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Alcaligenes eutrophus as a bacterial chromate sensor

N Peitzsch¹, G Eberz, D H Nies

¹Institut für Mikrobiologie, Halle, Germany.

Applied and Environmental Microbiology

|February 17, 1998

Summary

Researchers developed metal-sensing bacterial strains using plasmid pMOL28. These strains can detect chromate and other heavy metals, paving the way for environmental monitoring applications.

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Area of Science:

Environmental microbiology
Molecular biology
Biotechnology

Background:

The plasmid pMOL28 in Alcaligenes eutrophus CH34 contains adjacent determinants for inducible resistance to chromate (chr) and cobalt/nickel (cnr).
Developing bacterial biosensors requires understanding and manipulating these metal resistance determinants.

Purpose of the Study:

To engineer bacterial strains capable of sensing specific heavy metals.
To investigate the induction patterns of chromate (chr) and cobalt/nickel (cnr) resistance determinants.
To create a functional chromate biosensor using a lux reporter system.

Main Methods:

Mutagenesis of a cloned pMOL28 fragment containing chr and cnr determinants with Tn5-lacZ.
Construction of a broad-host-range IncP1 plasmid (pEBZ141) with a chr::lux transcriptional fusion.
Cultivation of the engineered strain (A. eutrophus AE104(pEBZ141)) in glycerol and testing induction with various metal ions.

Main Results:

The chr::lacZ fusions were specifically induced by chromium.
The cnr determinant showed broad induction by Ni2+, Co2+, Mn2+, chromate, Cu2+, Cd2+, and Zn2+.
The chr::lux fusion in A. eutrophus AE104(pEBZ141) functioned as a chromate biosensor, with optimal induction by chromate and bichromate.
Interactions between chr induction, chromate reduction/accumulation, and sulfate concentration were observed.

Conclusions:

Engineered bacterial strains can serve as effective biosensors for specific heavy metals like chromate.
The chr and cnr determinants exhibit distinct and overlapping induction profiles.
Environmental factors, such as sulfate concentration, can influence metal resistance and biosensor performance.

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