Purification and characterization of rhodobactin: A mixed ligand siderophore from Rhodococcus rhodochrous strain OFS

Suraj Dhungana; Ryszard Michalczyk; Hakim Boukhalfa; Joseph G. Lack; Andrew T. Koppisch; Jason M. Fairlee; Mitchell T. Johnson; Christy E. Ruggiero; Seth G. John; Matthew M. Cox; Cindy C. Browder; Jennifer H. Forsythe; Laura A. Vanderberg; Mary P. Neu; Larry E. Hersman

doi:10.1007/s10534-006-9079-y

Purification and characterization of rhodobactin: A mixed ligand siderophore from Rhodococcus rhodochrous strain OFS

Suraj Dhungana, Ryszard Michalczyk, Hakim Boukhalfa, Joseph G. Lack, Andrew T. Koppisch, Jason M. Fairlee, Mitchell T. Johnson, Christy E. Ruggiero, Seth G. John, Matthew M. Cox, Cindy C. Browder, Jennifer H. Forsythe, Laura A. Vanderberg, Mary P. Neu, Larry E. Hersman

Chemistry and Biochemistry

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

26 Scopus citations

Abstract

The siderophore produced by Rhodococcus rhodochrous strain OFS, rhodobactin, was isolated from iron-deficient cultures and purified by a combination of XAD-7 absorptive/partition resin column and semi-preparative HPLC. The siderophore structure was characterized using 1D and 2D ¹H, ¹³C and ¹⁵N NMR techniques (DQFCOSY, TOCSY, NOESY, HSQC and LR-HSQC) and was confirmed using ESI-MS and MS/MS experiments. The structural characterization revealed that the siderophore, rhodobactin, is a mixed ligand hexadentate siderophore with two catecholate and one hydroxamate moieties for iron chelation. We further investigated the effects of Fe concentrations on siderophore production and found that Fe limiting conditions (Fe concentrations from 0.1 μM to 2.0 μM) facilitated siderophore excretion. Our interests lie in the role that siderophores may have in binding metals at mixed contamination sites (containing metals/radionuclides and organics). Given the broad metabolic capacity of this microbe and its Fe scavenging ability, R. rhodochrous OFS may have a competitive advantage over other organisms employed in bioremediation.

Original language	English (US)
Pages (from-to)	853-867
Number of pages	15
Journal	BioMetals
Volume	20
Issue number	6
DOIs	https://doi.org/10.1007/s10534-006-9079-y
State	Published - Dec 2007

Keywords

Catecholate
Hydroxamate
Rhodobactin
Rhodococcus
Siderophore

ASJC Scopus subject areas

Biomaterials
General Biochemistry, Genetics and Molecular Biology
General Agricultural and Biological Sciences
Metals and Alloys

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Dhungana, S., Michalczyk, R., Boukhalfa, H., Lack, J. G., Koppisch, A. T., Fairlee, J. M., Johnson, M. T., Ruggiero, C. E., John, S. G., Cox, M. M., Browder, C. C., Forsythe, J. H., Vanderberg, L. A., Neu, M. P., & Hersman, L. E. (2007). Purification and characterization of rhodobactin: A mixed ligand siderophore from Rhodococcus rhodochrous strain OFS. BioMetals, 20(6), 853-867. https://doi.org/10.1007/s10534-006-9079-y

Dhungana, S, Michalczyk, R, Boukhalfa, H, Lack, JG, Koppisch, AT, Fairlee, JM, Johnson, MT, Ruggiero, CE, John, SG, Cox, MM, Browder, CC, Forsythe, JH, Vanderberg, LA, Neu, MP & Hersman, LE 2007, 'Purification and characterization of rhodobactin: A mixed ligand siderophore from Rhodococcus rhodochrous strain OFS', BioMetals, vol. 20, no. 6, pp. 853-867. https://doi.org/10.1007/s10534-006-9079-y

@article{9b207a02697348e2abd2801251c08141,

title = "Purification and characterization of rhodobactin: A mixed ligand siderophore from Rhodococcus rhodochrous strain OFS",

abstract = "The siderophore produced by Rhodococcus rhodochrous strain OFS, rhodobactin, was isolated from iron-deficient cultures and purified by a combination of XAD-7 absorptive/partition resin column and semi-preparative HPLC. The siderophore structure was characterized using 1D and 2D 1H, 13C and 15N NMR techniques (DQFCOSY, TOCSY, NOESY, HSQC and LR-HSQC) and was confirmed using ESI-MS and MS/MS experiments. The structural characterization revealed that the siderophore, rhodobactin, is a mixed ligand hexadentate siderophore with two catecholate and one hydroxamate moieties for iron chelation. We further investigated the effects of Fe concentrations on siderophore production and found that Fe limiting conditions (Fe concentrations from 0.1 μM to 2.0 μM) facilitated siderophore excretion. Our interests lie in the role that siderophores may have in binding metals at mixed contamination sites (containing metals/radionuclides and organics). Given the broad metabolic capacity of this microbe and its Fe scavenging ability, R. rhodochrous OFS may have a competitive advantage over other organisms employed in bioremediation.",

keywords = "Catecholate, Hydroxamate, Rhodobactin, Rhodococcus, Siderophore",

author = "Suraj Dhungana and Ryszard Michalczyk and Hakim Boukhalfa and Lack, {Joseph G.} and Koppisch, {Andrew T.} and Fairlee, {Jason M.} and Johnson, {Mitchell T.} and Ruggiero, {Christy E.} and John, {Seth G.} and Cox, {Matthew M.} and Browder, {Cindy C.} and Forsythe, {Jennifer H.} and Vanderberg, {Laura A.} and Neu, {Mary P.} and Hersman, {Larry E.}",

note = "Funding Information: Acknowledgements This work was funded through a grant from the US Department of Energy, Office of Science, Office of Biological and Environmental Research, Natural and Accelerated Bioremediation Science Program (NABIR).",

year = "2007",

month = dec,

doi = "10.1007/s10534-006-9079-y",

language = "English (US)",

volume = "20",

pages = "853--867",

journal = "BioMetals",

issn = "0966-0844",

publisher = "Springer Netherlands",

number = "6",

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TY - JOUR

T1 - Purification and characterization of rhodobactin

T2 - A mixed ligand siderophore from Rhodococcus rhodochrous strain OFS

AU - Dhungana, Suraj

AU - Michalczyk, Ryszard

AU - Boukhalfa, Hakim

AU - Lack, Joseph G.

AU - Koppisch, Andrew T.

AU - Fairlee, Jason M.

AU - Johnson, Mitchell T.

AU - Ruggiero, Christy E.

AU - John, Seth G.

AU - Cox, Matthew M.

AU - Browder, Cindy C.

AU - Forsythe, Jennifer H.

AU - Vanderberg, Laura A.

AU - Neu, Mary P.

AU - Hersman, Larry E.

N1 - Funding Information: Acknowledgements This work was funded through a grant from the US Department of Energy, Office of Science, Office of Biological and Environmental Research, Natural and Accelerated Bioremediation Science Program (NABIR).

PY - 2007/12

Y1 - 2007/12

N2 - The siderophore produced by Rhodococcus rhodochrous strain OFS, rhodobactin, was isolated from iron-deficient cultures and purified by a combination of XAD-7 absorptive/partition resin column and semi-preparative HPLC. The siderophore structure was characterized using 1D and 2D 1H, 13C and 15N NMR techniques (DQFCOSY, TOCSY, NOESY, HSQC and LR-HSQC) and was confirmed using ESI-MS and MS/MS experiments. The structural characterization revealed that the siderophore, rhodobactin, is a mixed ligand hexadentate siderophore with two catecholate and one hydroxamate moieties for iron chelation. We further investigated the effects of Fe concentrations on siderophore production and found that Fe limiting conditions (Fe concentrations from 0.1 μM to 2.0 μM) facilitated siderophore excretion. Our interests lie in the role that siderophores may have in binding metals at mixed contamination sites (containing metals/radionuclides and organics). Given the broad metabolic capacity of this microbe and its Fe scavenging ability, R. rhodochrous OFS may have a competitive advantage over other organisms employed in bioremediation.

AB - The siderophore produced by Rhodococcus rhodochrous strain OFS, rhodobactin, was isolated from iron-deficient cultures and purified by a combination of XAD-7 absorptive/partition resin column and semi-preparative HPLC. The siderophore structure was characterized using 1D and 2D 1H, 13C and 15N NMR techniques (DQFCOSY, TOCSY, NOESY, HSQC and LR-HSQC) and was confirmed using ESI-MS and MS/MS experiments. The structural characterization revealed that the siderophore, rhodobactin, is a mixed ligand hexadentate siderophore with two catecholate and one hydroxamate moieties for iron chelation. We further investigated the effects of Fe concentrations on siderophore production and found that Fe limiting conditions (Fe concentrations from 0.1 μM to 2.0 μM) facilitated siderophore excretion. Our interests lie in the role that siderophores may have in binding metals at mixed contamination sites (containing metals/radionuclides and organics). Given the broad metabolic capacity of this microbe and its Fe scavenging ability, R. rhodochrous OFS may have a competitive advantage over other organisms employed in bioremediation.

KW - Catecholate

KW - Hydroxamate

KW - Rhodobactin

KW - Rhodococcus

KW - Siderophore

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SN - 0966-0844

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SP - 853

EP - 867

JO - BioMetals

JF - BioMetals

IS - 6

ER -

Purification and characterization of rhodobactin: A mixed ligand siderophore from Rhodococcus rhodochrous strain OFS

Abstract

Keywords

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