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2005 | 52 | 1 |

Tytuł artykułu

Overexpression of genes involved in phytochelatin biosynthesis in Escherichia coli: effects on growth, cadmium accumulation and thiol level

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
In Escherichia coli, heterologous production of Schizosaccharomyces pombe phytochelatin synthase (PCS) along with overproduction of E. coli serine acetyltransferase (SAT) and gamma-glutamylcysteine synthase (gammaECS) was achieved and resulted in the accumulation of phytochelatins in bacterial cells. Overproduction of either gammaECS alone or simultaneous production of all three proteins in bacterial cells were accompanied by reduced growth rate in liquid cultures. Interestingly, bacteria overproducing either gammaECS or both SAT and gammaECS (with elevated level of gamma-glutamylcysteine but not of phytochelatins) were able to accumulate more cadmium per dry weight than the control. However, the most efficient cadmium accumulation was observed in bacteria with elevated levels of all three proteins: SAT, gammaECS and PCS. Therefore, “pushing” the entire pathway might be the most promising approach in modification of bacteria for potential bioremediation purposes because the level of intermediates, cysteine and glutathione, can limit the rate of production of phytochelatins. However, in such bacteria other metabolic process might become limiting for efficient growth.

Wydawca

-

Rocznik

Tom

52

Numer

1

Opis fizyczny

p.109-116,fig.,ref.

Twórcy

  • Polish Academy of Sciences, A.Pawinskiego 5A, 02-106 Warsaw, Poland
autor
autor
autor
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autor

Bibliografia

  • Błaszczyk A. (2004) Modification of sulfur metabolism in tobacco and identification of genes regulated by sulfur starvation. PhD Thesis, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw.
  • Chen J, Zhou J, Goldsbrough PB. (1997) Characterization of phytochelatins synthase from tomato. Physiol Plant.; 101: 165–72.
  • Clemens S, Kim EJ, Neumann D, Schroeder JI. (1999) Tolerance to toxic metals by a gene family of phytochelatin synthases from plants and yeast. EMBO J.; 18: 3325–33.
  • Grill E, Loffler S, Winnacker E-L, Zenk M. (1989) Phytochelatins, the heavy metal binding peptides of plants, are synthesized from glutathione by a specific gamma-glutamylcysteine dipeptidyl transpeptidase (phytochelatins synthase). Proc Natl Acad Sci USA.; 86: 6838–42.
  • Heiss S, Schafer HJ, Haag-Kerwer A, Rausch T. (1999) Cloning sulfur assimilation genes of Brassica juncea L.: cadmium differentially affects the expression of a putative low-affinity sulfate transporter and isoforms of ATP sulfurylase and APS reductase. Plant Mol Biol.; 39: 847–57.
  • Klapheck S, Schlunz S, Bergmann L. (1995) Synthesis of phytochelatins and homo-phytochelatins in Pisum sativum L. Plant Physiol.; 107: 515–21.
  • Lee M, Leustek T. (1999) Identification of the gene encoding homoserine kinase from Arabidopsis thaliana and characterization of the recombinant enzyme derived from the gene. Arch Biochem Biophys.; 372: 135–42.
  • Lee S, Moon JS, Ko TS, Petros D, Goldsbrough PB, Korban SS. (2003) Overexpression of Arabidopsis phytochelatin synthase paradoxi- cally leads to hypersensitivity to cadmium stress. Plant Physiol.; 131: 656–63.
  • Li Y, Wei G, Chen J. (2004) Glutathione: a review on biotechnological production. Appl Microbiol Biotechnol.; 66: 233–42.
  • Rauser WE. (1999) Structure and function of metal chelators produced by plants: the case for organic acids, amino acids, phytin, and metallothioneins. Cell Biochem Biophys.; 31: 19–48.
  • Sambrook J, Fritsch EF, Maniatis T. (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, Cold Spring Harbor Laboratory Press.
  • Sanita di Toppi LS, Gabbrielli R. (1999) Response to cadmium in higher plants. Environ Exp Bot.; 41: 105–30.
  • Sauge-Merle S, Cuine S, Carrier P, Lecomte-Pradines C, Luu DT, Peltier G. (2003) Enhanced toxic metal accumulation in engineered bacterial cells expressing Arabidopsis thaliana phytochelatin synthase. Appl Environ Microbiol.; 69: 490–4.
  • Schafer HJ, Haag-Kerwer A, Rausch T. (1998)
  • cDNA cloning and expression analysis of genes encoding GSH synthesis in roots of the heavy-metal accumulator Brassica juncea L.: evidence for Cd-induction of a putative mitochondrial gamma-glutamylcysteine synthetase isoform. Plant Mol Biol.; 37: 87–97.
  • Zenk MH. (1996) Heavy metal detoxification in higher plants — a review. Gene.; 179: 21–30.

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

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