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Applied and Environmental Microbiology, September 2004, p. 5511-5521, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5511-5521.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Experimental and Theoretical Bases of Specific Affinity, a Cytoarchitecture-Based Formulation of Nutrient Collection Proposed To Supercede the Michaelis-Menten Paradigm of Microbial Kinetics

D. K. Button,^* Betsy Robertson, Elizabeth Gustafson, and Xiaoming Zhao

Institute of Marine Science and Department of Chemistry and Biochemistry, University of Alaska Fairbanks, Fairbanks, Alaska

Received 23 October 2003/ Accepted 10 May 2004

A theory for solute uptake by whole cells was derived with a focus on the ability of oligobacteria to sequester nutrients. It provided a general relationship that was used to obtain the kinetic constants for in situ marine populations in the presence of naturally occurring substrates. In situ affinities of 0.9 to 400 liters g of cells^–1 h^–1 found were up to 10³ times smaller than those from a "Marinobacter arcticus " isolate, but springtime values were greatly increased by warming. Affinities of the isolate for usual polar substrates but not for hydrocarbons were diminished by ionophores. A kinetic curve or Monod plot was constructed from the best available data for cytoarchitectural components of the isolate by using the theory together with concepts and calculations from first principles. The order of effect of these components on specific affinity was membrane potential > cytoplasmic enzyme concentration > cytoplasmic enzyme affinity > permease concentration > area of the permease site > translation coefficient > porin concentration. Component balance was influential as well; a small increase in cytoplasmic enzyme concentration gave a large increase in the effect of permease concentration. The effect of permease concentration on specific affinity was large, while the effect on K_m was small. These results are in contrast to the Michaelis-Menten theory as applied by Monod that has uptake kinetics dependent on the quality of the permease molecules, with K_m as an independent measure of affinity. Calculations demonstrated that most oligobacteria in the environment must use multiple substrates simultaneously to attain sufficient energy and material for growth, a requirement consistent with communities largely comprising few species.

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* Corresponding author. Mailing address: Institute of Marine Science and Department of Chemistry and Biochemistry, University of Alaska Fairbanks, Fairbanks, AK 99775. Phone: (907) 474-7708. Fax: (907) 474-7204. E-mail: dkbutton{at}ims.uaf.edu

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Applied and Environmental Microbiology, September 2004, p. 5511-5521, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5511-5521.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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