO:0016787 - hydrolase activity Inferred by computational analysis [UniProtGOA11a]
GO:0042578 - phosphoric ester hydrolase activity Inferred by computational analysis [GOA01a]
GO:0046872 - metal ion binding Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [DiazMejia09, Ishihama08, LopezCampistrou05]
GO:0005737 - cytoplasm Inferred by computational analysis [UniProtGOA11, UniProtGOA11a, GOA06]

MultiFun Terms: metabolism central intermediary metabolism

Essentiality data for fbp knockouts: ?

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]

Credits:
Last-Curated ? 20-Dec-2006 by Keseler I , SRI International


Enzymatic reaction of: fructose-1,6-bisphosphatase

Synonyms: fructose bisphosphatase, hexosediphosphatase, fructose-1,6-P2 1-phosphatase

EC Number: 3.1.3.11

fructose 1,6-bisphosphate + H2O <=> β-D-fructofuranose 6-phosphate + phosphate

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction of enzyme catalysis.

The reaction is physiologically favored in the direction shown.

Alternative Substrates for fructose 1,6-bisphosphate: α-glucose 1,6-bisphosphate [Babul83 ] , β-D-fructofuranose 1-phosphate [Babul83 ]

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-6P) , glycolysis I (from glucose-6P)

Summary:
The enzyme shows substrate inhibition by fructose1,6-bisphosphate at concentrations above 0.05mM [Babul83].

Cofactors or Prosthetic Groups: Mg2+ [KelleyLoughnane02, Babul83]

Activators (Unknown Mechanism): sulfate [Hines06] , phosphoenolpyruvate [Hines06]

Inhibitors (Competitive): β-D-fructose 2,6-bisphosphate [KelleyLoughnane02, Marcus84]

Inhibitors (Noncompetitive): AMP [Babul83]

Inhibitors (Unknown Mechanism): phosphoenolpyruvate [Babul83]

Primary Physiological Regulators of Enzyme Activity: phosphoenolpyruvate , AMP

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
fructose 1,6-bisphosphate
2.0
[Babul83, BRENDA14]
fructose 1,6-bisphosphate
15.4
[KelleyLoughnane02]
fructose 1,6-bisphosphate
100.0
2.5
[Brown09, BRENDA14]

pH(opt): 7.8 [BRENDA14, KelleyLoughnane02], 7.5 [Babul83]


Sequence Features

Feature Class Location Citations Comment
Protein-Segment 3 -> 5
[UniProt10]
UniProt: Allosteric activator binding; Sequence Annotation Type: region of interest;
Nucleotide-Phosphate-Binding-Region 19 -> 23
[UniProt10a]
UniProt: AMP;
Amino-Acid-Sites-That-Bind 30
[UniProt10a]
UniProt: Allosteric activator;
Metal-Binding-Site 89
[UniProt10a]
UniProt: Magnesium 1;
Nucleotide-Phosphate-Binding-Region 104 -> 105
[UniProt10a]
UniProt: AMP;
Metal-Binding-Site 110
[UniProt10a]
UniProt: Magnesium 1;
Metal-Binding-Site 112
[UniProt10a]
UniProt: Magnesium 1; via carbonyl oxygen;
Metal-Binding-Site 113
[UniProt10a]
UniProt: Magnesium 2;
Protein-Segment 113 -> 116
[UniProt10]
UniProt: Substrate binding; Sequence Annotation Type: region of interest;
Amino-Acid-Sites-That-Bind 187
[UniProt10a]
UniProt: Allosteric activator; via amide nitrogen;
Amino-Acid-Sites-That-Bind 206
[UniProt10a]
UniProt: Substrate;
Amino-Acid-Sites-That-Bind 222
[UniProt10a]
UniProt: Allosteric inhibitor glucose-6-phosphate;
Amino-Acid-Sites-That-Bind 225
[UniProt10a]
UniProt: Allosteric inhibitor glucose-6-phosphate;
Amino-Acid-Sites-That-Bind 239
[UniProt10a]
UniProt: Substrate;
Protein-Segment 257 -> 259
[UniProt10]
UniProt: Substrate binding; Sequence Annotation Type: region of interest;
Amino-Acid-Sites-That-Bind 269
[UniProt10a]
UniProt: Substrate;
Metal-Binding-Site 275
[UniProt10a]
UniProt: Magnesium 2;


Gene Local Context (not to scale): ?

Transcription Unit:

Notes:

History:
10/20/97 Gene b4232 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10283; confirmed by SwissProt match.


References

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

Babul83: Babul J, Guixe V (1983). "Fructose bisphosphatase from Escherichia coli. Purification and characterization." Arch Biochem Biophys 1983;225(2);944-9. PMID: 6312898

BRENDA14: BRENDA team (2014). "Imported from BRENDA version existing on Aug 2014." http://www.brenda-enzymes.org.

Brown09: Brown G, Singer A, Lunin VV, Proudfoot M, Skarina T, Flick R, Kochinyan S, Sanishvili R, Joachimiak A, Edwards AM, Savchenko A, Yakunin AF (2009). "Structural and biochemical characterization of the type II fructose-1,6-bisphosphatase GlpX from Escherichia coli." J Biol Chem 284(6);3784-92. PMID: 19073594

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

Fraenkel65: Fraenkel DG, Horecker BL (1965). "Fructose-1, 6-diphosphatase and acid hexose phosphatase of Escherichia coli." J Bacteriol 90(4);837-42. PMID: 4284917

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

GOA01: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

GOA01a: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

Hines06: Hines JK, Fromm HJ, Honzatko RB (2006). "Novel allosteric activation site in Escherichia coli fructose-1,6-bisphosphatase." J Biol Chem 281(27);18386-93. PMID: 16670087

Ishihama08: Ishihama Y, Schmidt T, Rappsilber J, Mann M, Hartl FU, Kerner MJ, Frishman D (2008). "Protein abundance profiling of the Escherichia coli cytosol." BMC Genomics 9;102. PMID: 18304323

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

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

Sedivy86: Sedivy JM, Babul J, Fraenkel DG (1986). "AMP-insensitive fructose bisphosphatase in Escherichia coli and its consequences." Proc Natl Acad Sci U S A 83(6);1656-9. PMID: 3006063

UniProt10: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-11 released on 2010-11-02 00:00:00." Database.

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

UniProtGOA12: UniProt-GOA (2012). "Gene Ontology annotation based on UniPathway vocabulary mapping."


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Sat Dec 20, 2014, BIOCYC13B.