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Applied and Environmental Microbiology, September 2000, p. 4037-4044, Vol. 66, No. 9
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Effect of Oxygen on Formation and Structure of Azotobacter vinelandii Alginate and Its Role in Protecting Nitrogenase

W. Sabra, A.-P. Zeng,* H. Lünsdorf, and W.-D. Deckwer

Biochemical Engineering Division, Gesellschaft für Biotechnologische Forschung mbH, 38124 Braunschweig, Germany

Received 15 February 2000/Accepted 6 June 2000

The activity of nitrogenase in the nitrogen-fixing bacterium Azotobacter vinelandii grown diazotrophically under aerobic conditions is generally considered to be protected against O2 by a high respiration rate. In this work, we have shown that a high rate of respiration is not the prevailing mechanism for nitrogenase protection in A. vinelandii grown in phosphate-limited nitrogen-free chemostat culture. Instead, the formation of alginate appeared to play a decisive role in protecting the nitrogenase that is required for cell growth in this culture. Depending on the O2 tension and cell growth rate, the formation rate and composition of alginate released into the culture broth varied significantly. Furthermore, transmission electron microscopic analysis of cell morphology and the cell surface revealed the existence of an alginate capsule on the surface of A. vinelandii. The composition, thickness, and compactness of this alginate capsule also varied significantly. In general, increasing O2 tension led to the formation of alginate with a higher molecular weight and a greater L-guluronic acid content. The alginate capsule was accordingly thicker and more compact. In addition, the formation of the alginate capsule was found to be strongly affected by the shear rate in a bioreactor. Based on these experimental results, it is suggested that the production of alginate, especially the formation of an alginate capsule on the cell surface, forms an effective barrier for O2 transfer into the cell. It is obviously the quality, not the quantity, of alginate that is decisive for the protection of nitrogenase.

* Corresponding author. Mailing address: GBF---Gesellschaft für Biotechnologische Forschung mbH, Biochemical Engineering Division, Mascheroder Weg 1, D-38124 Braunschweig, Germany. Phone: 49-531-6181-188. Fax: 49-531-6181-751. E-mail: AZE{at}GBF.de.

Applied and Environmental Microbiology, September 2000, p. 4037-4044, Vol. 66, No. 9
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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