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Escherichia coli K-12 substr. MG1655 Enzyme: S-formylglutathione hydrolase / S-lactoylglutathione hydrolase



Gene: yeiG Accession Numbers: EG12026 (EcoCyc), b2154, ECK2147

Regulation Summary Diagram: ?

Regulation summary diagram for yeiG

Subunit composition of S-formylglutathione hydrolase / S-lactoylglutathione hydrolase = [YeiG]4

Summary:
YeiG is a promiscuous serine hydrolase; its highest specific activity is with the substrate S-formylglutathione. Sulfhydryl inhibitors affect enzymatic activity [Gonzalez06]. A general esterase activity of YeiG was first discovered in a high-throughput screen of purified proteins [Kuznetsova05].

YeiG also has significant activity with the substrate lactoylglutathione, an intermediate of the detoxification of methylglyoxal. It may thus represent a cytoplasmic equivalent of glyoxalase II, which may be involved in the detoxification of endogenous methylglyoxal [Gonzalez06].

YeiG has similarity to S-formylglutathione hydrolases of Arabidopsis, S. cerevisiae, human, and a paralog, FrmB [Gonzalez06].

yeiG is expressed constitutively in both the wild type and frmB deletion background. Under normal growth conditions, neither a yeiG deletion mutant nor a frmB yeiG double mutant have a detectable growth defect. Addition of 0.4 mM formaldehyde to the growth medium had no effect on the yeiG single mutant, while the growth rate of the double mutant drops to 43% of wild type [Gonzalez06]. Increased expression of yeiG is one of the contributing factors in an L-glutamine-overproducing mutant [Hayashi13]. Site-directed yeiG mutants were used to define residues important for catalytic activity [Gonzalez06].

Map Position: [2,241,932 -> 2,242,768] (48.32 centisomes, 174°)
Length: 837 bp / 278 aa

Molecular Weight of Polypeptide: 31.259 kD (from nucleotide sequence)

Molecular Weight of Multimer: 121.2 kD (experimental) [Gonzalez06]

Unification Links: ASAP:ABE-0007124 , DIP:DIP-11919N , EchoBASE:EB1961 , EcoGene:EG12026 , EcoliWiki:b2154 , ModBase:P33018 , OU-Microarray:b2154 , PortEco:yeiG , Protein Model Portal:P33018 , RefSeq:NP_416659 , RegulonDB:EG12026 , SMR:P33018 , String:511145.b2154 , UniProt:P33018

Relationship Links: InterPro:IN-FAMILY:IPR000801 , InterPro:IN-FAMILY:IPR014186 , InterPro:IN-FAMILY:IPR029058 , Panther:IN-FAMILY:PTHR10061 , Pfam:IN-FAMILY:PF00756

Gene-Reaction Schematic: ?

Gene-Reaction Schematic

GO Terms:

Biological Process: GO:0046294 - formaldehyde catabolic process Inferred from experiment Inferred by computational analysis [GOA01, Gonzalez06]
GO:0051289 - protein homotetramerization Inferred from experiment [Gonzalez06]
GO:0019243 - methylglyoxal catabolic process to D-lactate Author statement [Gonzalez06]
Molecular Function: GO:0004416 - hydroxyacylglutathione hydrolase activity Inferred from experiment [Gonzalez06]
GO:0018738 - S-formylglutathione hydrolase activity Inferred from experiment Inferred by computational analysis [GOA01a, GOA01, Gonzalez06]
GO:0042802 - identical protein binding Inferred from experiment [Gonzalez06]
GO:0016787 - hydrolase activity Inferred by computational analysis [UniProtGOA11]
GO:0052689 - carboxylic ester hydrolase activity Inferred by computational analysis [UniProtGOA11]

MultiFun Terms: cell processes protection detoxification

Essentiality data for yeiG 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:
Created 29-Mar-2006 by Keseler I , SRI International
Last-Curated ? 27-Mar-2015 by Keseler I , SRI International


Enzymatic reaction of: S-formylglutathione hydrolase

EC Number: 3.1.2.12

S-formylglutathione + H2O <=> formate + glutathione + H+

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.

Alternative Substrates for S-formylglutathione: α-naphthyl acetate [Gonzalez06 ]

In Pathways: formaldehyde oxidation II (glutathione-dependent)

Summary:
The enzyme shows a sigmoidal saturation curve, indicating positive cooperativity in S-formylglutathione binding. High concentrations of S-formylglutathione inhibit the enzymatic activity [Gonzalez06].

Inhibitors (Unknown Mechanism): Cu2+ [Gonzalez06] , Zn2+ [Gonzalez06] , N-ethylmaleimide [Gonzalez06] , iodoacetate [Gonzalez06]

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
S-formylglutathione
430.0
6.51
0.015
[Gonzalez06]


Enzymatic reaction of: S-lactoylglutathione hydrolase

EC Number: 3.1.2.6

(R)-S-lactoylglutathione + H2O <=> glutathione + (R)-lactate + H+

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 methylglyoxal degradation , methylglyoxal degradation I

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
(R)-S-lactoylglutathione
580.0
1.02
0.0018
[Gonzalez06]


Sequence Features

Protein sequence of YeiG with features indicated

Feature Class Location Citations Comment
Mutagenesis-Variant 26
[Gonzalez06, UniProt11]
UniProt: Reduces catalytic efficiency, but has no effect on affinity for substrate.
Mutagenesis-Variant 54
[Gonzalez06, UniProt11]
UniProt: Reduces affinity for substrate and catalytic efficiency.
Mutagenesis-Variant 80
[Gonzalez06, UniProt11]
UniProt: Increases affinity for substrate, but reduces catalytic efficiency.
Mutagenesis-Variant 145
[Gonzalez06, UniProt11]
UniProt: Loss of activity.
Active-Site 145
[UniProt10]
UniProt: Charge relay system; Non-Experimental Qualifier: probable;
Mutagenesis-Variant 199
[Gonzalez06, UniProt11]
UniProt: Reduces catalytic efficiency, but has no effect on affinity for substrate.
Mutagenesis-Variant 218
[Gonzalez06, UniProt11]
UniProt: Reduces affinity for substrate and catalytic efficiency.
Mutagenesis-Variant 223
[Gonzalez06, UniProt11]
UniProt: Loss of activity.
Active-Site 223
[UniProt10]
UniProt: Charge relay system; Non-Experimental Qualifier: probable;
Mutagenesis-Variant 255
[Gonzalez06, UniProt11]
UniProt: Increases affinity for substrate, but reduces catalytic efficiency.
Mutagenesis-Variant 256
[Gonzalez06, UniProt11]
UniProt: Loss of activity.
Active-Site 256
[UniProt10]
UniProt: Charge relay system; Non-Experimental Qualifier: probable;


Gene Local Context (not to scale): ?

Gene local context diagram

Transcription Unit:

Transcription-unit diagram

Notes:

History:
10/20/97 Gene b2154 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG12026; 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

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, 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.

Gonzalez06: Gonzalez CF, Proudfoot M, Brown G, Korniyenko Y, Mori H, Savchenko AV, Yakunin AF (2006). "Molecular basis of formaldehyde detoxification: Characterization of two s-formylglutathione hydrolases from Escherichia coli, FrmB and YeiG." J Biol Chem 281:14514-14522. PMID: 16567800

Hayashi13: Hayashi M, Tabata K (2013). "Metabolic engineering for L-glutamine overproduction by using DNA gyrase mutations in Escherichia coli." Appl Environ Microbiol 79(9);3033-9. PMID: 23455340

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

Kuznetsova05: Kuznetsova E, Proudfoot M, Sanders SA, Reinking J, Savchenko A, Arrowsmith CH, Edwards AM, Yakunin AF (2005). "Enzyme genomics: Application of general enzymatic screens to discover new enzymes." FEMS Microbiol Rev 29(2);263-79. PMID: 15808744

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


Report Errors or Provide Feedback
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 19.0 on Wed Sep 2, 2015, biocyc14.