Escherichia coli K-12 substr. MG1655 Enzyme: manno(fructo)kinase

Gene: mak Accession Numbers: EG11288 (EcoCyc), b0394, ECK0389

Synonyms: yajF

Regulation Summary Diagram: ?

Regulation summary diagram for mak

In the absence of the usual PTS-mediated route of fructose uptake, Mak catalyzes phosphorylation of fructose after its entry into the cell via PtsGF [Kornberg00].

Mak shows trace activity in wild-type cells [Kornberg01]. An A24D mutation causes increased Mak activity [Kornberg01]. The A24D mutant protein is more stable than wild-type Mak [Sproul01].

Overexpression of yajF rescues the glucose auxotrophy of a glucokinase mutant, and the YajF protein functions as a rudimentary glucokinase in vitro [Miller04]. When a glucokinase-deficient strain is placed under selective pressure for growth on glucose, a spontaneous mutation in the predicted promoter region of yajF arises, regenerating a sigma70 consensus sequence and leading to a 94-fold increase in the expression of yajF [Miller05].

Review: [Kornberg01].
The start site of the mak gene was originally assigned just based on sequence considerations [Blattner97]. However, it was changed because [Sproul01] showed that the real start site is actually located 138 bp downstream and its ATG start codon was re-annotated at the 409368 position. The demonstration is based on sequence analysis, and the identification of both a plausible ribosome binding site and a promoter at an appropriate distance from the new start site [Sproul01, Miller05, Miller04]. The promoter was identified using mutational analysis [Miller05]. This information was obtained from the EcoGene data base (

Locations: cytosol

Map Position: [409,368 -> 410,276] (8.82 centisomes, 32°)
Length: 909 bp / 302 aa

Molecular Weight of Polypeptide: 32.5 kD (from nucleotide sequence)

Unification Links: ASAP:ABE-0001372 , DIP:DIP-11288N , EchoBASE:EB1265 , EcoGene:EG11288 , EcoliWiki:b0394 , ModBase:P23917 , OU-Microarray:b0394 , PortEco:mak , PR:PRO_000023146 , Pride:P23917 , Protein Model Portal:P23917 , RefSeq:NP_414928 , RegulonDB:EG11288 , SMR:P23917 , String:511145.b0394 , UniProt:P23917

Relationship Links: InterPro:IN-FAMILY:IPR000600 , Pfam:IN-FAMILY:PF00480 , Prosite:IN-FAMILY:PS01125

In Paralogous Gene Group: 118 (7 members)

Gene-Reaction Schematic: ?

Gene-Reaction Schematic

GO Terms:

Biological Process: GO:0019318 - hexose metabolic process Inferred from experiment [Miller04]
GO:0046835 - carbohydrate phosphorylation Inferred from experiment Inferred by computational analysis [GOA01, Kornberg00]
GO:0005975 - carbohydrate metabolic process Inferred by computational analysis [UniProtGOA11a]
GO:0016310 - phosphorylation Inferred by computational analysis [UniProtGOA11a]
Molecular Function: GO:0004396 - hexokinase activity Inferred from experiment [Kornberg00]
GO:0000166 - nucleotide binding Inferred by computational analysis [UniProtGOA11a]
GO:0005524 - ATP binding Inferred by computational analysis [UniProtGOA11a]
GO:0008865 - fructokinase activity Inferred by computational analysis [GOA01]
GO:0016301 - kinase activity Inferred by computational analysis [UniProtGOA11a]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [DiazMejia09, Ishihama08, Kornberg00]

MultiFun Terms: metabolism carbon utilization carbon compounds

Essentiality data for mak 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]

Enzymatic reaction of: manno(fructo)kinase

Synonyms: fructokinase, ATP:D-fructose 6-phosphotransferase

EC Number:

β-D-fructofuranose + ATP <=> β-D-fructofuranose 6-phosphate + ADP + H+

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.

Sequence Features

Protein sequence of manno(fructo)kinase with features indicated

Feature Class Location Citations Comment
Nucleotide-Phosphate-Binding-Region 4 -> 11
UniProt: ATP; Non-Experimental Qualifier: potential;
Nucleotide-Phosphate-Binding-Region 133 -> 140
UniProt: ATP; Non-Experimental Qualifier: potential;

Gene Local Context (not to scale): ?

Gene local context diagram

Transcription Unit:

Transcription-unit diagram


10/20/97 Gene b0394 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG11288; 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

Blattner97: Blattner FR, Plunkett G, Bloch CA, Perna NT, Burland V, Riley M, Collado-Vides J, Glasner JD, Rode CK, Mayhew GF, Gregor J, Davis NW, Kirkpatrick HA, Goeden MA, Rose DJ, Mau B, Shao Y (1997). "The complete genome sequence of Escherichia coli K-12." Science 277(5331);1453-74. PMID: 9278503

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

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.

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

Kornberg00: Kornberg HL, Lambourne LT, Sproul AA (2000). "Facilitated diffusion of fructose via the phosphoenolpyruvate/glucose phosphotransferase system of Escherichia coli." Proc Natl Acad Sci U S A 97(4);1808-12. PMID: 10677538

Kornberg01: Kornberg HL (2001). "Routes for fructose utilization by Escherichia coli." J Mol Microbiol Biotechnol 3(3);355-9. PMID: 11361065

Miller04: Miller BG, Raines RT (2004). "Identifying latent enzyme activities: substrate ambiguity within modern bacterial sugar kinases." Biochemistry 43(21);6387-92. PMID: 15157072

Miller05: Miller BG, Raines RT (2005). "Reconstitution of a defunct glycolytic pathway via recruitment of ambiguous sugar kinases." Biochemistry 44(32);10776-83. PMID: 16086580

Sproul01: Sproul AA, Lambourne LT, Jean-Jacques D J, Kornberg HL (2001). "Genetic control of manno(fructo)kinase activity in Escherichia coli." Proc Natl Acad Sci U S A 98(26);15257-9. PMID: 11742072

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

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

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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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