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Activity and stability of a recombinant plasmid‐borne TCE degradative pathway in suspended cultures
Author(s) -
Sharp Robert R.,
Bryers James D.,
Jones Warren G.,
Shields Malcolm S.
Publication year - 1998
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19980205)57:3<287::aid-bit5>3.0.co;2-d
Subject(s) - plasmid , growth rate , microbiology and biotechnology , kinetics , burkholderia , growth inhibition , biology , bacterial growth , biodegradation , recombinant dna , chemistry , bacteria , biochemistry , in vitro , gene , genetics , ecology , physics , geometry , mathematics , quantum mechanics
The retention and expression of the plasmid‐borne, TCE degradative toluene‐ ortho ‐monooxygenase (TOM) pathway in suspended continuous cultures of transconjugant Burkholderia cepacia 17616 (TOM 31c ) were studied. Acetate growth and TCE degradation kinetics for the transconjugant host are described and utilized in a plasmid loss model. Plasmid maintenance did not have a significant effect on the growth rate of the transconjugant. Both plasmid‐bearing and plasmid‐free strains followed Andrews inhibition growth kinetics when grown on acetate and had maximum growth rates of 0.22 h −1 . The transconjugant was capable of degrading TCE at a maximum rate of 9.7 nmol TCE/min · mg protein, which is comparable to the rates found for the original plasmid host, Burkholderia cepacia PR1 31 (TOM 31c ). The specific activity of the TOM pathway was found to be a linear function of growth rate. Plasmid maintenance was studied at three different growth rates: 0.17/h, 0.1/h, and 0.065/h. Plasmid maintenance was found to be a function of growth rate, with the probability of loss ranging from 0.027 at a growth rate of 0.065/h to 0.034 at a growth rate 0.17/h. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 287–296, 1998.