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Chromium reduction in Pseudomonas putida.

Y Ishibashi¹, C Cervantes, S Silver

¹Department of Microbiology and Immunology, University of Illinois, College of Medicine, Chicago 60680.

Applied and Environmental Microbiology

|July 1, 1990

Summary

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Pseudomonas putida PRS2000 reduces toxic hexavalent chromium (chromate) to trivalent chromium using soluble enzymes. This chromate reductase activity is heat sensitive and unaffected by sulfate or nitrate.

Area of Science:

Microbiology
Environmental Science
Biochemistry

Background:

Hexavalent chromium (Cr(VI)) is a toxic environmental pollutant.
Bioremediation using microorganisms offers a sustainable solution for chromium contamination.
Pseudomonas putida is known for its metabolic versatility.

Purpose of the Study:

To investigate the reduction of hexavalent chromium (chromate) by Pseudomonas putida PRS2000.
To characterize the enzyme responsible for chromate reduction.
To determine factors affecting the bioremediation efficiency.

Main Methods:

Utilized cell suspensions and cell-free supernatant fluids of Pseudomonas putida PRS2000.
Assayed for chromate reductase activity in different cellular fractions.

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Determined kinetic parameters (Km) and stability of the enzyme.

Tested the effect of sulfate and nitrate on chromate reduction.

Main Results:

Chromate reductase activity was localized in the soluble protein fraction, not the membrane.
The crude enzyme exhibited heat lability.
The Michaelis constant (Km) for chromate reduction was determined to be 40 microM.
Sulfate and nitrate did not inhibit chromate reduction in vitro or with intact cells.

Conclusions:

Pseudomonas putida PRS2000 possesses soluble chromate reductase activity crucial for bioremediation.
The enzyme's heat sensitivity requires consideration for process optimization.
The lack of inhibition by sulfate and nitrate suggests robustness in mixed-waste environments.