Gene Symbol: gshA
Description: glutamate-cysteine ligase
Alias: ECK2683, JW2663, gshI
Species: Escherichia coli str. K-12 substr. MG1655
Products:     gshA

Top Publications

  1. Li H, Li Y, Lin J, Chen J. [Construction of recombinant E. coli with high glutathione biosynthetic activity and the biosynthetic process]. Wei Sheng Wu Xue Bao. 2001;41:16-24 pubmed
    ..Further degradation of GSH was prevented by the addition of 100 mmol/L L-serine and potassium borate mixture. In such case, 23.0 mmol/L (about 7.1 g/L) GSH accumulated at 3 h. ..
  2. Kelly B, Antholine W, Griffith O. Escherichia coli gamma-glutamylcysteine synthetase. Two active site metal ions affect substrate and inhibitor binding. J Biol Chem. 2002;277:50-8 pubmed
    ..The results are discussed in the context of the catalytic mechanism of gamma-GCS and its relationship to the more fully characterized glutamine synthetase reaction. ..
  3. Benov L, Fridovich I. Escherichia coli exhibits negative chemotaxis in gradients of hydrogen peroxide, hypochlorite, and N-chlorotaurine: products of the respiratory burst of phagocytic cells. Proc Natl Acad Sci U S A. 1996;93:4999-5002 pubmed
    ..Hypochlorite and N-chlorotaurine were also strongly repellent. Chemotaxis down gradients of H2O2, OCl-, and N-chlorotaurine may contribute to the survival of commensal or pathogenic microorganisms. ..
  4. Gallardo Madueño R, Leal J, Dorado G, Holmgren A, Lopez Barea J, Pueyo C. In vivo transcription of nrdAB operon and of grxA and fpg genes is triggered in Escherichia coli lacking both thioredoxin and glutaredoxin 1 or thioredoxin and glutathione, respectively. J Biol Chem. 1998;273:18382-8 pubmed
    ..The increased expression of the grxA gene in trxA gshA double mutant bacteria was mimicked in trxA single mutant cells by depletion of GSH with diethylmaleate (DEM)...
  5. Waldman H. Baby fat is cute, but chubby kids may be in danger. ASDC J Dent Child. 2000;67:15-7, 8 pubmed
    ..An increasing percent of children are overweight. A review of the prevalence of overweight children in different demographic groups is provided in an effort to alert dentists to the role they could play in preventing medical complications. ..
  6. Hiratake J, Irie T, Tokutake N, Oda J. Recognition of a cysteine substrate by E. coli gamma-glutamylcysteine synthetase probed by sulfoximine-based transition-state analogue inhibitors. Biosci Biotechnol Biochem. 2002;66:1500-14 pubmed
  7. Freedberg W, Kistler W, Lin E. Lethal synthesis of methylglyoxal by Escherichia coli during unregulated glycerol metabolism. J Bacteriol. 1971;108:137-44 pubmed
    ..Methylglyoxal-resistant mutants derived from the glycerol-sensitive parental strain also became immune to glycerol. ..
  8. Gardner P, Fridovich I. Effect of glutathione on aconitase in Escherichia coli. Arch Biochem Biophys. 1993;301:98-102 pubmed
    ..A mutant deficient in GSH biosynthesis, designated gshA, grew slower in a defined medium than did the parental strain and this effect was more pronounced when succinate ..
  9. Hibi T, Nii H, Nakatsu T, Kimura A, Kato H, Hiratake J, et al. Crystal structure of gamma-glutamylcysteine synthetase: insights into the mechanism of catalysis by a key enzyme for glutathione homeostasis. Proc Natl Acad Sci U S A. 2004;101:15052-7 pubmed
    ..coli gammaGCS and its eukaryotic homologues. The identification of this cysteine-binding site provides important information for the rational design of novel gammaGCS inhibitors. ..

More Information


  1. Murata K, Kimura A. Some properties of glutathione biosynthesis-deficient mutants of Escherichia coli B. J Gen Microbiol. 1982;128:1047-52 pubmed
    ..The mutants lacking gamma-glutamylcysteine synthetase activity were also susceptible to oxygen. ..
  2. Oktyabrsky O, Smirnovam G, Muzyka N. Role of glutathione in regulation of hydroperoxidase I in growing Escherichia coli. Free Radic Biol Med. 2001;31:250-5 pubmed
    ..In the absence of H2O2 and in the presence of the low H2O2 concentrations (0.1-3 mM), the gshA mutation stimulated katG::lacZ expression and the total catalase activity in exponential phase...
  3. Hibi T, Hisada H, Nakatsu T, Kato H, Oda J. Escherichia coli B gamma-glutamylcysteine synthetase: modification, purification, crystallization and preliminary crystallographic analysis. Acta Crystallogr D Biol Crystallogr. 2002;58:316-8 pubmed
    ..The gammaGCS crystal diffracted to 2.8 A and belongs to space group R3, with unit-cell parameters a = b = 326.7, c = 103.9 A. ..
  4. Pacheco C, Passos J, Castro A, Moradas Ferreira P, De Marco P. Role of respiration and glutathione in cadmium-induced oxidative stress in Escherichia coli K-12. Arch Microbiol. 2008;189:271-8 pubmed
  5. Huang C, Moore W, Meister A. On the active site thiol of gamma-glutamylcysteine synthetase: relationships to catalysis, inhibition, and regulation. Proc Natl Acad Sci U S A. 1988;85:2464-8 pubmed
    ..If the active site thiol of the rat kidney enzyme is involved in catalysis, which seems likely, there would appear to be differences in the mechanisms of action of the two gamma-glutamylcysteine synthetases. ..
  6. Smirnova G, Krasnykh T, Oktyabrsky O. Role of glutathione in the response of Escherichia coli to osmotic stress. Biochemistry (Mosc). 2001;66:973-8 pubmed
    The growth of Escherichia coli mutants deficient in glutathione synthesis (gshA) and in glutathione reductase (gor) was suppressed in medium of elevated osmolarity...
  7. Murata K, Kimura A. Cloning of a gene responsible for the biosynthesis of glutathione in Escherichia coli B. Appl Environ Microbiol. 1982;44:1444-8 pubmed
    A gene (gshI) responsible for gamma-glutamylcysteine synthetase (GSH-I) activity was cloned to construct an Escherichia coli B strain having high glutathione synthesizing activity. For this purpose, two E...
  8. Oden K, Gladysheva T, Rosen B. Arsenate reduction mediated by the plasmid-encoded ArsC protein is coupled to glutathione. Mol Microbiol. 1994;12:301-6 pubmed the ars genes, arsenate resistance required the products of the gor gene for glutathione reductase and the gshA and gshB genes for glutathione synthesis...
  9. Kumar S, Kasturia N, Sharma A, Datt M, Bachhawat A. Redox-dependent stability of the ?-glutamylcysteine synthetase enzyme of Escherichia coli: a novel means of redox regulation. Biochem J. 2013;449:783-94 pubmed publisher
    ..These results demonstrate a novel means of regulation of ?-GCS by the redox environment that works by an alteration in its stability. ..
  10. Pittman M, Robinson H, Poole R. A bacterial glutathione transporter (Escherichia coli CydDC) exports reductant to the periplasm. J Biol Chem. 2005;280:32254-61 pubmed
    ..swarming motility and benzylpenicillin sensitivity in a cydD mutant and also benzylpenicillin sensitivity in a gshA mutant defective in GSH synthesis...
  11. Sakamoto A, Terui Y, Yoshida T, Yamamoto T, Suzuki H, Yamamoto K, et al. Three members of polyamine modulon under oxidative stress conditions: two transcription factors (SoxR and EmrR) and a glutathione synthetic enzyme (GshA). PLoS ONE. 2015;10:e0124883 pubmed publisher
    ..a negative transcriptional factor for expression of the genes for drug excretion proteins, EmrA and EmrB, and of GshA, γ-glutamylcysteine synthetase necessary for glutathione (GSH) synthesis, were stimulated by polyamines at the ..
  12. Shen L, Wei D, Zhao Z, Zhang S, Wang E. [Cloning and expression of the genes of glutathione synthetases]. Sheng Wu Gong Cheng Xue Bao. 2001;17:98-100 pubmed
    ..7 after the addition of 0.1 mmol/L IPTG. The expressed products were up to 25% of the total protein of the bacteria. Acetone-treated cells of the engineered strain could synthesize GSH efficiently. ..
  13. Helbig K, Bleuel C, Krauss G, Nies D. Glutathione and transition-metal homeostasis in Escherichia coli. J Bacteriol. 2008;190:5431-8 pubmed publisher
    ..Thus, GSH may play an important role in trace-element metabolism not only in higher organisms but also in bacteria. ..
  14. Miyake K, Kakita S. A novel catalytic ability of gamma-glutamylcysteine synthetase of Escherichia coli and its application in theanine production. Biosci Biotechnol Biochem. 2009;73:2677-83 pubmed
    ..The resting cells of E. coli expressing gammaGCS, in which ATP was regenerated through glycolysis, synthesized 12.1 mM theanine (18 h) from 429 mM ethylamine. ..
  15. Greenberg J, Demple B. Glutathione in Escherichia coli is dispensable for resistance to H2O2 and gamma radiation. J Bacteriol. 1986;168:1026-9 pubmed
    Escherichia coli devoid of glutathione (because of transposon insertions in the gshA gene) has normal resistance to H2O2, cumene hydroperoxide, heat, or ionizing radiation. Intracellular glutathione thus does not protect E...
  16. Smirnova G, Muzyka N, Oktyabrsky O. Effects of cystine and hydrogen peroxide on glutathione status and expression of antioxidant genes in Escherichia coli. Biochemistry (Mosc). 2005;70:926-34 pubmed
    ..The high toxicity of H2O2 in the presence of cystine resulted in disorders of membrane functions and inhibition of the expression of genes including those responsible for neutralization of oxidants and DNA repair. ..
  17. Kato H, Tanaka T, Nishioka T, Kimura A, Oda J. Role of cysteine residues in glutathione synthetase from Escherichia coli B. Chemical modification and oligonucleotide site-directed mutagenesis. J Biol Chem. 1988;263:11646-51 pubmed
    ..Replacement of Cys-122 to Ala-122 enhanced the reactivity of Cys-289 with sulfhydryl reagents. ..
  18. Inoue Y, Iba Y, Yano H, Murata K, Kimura A. Functional analysis of the gamma-glutamylcysteine synthetase of Escherichia coli B: effect of substitution of His-150 to Ala. Appl Microbiol Biotechnol. 1993;38:473-7 pubmed
    ..Among them, His residues were substituted to Ala by site-directed mutagenesis, and His-150 was found to be essential for the activity of GSH-I. ..
  19. Tran Q, Arras T, Becker S, Holighaus G, Ohlberger G, Unden G. Role of glutathione in the formation of the active form of the oxygen sensor FNR ([4Fe-4S].FNR) and in the control of FNR function. Eur J Biochem. 2000;267:4817-24 pubmed
    ..In a gshA mutant of E...
  20. Toledano M, Kumar C, Le Moan N, Spector D, Tacnet F. The system biology of thiol redox system in Escherichia coli and yeast: differential functions in oxidative stress, iron metabolism and DNA synthesis. FEBS Lett. 2007;581:3598-607 pubmed
    ..We show that although prokaryotic and eukaryotic systems have a similar architecture, they profoundly differ in their overall cellular functions. ..
  21. Moore W, Anderson M, Meister A, Murata K, Kimura A. Increased capacity for glutathione synthesis enhances resistance to radiation in Escherichia coli: a possible model for mammalian cell protection. Proc Natl Acad Sci U S A. 1989;86:1461-4 pubmed
    ..Conclusions drawn from studies on this E. coli model system may have relevance to protection of mammalian cells by glutathione. ..
  22. Apontoweil P, Berends W. Isolation and initial characterization of glutathione-deficient mutants of Escherichia coli K 12. Biochim Biophys Acta. 1975;399:10-22 pubmed
    ..It must be concluded that the protective role of glutathione is only significant when a chemical challenge is present. ..
  23. Tokutake N, Hiratake J, Katoh M, Irie T, Kato H, Oda J. Design, synthesis and evaluation of transition-state analogue inhibitors of Escherichia coli gamma-glutamylcysteine synthetase. Bioorg Med Chem. 1998;6:1935-53 pubmed
    ..The sulfoximine with (S)-sulfur was a simple reversible inhibitor with an inhibition potency comparable to that of the sulfone. The synthesis and inhibition profile of the N-phosphoryl sulfoximine is also described. ..
  24. Sleigh M, Grigg G. Sulphydryl-mediated DNA breakage by phlemomycin in Escherichia coli. Mutat Res. 1977;42:181-90 pubmed
    ..In spheroplasts of these gshA mutants, restoration of the response to phleomycin of dithiothreitol...
  25. Miranda Vizuete A, Rodriguez Ariza A, Toribio F, Holmgren A, Lopez Barea J, Pueyo C. The levels of ribonucleotide reductase, thioredoxin, glutaredoxin 1, and GSH are balanced in Escherichia coli K12. J Biol Chem. 1996;271:19099-103 pubmed
    ..redox status in new Escherichia coli K12 strains lacking thioredoxin (trxA-), glutaredoxin 1 (grxA-), and/or GSH (gshA-) or overproducing Trx or Grx1 from multicopy plasmids...
  26. Harrison J, Tremaroli V, Stan M, Chan C, Vacchi Suzzi C, Heyne B, et al. Chromosomal antioxidant genes have metal ion-specific roles as determinants of bacterial metal tolerance. Environ Microbiol. 2009;11:2491-509 pubmed publisher
    ..This characterized approximately 250 gene-metal combinations and identified that sodA, sodB, gor, trxA, gshA, grxA and marR have distinct roles in safeguarding or sensitizing cells to different toxic metal ions (Cr(2)O(7)(2-)..
  27. Helbig K, Grosse C, Nies D. Cadmium toxicity in glutathione mutants of Escherichia coli. J Bacteriol. 2008;190:5439-54 pubmed publisher
    ..General energy conservation pathways and iron uptake were down-regulated. These findings indicated that the toxic action of Cd(2+) indeed results from the binding of the metal cation to sulfur, lending support to the hypothesis tested. ..
  28. Faulkner M, Veeravalli K, Gon S, Georgiou G, Beckwith J. Functional plasticity of a peroxidase allows evolution of diverse disulfide-reducing pathways. Proc Natl Acad Sci U S A. 2008;105:6735-40 pubmed publisher
    ..Double mutants lacking thioredoxin reductase (trxB) and glutathione reductase (gor) or glutathione biosynthesis (gshA) cannot grow...
  29. Katoh M, Hiratake J, Oda J. ATP-dependent inactivation of Escherichia coli gamma-glutamylcysteine synthetase by L-glutamic acid gamma-monohydroxamate. Biosci Biotechnol Biochem. 1998;62:1455-7 pubmed
  30. Oktiabr skiĭ O, Ushakov V, Muzyka N, Smirnova G. [The role of thiol redox systems in response of Escherichia coli to distant UV irradiation]. Mikrobiologiia. 2009;78:328-33 pubmed
  31. Apontoweil P, Berends W. Glutathione biosynthesis in Escherichia coli K 12. Properties of the enzymes and regulation. Biochim Biophys Acta. 1975;399:1-9 pubmed
    ..The pathway is controlled by feedback inhibition and not by repression. ..
  32. Watanabe K, Yamano Y, Murata K, Kimura A. The nucleotide sequence of the gene for gamma-glutamylcysteine synthetase of Escherichia coli. Nucleic Acids Res. 1986;14:4393-400 pubmed
    ..For transcription, two sets of consensus promoter signals(-10 and -35 regions) overlapping each other were identified. The terminator signal shows the favored stem-loop structure with an adequate free energy delta G = -22.80 kcal/mol. ..
  33. Haller B, Fuchs J. Mapping of trxB, a mutation responsible for reduced thioredoxin reductase activity. J Bacteriol. 1984;159:1060-2 pubmed
    ..The gene order in this region of the E. coli chromosome was found to probably be serC-aroA-trxB. The location of gshA, the structural gene for gamma-glutamylcystein synthetase, relative to srl and recA also was determined.
  34. Chesney J, Eaton J, Mahoney J. Bacterial glutathione: a sacrificial defense against chlorine compounds. J Bacteriol. 1996;178:2131-5 pubmed
    ..5 to 4.0 mol of hypochlorous acid. We conclude that glutathione defends E. coli cells against attack by chlorine compounds and hydrogen peroxide but, in the case of the halogen compounds, does so nonenzymatically and sacrificially. ..
  35. Moore W, Meister A. Enzymatic synthesis of novel glutathione analogs. Anal Biochem. 1987;161:487-93 pubmed
    ..Fluorine-containing analogs may be used for NMR studies. The enzymatically prepared compounds may also be used as intermediates in the chemical synthesis of other analogs of glutathione and glutathione disulfide. ..
  36. McLaggan D, Logan T, Lynn D, Epstein W. Involvement of gamma-glutamyl peptides in osmoadaptation of Escherichia coli. J Bacteriol. 1990;172:3631-6 pubmed
    ..A mutant deficient in glutathione because of an insertion in the gshA gene was unable to grow above 1...
  37. Murata K, Tani K, Kato J, Chibata I. Excretion of glutathione by methylglyoxal-resistant Escherichia coli. J Gen Microbiol. 1980;120:545-7 pubmed
    ..Methylglyoxal resistance appeared to be due to the simultaneous increase in the activities of these two enzyme systems. ..
  38. Latinwo L, Donald C, Ikediobi C, Silver S. Effects of intracellular glutathione on sensitivity of Escherichia coli to mercury and arsenite. Biochem Biophys Res Commun. 1998;242:67-70 pubmed
    ..sensitivity to mercuric cations and arsenite anions was studied in Escherichia coli mutants that lack glutathione (gshA) with or without an additional mutation affecting the osmotregulant trehalose...
  39. Bouter S, Kerklaan P, Zoetemelk C, Mohn G. Biochemical characterization of glutathione-deficient mutants of Escherichia coli K12 and Salmonella strains TA1535 and TA100. Biochem Pharmacol. 1988;37:577-81 pubmed
    ..It is concluded that the present GSH-deficient strains of E. coli and Salmonella are gshA mutants, analogous to those previously described in E. coli...
  40. Gushima H, Miya T, Murata K, Kimura A. Construction of glutathione-producing strains of Escherichia coli B by recombinant DNA techniques. J Appl Biochem. 1983;5:43-52 pubmed
    ..About 5 mg/ml of glutathione was produced by E. coli cells with pGS500 from 80 mM L-glutamate, 20 mM L-cysteine, and 20 mM glycine within 3 h at 37 degrees C. ..