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Applied and Environmental Microbiology, November 1998, p. 4467-4476, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
A Small, Dilute-Cytoplasm, High-Affinity, Novel Bacterium
Isolated by Extinction Culture and Having Kinetic Constants
Compatible with Growth at Ambient Concentrations of Dissolved
Nutrients in Seawater
D. K.
Button,1,2,*
Betsy R.
Robertson,1
Paul W.
Lepp,3 and
Thomas M.
Schmidt3
Institute of Marine
Science,1 and
Department of Chemistry
and Biochemistry,2 University of Alaska
Fairbanks, Fairbanks, Alaska 99775, and
Department of
Microbiology, Michigan State University, East Lansing, Michigan
488243
Received 29 June 1998/Accepted 2 September 1998
Dilutions of raw seawater produced a bacterial isolate capable of
extended growth in unamended seawater. Its 2.9-Mb genome size and 40-fg
dry mass were similar to values for many naturally occurring
aquatic organotrophs, but water and DNA comprised a large
portion of this small chemoheterotroph, as compared to
Escherichia coli. The isolate used only a few aromatic
hydrocarbons and acetate, and glucose and amino acid incorporation were
entirely absent, although many membrane and cytoplasmic proteins were
inducible; it was named Cycloclasticus oligotrophus. A
general rate equation that incorporates saturation phenomena into
specific affinity theory is derived. It is used to relate the kinetic
constants for substrate uptake by the isolate to its cellular proteins. The affinity constant KA for toluene was low at
1.3 µg/liter under optimal conditions, similar to those measured in
seawater, and the low value was ascribed to an unknown slow step such
as limitation by a cytoplasmic enzyme; KA
increased with increasing specific affinities. Specific affinities,
a°s, were protocol sensitive, but under
optimal conditions were 47.4 liters/mg of cells/h, the highest reported
in the literature and a value sufficient for growth in seawater at
concentrations sometimes found. Few rRNA operons, few cytoplasmic
proteins, a small genome size, and a small cell size, coupled with a
high a°s and a low solids content and the
ability to grow without intentionally added substrate, are consistent
with the isolation of a marine bacterium with properties typical
of the bulk of those present.
*
Corresponding author. Mailing address: Institute of
Marine Science, University of Alaska, Fairbanks, AK 99775. Phone: (907) 474-7708. Fax: (907) 474-7204. E-mail:
dkbutton{at}ims.alaska.edu.
Applied and Environmental Microbiology, November 1998, p. 4467-4476, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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