Cloning, purification and characterization of Geobacillus stearothermophilus V uroporphyrinogen-III C-methyltransferase: evaluation of its role in resistance to potassium tellurite in Escherichia coli

Manuel A. Araya; Juan C. Tantaleán; José M. Pérez; Derie E. Fuentes; Iván L. Calderón; Claudia P. Saavedra; Radhika Burra; Thomas G. Chasteen; Claudio C. Vásquez

doi:10.1016/j.resmic.2008.12.004

Cloning, purification and characterization of Geobacillus stearothermophilus V uroporphyrinogen-III C-methyltransferase: evaluation of its role in resistance to potassium tellurite in Escherichia coli

Manuel A. Araya, Juan C. Tantaleán, José M. Pérez, Derie E. Fuentes, Iván L. Calderón, Claudia P. Saavedra, Radhika Burra, Thomas G. Chasteen, Claudio C. Vásquez

CIENCIAS BIOLÓGICAS

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7 Scopus citations

Abstract

The Geobacillus stearothermophilus V cobA gene encoding uroporphyrinogen-III C-methyltransferase (also referred to as SUMT) was cloned into Escherichia coli and the recombinant enzyme was overexpressed and purified to homogeneity. The enzyme binds S-adenosyl-l-methionine and catalyzes the production of III methyl uroporphyrinogen in vitro. E. coli cells expressing the G. stearothermophilus V cobA gene exhibited increased resistance to potassium tellurite and potassium tellurate. Site-directed mutagenesis of cobA abolished tellurite resistance of the mesophilic, heterologous host and SUMT activity in vitro. No methylated, volatile derivatives of tellurium were found in the headspace of tellurite-exposed cobA-expressing E. coli, suggesting that the role of SUMT methyltransferase in tellurite(ate) detoxification is not related to tellurium volatilization.

Original language	English
Pages (from-to)	125-133
Number of pages	9
Journal	Research in Microbiology
Volume	160
Issue number	2
DOIs	https://doi.org/10.1016/j.resmic.2008.12.004
State	Published - Mar 2009

Bibliographical note

Funding Information:
This work received financial support from Fondecyt grant # 1060022 and Dicyt-USACH to C.C.V. and from the Robert A. Welch Foundation (X-011) at Sam Houston State University to T.G.C. and R.B.

Funding Information:
M.A.A. and D.E.F. were supported by doctoral fellowships from Mecesup, Chile. I.L.C. received a doctoral fellowship from Conicyt, Chile and J.M.P. was supported by Mecesup, Conicyt and Dicyt-USACH, Chile.

Keywords

Methyltransferase
S-adenosyl-l-methionine
SUMT
Tellurite
Uroporphyrinogen
cobA

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10.1016/j.resmic.2008.12.004

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Araya, M. A., Tantaleán, J. C., Pérez, J. M., Fuentes, D. E., Calderón, I. L., Saavedra, C. P., Burra, R., Chasteen, T. G., & Vásquez, C. C. (2009). Cloning, purification and characterization of Geobacillus stearothermophilus V uroporphyrinogen-III C-methyltransferase: evaluation of its role in resistance to potassium tellurite in Escherichia coli. Research in Microbiology, 160(2), 125-133. https://doi.org/10.1016/j.resmic.2008.12.004

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title = "Cloning, purification and characterization of Geobacillus stearothermophilus V uroporphyrinogen-III C-methyltransferase: evaluation of its role in resistance to potassium tellurite in Escherichia coli",

abstract = "The Geobacillus stearothermophilus V cobA gene encoding uroporphyrinogen-III C-methyltransferase (also referred to as SUMT) was cloned into Escherichia coli and the recombinant enzyme was overexpressed and purified to homogeneity. The enzyme binds S-adenosyl-l-methionine and catalyzes the production of III methyl uroporphyrinogen in vitro. E. coli cells expressing the G. stearothermophilus V cobA gene exhibited increased resistance to potassium tellurite and potassium tellurate. Site-directed mutagenesis of cobA abolished tellurite resistance of the mesophilic, heterologous host and SUMT activity in vitro. No methylated, volatile derivatives of tellurium were found in the headspace of tellurite-exposed cobA-expressing E. coli, suggesting that the role of SUMT methyltransferase in tellurite(ate) detoxification is not related to tellurium volatilization.",

keywords = "Methyltransferase, S-adenosyl-l-methionine, SUMT, Tellurite, Uroporphyrinogen, cobA",

author = "Araya, {Manuel A.} and Tantale{\'a}n, {Juan C.} and P{\'e}rez, {Jos{\'e} M.} and Fuentes, {Derie E.} and Calder{\'o}n, {Iv{\'a}n L.} and Saavedra, {Claudia P.} and Radhika Burra and Chasteen, {Thomas G.} and V{\'a}squez, {Claudio C.}",

note = "Funding Information: This work received financial support from Fondecyt grant # 1060022 and Dicyt-USACH to C.C.V. and from the Robert A. Welch Foundation (X-011) at Sam Houston State University to T.G.C. and R.B. Funding Information: M.A.A. and D.E.F. were supported by doctoral fellowships from Mecesup, Chile. I.L.C. received a doctoral fellowship from Conicyt, Chile and J.M.P. was supported by Mecesup, Conicyt and Dicyt-USACH, Chile. ",

year = "2009",

month = mar,

doi = "10.1016/j.resmic.2008.12.004",

language = "Ingl{\'e}s",

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Araya, MA, Tantaleán, JC, Pérez, JM, Fuentes, DE, Calderón, IL, Saavedra, CP, Burra, R, Chasteen, TG & Vásquez, CC 2009, 'Cloning, purification and characterization of Geobacillus stearothermophilus V uroporphyrinogen-III C-methyltransferase: evaluation of its role in resistance to potassium tellurite in Escherichia coli', Research in Microbiology, vol. 160, no. 2, pp. 125-133. https://doi.org/10.1016/j.resmic.2008.12.004

Cloning, purification and characterization of Geobacillus stearothermophilus V uroporphyrinogen-III C-methyltransferase: evaluation of its role in resistance to potassium tellurite in Escherichia coli. / Araya, Manuel A.; Tantaleán, Juan C.; Pérez, José M. et al.
In: Research in Microbiology, Vol. 160, No. 2, 03.2009, p. 125-133.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Cloning, purification and characterization of Geobacillus stearothermophilus V uroporphyrinogen-III C-methyltransferase

T2 - evaluation of its role in resistance to potassium tellurite in Escherichia coli

AU - Araya, Manuel A.

AU - Tantaleán, Juan C.

AU - Pérez, José M.

AU - Fuentes, Derie E.

AU - Calderón, Iván L.

AU - Saavedra, Claudia P.

AU - Burra, Radhika

AU - Chasteen, Thomas G.

AU - Vásquez, Claudio C.

N1 - Funding Information: This work received financial support from Fondecyt grant # 1060022 and Dicyt-USACH to C.C.V. and from the Robert A. Welch Foundation (X-011) at Sam Houston State University to T.G.C. and R.B. Funding Information: M.A.A. and D.E.F. were supported by doctoral fellowships from Mecesup, Chile. I.L.C. received a doctoral fellowship from Conicyt, Chile and J.M.P. was supported by Mecesup, Conicyt and Dicyt-USACH, Chile.

PY - 2009/3

Y1 - 2009/3

N2 - The Geobacillus stearothermophilus V cobA gene encoding uroporphyrinogen-III C-methyltransferase (also referred to as SUMT) was cloned into Escherichia coli and the recombinant enzyme was overexpressed and purified to homogeneity. The enzyme binds S-adenosyl-l-methionine and catalyzes the production of III methyl uroporphyrinogen in vitro. E. coli cells expressing the G. stearothermophilus V cobA gene exhibited increased resistance to potassium tellurite and potassium tellurate. Site-directed mutagenesis of cobA abolished tellurite resistance of the mesophilic, heterologous host and SUMT activity in vitro. No methylated, volatile derivatives of tellurium were found in the headspace of tellurite-exposed cobA-expressing E. coli, suggesting that the role of SUMT methyltransferase in tellurite(ate) detoxification is not related to tellurium volatilization.

AB - The Geobacillus stearothermophilus V cobA gene encoding uroporphyrinogen-III C-methyltransferase (also referred to as SUMT) was cloned into Escherichia coli and the recombinant enzyme was overexpressed and purified to homogeneity. The enzyme binds S-adenosyl-l-methionine and catalyzes the production of III methyl uroporphyrinogen in vitro. E. coli cells expressing the G. stearothermophilus V cobA gene exhibited increased resistance to potassium tellurite and potassium tellurate. Site-directed mutagenesis of cobA abolished tellurite resistance of the mesophilic, heterologous host and SUMT activity in vitro. No methylated, volatile derivatives of tellurium were found in the headspace of tellurite-exposed cobA-expressing E. coli, suggesting that the role of SUMT methyltransferase in tellurite(ate) detoxification is not related to tellurium volatilization.

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KW - S-adenosyl-l-methionine

KW - SUMT

KW - Tellurite

KW - Uroporphyrinogen

KW - cobA

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U2 - 10.1016/j.resmic.2008.12.004

DO - 10.1016/j.resmic.2008.12.004

M3 - Artículo

C2 - 19154787

AN - SCOPUS:60949089621

SN - 0923-2508

VL - 160

SP - 125

EP - 133

JO - Research in Microbiology

JF - Research in Microbiology

IS - 2

ER -

Cloning, purification and characterization of Geobacillus stearothermophilus V uroporphyrinogen-III C-methyltransferase: evaluation of its role in resistance to potassium tellurite in Escherichia coli

Abstract

Bibliographical note

Keywords

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