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Escherichia coli K-12 substr. MG1655 Enzyme: phosphate acetyltransferase / phosphate propionyltransferase



Gene: pta Accession Numbers: EG20173 (EcoCyc), b2297, ECK2291

Regulation Summary Diagram: ?

Subunit composition of phosphate acetyltransferase / phosphate propionyltransferase = [Pta]6

Summary:
Phosphate acetyltransferase (Pta) catalyzes the reversible conversion between acetyl-CoA and acetylphosphate, a step in the metabolism of acetate. Both pyruvate and phosphoenolpyruvate activate the enzyme in the direction of acetylphosphate synthesis and inhibit the enzyme in the direction of acetyl-CoA synthesis [CamposBermudez10]. The acetate formation from acetyl-CoA I pathway has been the target of metabolic engineering to reduce the flux to acetate and increase the production of commercially desired end products; see, for example, [Dittrich05, De07, Shams08, Singh11a]. It has also been studied using systems biology approaches such as metabolic modeling and flux balance analysis; see, for example, [Fong06, Young08, Valgepea10].

Pta is composed of three domains; only the C-terminal domain is required for phosphate acetyltransferase activity. The N-terminal domain is involved in stabilization of the native quarternary structure and metabolic regulation [CamposBermudez10].

Pta may be able to utilize both acetyl-CoA and propionyl-CoA. An ack pta double mutant has reduced levels of propionate from L-threonine, suggesting that the enzyme is part of the anaerobic pathway metabolizing L-threonine to propionate [Hesslinger98] (see threonine degradation I).

A pta mutant does not grow on acetate as the sole source of carbon [Brown77]. Both pta and pta ackA mutants are impaired in their ability to survive glucose starvation [Nystrom94]. The growth defect of a pta mutant appears to be due to perturbation of acetyl-CoA flux [Chang99]. pta mutants produce large amounts of lactate when grown on glucose as the carbon source under microaerophilic conditions [Zhu05]. The effect of a pta mutation on metabolism, enzyme activity and gene expression has been thoroughly studied recently [CastanoCerezo09]. pta and recBC mutants are synthetically growth inhibited [Shi05a].

Levels of Pta are decreased by growth on acetate [Kirkpatrick01] and under low pH conditions [Stancik02]. pta belongs to the CreBC regulon [Avison01]. FNR has a slightly positive effect on pta expression [ShalelLevanon05]. The growth-rate dependent expression pattern of pta-ackA was measured [Nahku10].

Pta: "phosphotransacetylase" [Brown77]

Reviews: [ElMansi06, Wolfe05]

Citations: [Gupta89, Yang99a, Pascal81]

Gene Citations: [Kakuda94]

Locations: cytosol

Map Position: [2,412,769 -> 2,414,913] (52.0 centisomes)
Length: 2145 bp / 714 aa

Molecular Weight of Polypeptide: 77.172 kD (from nucleotide sequence), 81.0 kD (experimental) [YamamotoOtake90 ]

Molecular Weight of Multimer: 484.0 kD (experimental) [CamposBermudez10]

Unification Links: ASAP:ABE-0007582 , CGSC:353 , DIP:DIP-35815N , EchoBASE:EB4147 , EcoGene:EG20173 , EcoliWiki:b2297 , Mint:MINT-1263208 , ModBase:P0A9M8 , OU-Microarray:b2297 , PortEco:pta , PR:PRO_000023627 , Pride:P0A9M8 , Protein Model Portal:P0A9M8 , RefSeq:NP_416800 , RegulonDB:EG20173 , SMR:P0A9M8 , String:511145.b2297 , UniProt:P0A9M8

Relationship Links: InterPro:IN-FAMILY:IPR002505 , InterPro:IN-FAMILY:IPR004614 , InterPro:IN-FAMILY:IPR010766 , InterPro:IN-FAMILY:IPR016475 , InterPro:IN-FAMILY:IPR027417 , Pfam:IN-FAMILY:PF01515 , Pfam:IN-FAMILY:PF07085

In Paralogous Gene Group: 297 (4 members)

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0006083 - acetate metabolic process Inferred from experiment [Brown77]
GO:0019413 - acetate biosynthetic process Inferred from experiment [Brown77]
GO:0019427 - acetyl-CoA biosynthetic process from acetate Inferred from experiment [Brown77]
GO:0045733 - acetate catabolic process Inferred from experiment [Brown77]
GO:0070689 - L-threonine catabolic process to propionate Inferred from experiment [Hesslinger98]
GO:0006085 - acetyl-CoA biosynthetic process Inferred by computational analysis [UniProtGOA12]
GO:0008152 - metabolic process Inferred by computational analysis [GOA01]
Molecular Function: GO:0005515 - protein binding Inferred from experiment [Zheng11, Butland05]
GO:0008270 - zinc ion binding Inferred from experiment [Katayama02]
GO:0008959 - phosphate acetyltransferase activity Inferred from experiment Inferred by computational analysis [GOA01a, GOA01, CamposBermudez10, Goldman58]
GO:0016407 - acetyltransferase activity Inferred by computational analysis [GOA01]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11]
GO:0016746 - transferase activity, transferring acyl groups Inferred by computational analysis [UniProtGOA11, GOA01]
Cellular Component: GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [DiazMejia09, Ishihama08, LopezCampistrou05]
GO:0005737 - cytoplasm Inferred by computational analysis [UniProtGOA11a, UniProtGOA11]

MultiFun Terms: metabolism carbon utilization amino acids
metabolism carbon utilization carbon compounds
metabolism central intermediary metabolism acetate catabolism
metabolism central intermediary metabolism pyruvate oxidation
metabolism central intermediary metabolism threonine catabolism

Essentiality data for pta 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:
Curated 03-Dec-2010 by Keseler I , SRI International
Last-Curated ? 09-Nov-2011 by Fulcher C , SRI International


Enzymatic reaction of: phosphate acetyltransferase

Synonyms: phosphotransacetylase, phosphoacylase, acetyl-CoA:orthophosphate acetyltransferase

EC Number: 2.3.1.8

acetyl-CoA + phosphate <=> acetyl phosphate + coenzyme A

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

This reaction is reversible. [CamposBermudez10]

In Pathways: superpathway of acetate utilization and formation , acetate formation from acetyl-CoA I , mixed acid fermentation

Summary:
Purification and extensive characterization of the enzyme was reported from E. coli B [Suzuki69, Shimizu69, Suzuki69a].

The enzyme from E. coli K-12 has only recently been purified and characterized. The enzyme has an eight-fold lower kcat value for the acetylphosphate forming reaction. Km values for CoA and acetyl-CoA are S0.5 values rather than true Km values due to sigmoidal kinetics. Both pyruvate and phosphoenolpyruvate activate the enzyme in the direction of acetylphosphate synthesis and inhibit the enzyme in the direction of acetyl-CoA synthesis. The inhibitory effect of ATP and NADH is reversed in the presence of pyruvate or PEP [CamposBermudez10].

Activators (Unknown Mechanism): phosphoenolpyruvate [CamposBermudez10] , pyruvate [Suzuki69, CamposBermudez10]

Inhibitors (Noncompetitive): ATP [Suzuki69, CamposBermudez10, Comment 5] , ADP [Suzuki69, Comment 5] , NADPH [Suzuki69, Comment 5] , NADH [Suzuki69, CamposBermudez10, Comment 5]

Inhibitors (Unknown Mechanism): phosphoenolpyruvate [CamposBermudez10] , pyruvate [CamposBermudez10]

Kinetic Parameters:

Substrate
Km (μM)
Citations
phosphate
2100.0
[CamposBermudez10]
acetyl-CoA
44.9
[CamposBermudez10]
coenzyme A
67.2
[CamposBermudez10]
acetyl phosphate
900.0
[CamposBermudez10]


Enzymatic reaction of: phosphate propionyltransferase

EC Number: 2.3.1.222

propanoyl-CoA + phosphate <=> propanoyl phosphate + coenzyme A

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the Enzyme Commission system.

The reaction is favored in the direction shown.

In Pathways: superpathway of threonine metabolism , threonine degradation I


Sequence Features

Feature Class Location Citations Comment
Cleavage-of-Initial-Methionine 1
[Matsuyama94, Kakuda94, UniProt11]
UniProt: Removed.
Chain 2 -> 714
[UniProt09]
UniProt: Phosphate acetyltransferase;
Sequence-Conflict 20 -> 50
[Matsuyama94, UniProt10a]
Alternate sequence: SLGVIRAMERKGVRLSVFKPIAQPRTGGDAP → AWRDPCNGTQRRSSERFQTYRSAAYRWRCA; UniProt: (in Ref. 2; BAA04663);
Sequence-Conflict 208 -> 209
[Matsuyama94, Kakuda94, UniProt10a]
Alternate sequence: KL → NV; UniProt: (in Ref. 1 and 2);
Sequence-Conflict 263 -> 271
[Matsuyama94, UniProt10a]
Alternate sequence: SIPHMLEHF → QHSAHAGAL; UniProt: (in Ref. 2; BAA04663);
Protein-Segment 391 -> 714
[UniProt10]
UniProt: Phosphate acetyltransferase; Sequence Annotation Type: region of interest;


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
10/20/97 Gene b2297 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG20173; confirmed by SwissProt match.


References

Avison01: Avison MB, Horton RE, Walsh TR, Bennett PM (2001). "Escherichia coli CreBC is a global regulator of gene expression that responds to growth in minimal media." J Biol Chem 276(29);26955-61. PMID: 11350954

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

Brown77: Brown TD, Jones-Mortimer MC, Kornberg HL (1977). "The enzymic interconversion of acetate and acetyl-coenzyme A in Escherichia coli." J Gen Microbiol 1977;102(2);327-36. PMID: 21941

Butland05: Butland G, Peregrin-Alvarez JM, Li J, Yang W, Yang X, Canadien V, Starostine A, Richards D, Beattie B, Krogan N, Davey M, Parkinson J, Greenblatt J, Emili A (2005). "Interaction network containing conserved and essential protein complexes in Escherichia coli." Nature 433(7025);531-7. PMID: 15690043

CamposBermudez10: Campos-Bermudez VA, Bologna FP, Andreo CS, Drincovich MF (2010). "Functional dissection of Escherichia coli phosphotransacetylase structural domains and analysis of key compounds involved in activity regulation." FEBS J 277(8);1957-66. PMID: 20236319

CastanoCerezo09: Castano-Cerezo S, Pastor JM, Renilla S, Bernal V, Iborra JL, Canovas M (2009). "An insight into the role of phosphotransacetylase (pta) and the acetate/acetyl-CoA node in Escherichia coli." Microb Cell Fact 8;54. PMID: 19852855

Chang99: Chang DE, Shin S, Rhee JS, Pan JG (1999). "Acetate metabolism in a pta mutant of Escherichia coli W3110: importance of maintaining acetyl coenzyme A flux for growth and survival." J Bacteriol 181(21);6656-63. PMID: 10542166

De07: De Mey M, Lequeux GJ, Beauprez JJ, Maertens J, Van Horen E, Soetaert WK, Vanrolleghem PA, Vandamme EJ (2007). "Comparison of different strategies to reduce acetate formation in Escherichia coli." Biotechnol Prog 23(5);1053-63. PMID: 17715942

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

Dittrich05: Dittrich CR, Bennett GN, San KY (2005). "Characterization of the acetate-producing pathways in Escherichia coli." Biotechnol Prog 21(4);1062-7. PMID: 16080684

ElMansi06: El-Mansi M, Cozzone AJ, Shiloach J, Eikmanns BJ (2006). "Control of carbon flux through enzymes of central and intermediary metabolism during growth of Escherichia coli on acetate." Curr Opin Microbiol 9(2);173-9. PMID: 16530464

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

Fong06: Fong SS, Nanchen A, Palsson BO, Sauer U (2006). "Latent pathway activation and increased pathway capacity enable Escherichia coli adaptation to loss of key metabolic enzymes." J Biol Chem 281(12);8024-33. PMID: 16319065

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

GOA01a: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

Goldman58: Goldman DS (1958). "Purification of phosphotransacetylase from Escherichia coli, K-12." Biochim Biophys Acta 28(2);436-7. PMID: 13535743

Gupta89: Gupta S, Clark DP (1989). "Escherichia coli derivatives lacking both alcohol dehydrogenase and phosphotransacetylase grow anaerobically by lactate fermentation." J Bacteriol 171(7);3650-5. PMID: 2661531

Hesslinger98: Hesslinger C, Fairhurst SA, Sawers G (1998). "Novel keto acid formate-lyase and propionate kinase enzymes are components of an anaerobic pathway in Escherichia coli that degrades L-threonine to propionate." Mol Microbiol 1998;27(2);477-92. PMID: 9484901

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

Kakuda94: Kakuda H, Hosono K, Shiroishi K, Ichihara S (1994). "Identification and characterization of the ackA (acetate kinase A)-pta (phosphotransacetylase) operon and complementation analysis of acetate utilization by an ackA-pta deletion mutant of Escherichia coli." J Biochem 116(4);916-22. PMID: 7883769

Katayama02: Katayama A, Tsujii A, Wada A, Nishino T, Ishihama A (2002). "Systematic search for zinc-binding proteins in Escherichia coli." Eur J Biochem 269(9);2403-13. PMID: 11985624

Kirkpatrick01: Kirkpatrick C, Maurer LM, Oyelakin NE, Yoncheva YN, Maurer R, Slonczewski JL (2001). "Acetate and formate stress: opposite responses in the proteome of Escherichia coli." J Bacteriol 183(21);6466-77. PMID: 11591692

LeVine80: LeVine SM, Ardeshir F, Ames GF (1980). "Isolation and Characterization of acetate kinase and phosphotransacetylase mutants of Escherichia coli and Salmonella typhimurium." J Bacteriol 143(2);1081-5. PMID: 6259116

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

Matsuyama94: Matsuyama A, Yamamoto-Otake H, Hewitt J, MacGillivray RT, Nakano E (1994). "Nucleotide sequence of the phosphotransacetylase gene of Escherichia coli strain K12." Biochim Biophys Acta 1219(2);559-62. PMID: 7918659

Nahku10: Nahku R, Valgepea K, Lahtvee PJ, Erm S, Abner K, Adamberg K, Vilu R (2010). "Specific growth rate dependent transcriptome profiling of Escherichia coli K12 MG1655 in accelerostat cultures." J Biotechnol 145(1);60-5. PMID: 19861135

Nystrom94: Nystrom T (1994). "The glucose-starvation stimulon of Escherichia coli: induced and repressed synthesis of enzymes of central metabolic pathways and role of acetyl phosphate in gene expression and starvation survival." Mol Microbiol 12(5);833-43. PMID: 8052134

Pascal81: Pascal MC, Chippaux M, Abou-Jaoude A, Blaschkowski HP, Knappe J (1981). "Mutants of Escherichia coli K12 with defects in anaerobic pyruvate metabolism." J Gen Microbiol 1981;124(Pt 1);35-42. PMID: 7033467

ShalelLevanon05: Shalel-Levanon S, San KY, Bennett GN (2005). "Effect of ArcA and FNR on the expression of genes related to the oxygen regulation and the glycolysis pathway in Escherichia coli under microaerobic growth conditions." Biotechnol Bioeng 92(2):147-59. PMID: 15988767

Shams08: Shams Yazdani S, Gonzalez R (2008). "Engineering Escherichia coli for the efficient conversion of glycerol to ethanol and co-products." Metab Eng 10(6);340-51. PMID: 18840539

Shi05a: Shi IY, Stansbury J, Kuzminov A (2005). "A defect in the acetyl coenzyme A<-->acetate pathway poisons recombinational repair-deficient mutants of Escherichia coli." J Bacteriol 187(4);1266-75. PMID: 15687190

Shimizu69: Shimizu M, Suzuki T, Kameda KY, Abiko Y (1969). "Phosphotransacetylase of Escherichia coli B, purification and properties." Biochim Biophys Acta 191(3);550-8. PMID: 4903502

Singh11a: Singh A, Cher Soh K, Hatzimanikatis V, Gill RT (2011). "Manipulating redox and ATP balancing for improved production of succinate in E. coli." Metab Eng 13(1);76-81. PMID: 21040799

Stancik02: Stancik LM, Stancik DM, Schmidt B, Barnhart DM, Yoncheva YN, Slonczewski JL (2002). "pH-dependent expression of periplasmic proteins and amino acid catabolism in Escherichia coli." J Bacteriol 184(15);4246-58. PMID: 12107143

Suzuki69: Suzuki T (1969). "Phosphotransacetylase of Escherichia coli B, activation by pyruvate and inhibition by NADH and certain nucleotides." Biochim Biophys Acta 1969;191(3);559-69. PMID: 4312205

Suzuki69a: Suzuki T, Abiko Y, Shimizu M (1969). "Activation and inhibition of purified phosphotransacetylase of Escherichia coli B by pyruvate and by NADH2 and certain nucleotides." Biochem Biophys Res Commun 35(1);102-8. PMID: 4305270

UniProt09: UniProt Consortium (2009). "UniProt version 15.8 released on 2009-10-01 00:00:00." Database.

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.

UniProt11: UniProt Consortium (2011). "UniProt version 2011-06 released on 2011-06-30 00:00:00." Database.

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

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

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

Valgepea10: Valgepea K, Adamberg K, Nahku R, Lahtvee PJ, Arike L, Vilu R (2010). "Systems biology approach reveals that overflow metabolism of acetate in Escherichia coli is triggered by carbon catabolite repression of acetyl-CoA synthetase." BMC Syst Biol 4(1);166. PMID: 21122111

Wolfe05: Wolfe AJ (2005). "The acetate switch." Microbiol Mol Biol Rev 69(1);12-50. PMID: 15755952

YamamotoOtake90: Yamamoto-Otake H, Matsuyama A, Nakano E (1990). "Cloning of a gene coding for phosphotransacetylase from Escherichia coli." Appl Microbiol Biotechnol 33(6);680-2. PMID: 1366944

Yang99a: Yang YT, Bennett GN, San KY (1999). "Effect of inactivation of nuo and ackA-pta on redistribution of metabolic fluxes in Escherichia coli." Biotechnol Bioeng 65(3);291-7. PMID: 10486127

Young08: Young JD, Henne KL, Morgan JA, Konopka AE, Ramkrishna D (2008). "Integrating cybernetic modeling with pathway analysis provides a dynamic, systems-level description of metabolic control." Biotechnol Bioeng 100(3);542-59. PMID: 18438875

Zheng11: Zheng C, Yang L, Hoopmann MR, Eng JK, Tang X, Weisbrod CR, Bruce JE (2011). "Cross-linking measurements of in vivo protein complex topologies." Mol Cell Proteomics 10(10);M110.006841. PMID: 21697552

Zhu05: Zhu J, Shimizu K (2005). "Effect of a single-gene knockout on the metabolic regulation in Escherichia coli for D-lactate production under microaerobic condition." Metab Eng 7(2);104-15. PMID: 15781419

Other References Related to Gene Regulation

Marzan13: Marzan LW, Hasan CM, Shimizu K (2013). "Effect of acidic condition on the metabolic regulation of Escherichia coli and its phoB mutant." Arch Microbiol 195(3);161-71. PMID: 23274360


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
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