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An alternate method for estimation of cell growth kinetics from batch cultures
Author(s) -
Lee Jeewon,
Chang HsiaoLung,
Parulekar Satish J.,
Hong Juan
Publication year - 1991
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/bit.260370106
Subject(s) - bacillus subtilis , plasmid , exponential growth , recombinant dna , kinetics , strain (injury) , microorganism , reversion , biology , cell growth , bacterial growth , biological system , cell culture , chemistry , microbiology and biotechnology , bacteria , biochemistry , genetics , dna , mathematics , phenotype , gene , mathematical analysis , physics , anatomy , quantum mechanics
An alternative to estimation of cell growth kinetics via continuous culture experiments is proposed in this article. The method employed is based on batch culture experiments with very small inocula (initial cell concentrations being typically less than 5000 cells/mL). Such low initial cell concentrations result in extended exponential cell growth phase during which culture conditions remain unchanged, thereby permitting precise estimation of specific cell growth rates from batch experiments especially for fast‐growing microorganisms such as Bacillus species. The effectiveness and utility of this approach are demonstrated via several experiments conducted with a wild‐type strain ( Bacillus subtilis TN106) and a recombinant strain ( B. subtilis TN106[pAT5]). True establishment of exponential growth phase requires insignificant variance of most of the culture conditions during the initial growth phase. Satisfaction of this requirement is demonstrated for microbial systems investigated here. This approach is especially well suited for recombinant microorganisms containing segregationally unstable plasmids, since estimation of growth kinetics of these from continuous cultures is very difficult and highly unreliable due to continual reversion of recombinant ceils to plasmid‐free host cells unless some selection pressure is applied at levels sufficient to keep the presence of plasmid‐free cells minimal.

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