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Applied and Environmental Microbiology, August 2000, p. 3408-3414, Vol. 66, No. 8
Department of Microbiology, Swiss Federal
Institute for Environmental Science and Technology (EAWAG),
Dübendorf,1 and Institute of
Biotechnology, Swiss Federal Institute of Technology (ETH),
Zürich,2 Switzerland
Received 14 April 2000/Accepted 30 May 2000
Pseudomonas oleovorans ATCC 29347 was grown in
chemostat culture at different dilution rates with mineral media
varying in their ratios of octanoate to ammonia
(C0/N0 ratio). At all dilution rates tested,
three distinct growth regimes were observed: (i) carbon limitation with
NH4+ in excess at low
C0/N0 ratios, (ii) purely nitrogen-limited
growth conditions at high C0/N0 ratios with
residual octanoate in the culture supernatant, and (iii) an
intermediate zone of dual-nutrient-limited growth conditions where both
the concentration of octanoate and that of ammonia were very low. The
dual-nutrient-limited growth zone shifted to higher
C0/N0 ratios with decreasing dilution rates, and the extension of the dual-nutrient-limited growth zone was inversely proportional to the growth rate. The cells accumulated the
storage compound medium-chain-length
poly[(R)-3-hydroxyalkanoate] (mcl-PHA) during dual (C and
N)-nutrient-limited and N-limited growth conditions. Within the
dual-nutrient-limited growth zone, the cellular mcl-PHA contents
increased when the C0/N0 ratio in the feed was
increased, whereas the cellular mcl-PHA level was independent from the
feed C0/N0 ratio during N-limited growth. The
monomeric composition of the accumulated mcl-PHA was independent of
both the dilution rate and the feed C0/N0 ratio
and consisted of 12 mol% 3-hydroxyhexanoic acid and 88 mol%
3-hydroxyoctanoic acid. Accumulation of mcl-PHA led to an increase in
the cellular C/N ratio and to changes in elemental growth yields for
nitrogen and carbon.
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Accumulation of
Poly[(R)-3-Hydroxyalkanoates] in Pseudomonas
oleovorans during Growth with Octanoate in Continuous Culture at
Different Dilution Rates
*
Corresponding author. Mailing address: Department of
Microbiology, EAWAG, Überlandstrasse 133, CH-8600
Dübendorf, Switzerland. Phone: 41 1 823 51 58. Fax: 41 1 823 55 47. E-mail: egli{at}eawag.ch.
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