Abstract:
:Multi-parameter flow cytometric techniques coupled with dual colour fluorescent staining were used to study the physical and metabolic consequences of inclusion body formation in batch cultures of the recombinant Escherichia coli strain MSD3735. This strain contains a plasmid coding for the isopropylthiogalactopyranoside-inducible model eukaryotic protein AP50. It is known that the synthesis of foreign proteins at high concentrations can exert a severe metabolic stress on the host cell and that morphological changes can occur. In this work, using various points of induction, it was shown that inclusion body formation is followed immediately by measurable changes in the characteristic intrinsic light scatter patterns for the individual cell (forward scatter, 90 degrees side scatter) and a concomitant progressive change in the individual cell physiological state with respect to both cytoplasmic membrane polarisation and permeability. This work establishes flow cytometry as a potentially valuable tool for monitoring recombinant fermentation processes, providing important information for scale-up. Further, we discuss the possibility of optimising inclusion body formation by manipulating the fermentation conditions based on these rapid "real-time" measurements.
journal_name
J Ind Microbiol Biotechnoljournal_title
Journal of industrial microbiology & biotechnologyauthors
Lewis G,Taylor IW,Nienow AW,Hewitt CJdoi
10.1007/s10295-004-0151-8subject
Has Abstractpub_date
2004-08-01 00:00:00pages
311-22issue
7eissn
1367-5435issn
1476-5535journal_volume
31pub_type
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journal_title:Journal of industrial microbiology & biotechnology
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journal_title:Journal of industrial microbiology & biotechnology
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journal_title:Journal of industrial microbiology & biotechnology
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journal_title:Journal of industrial microbiology & biotechnology
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journal_title:Journal of industrial microbiology & biotechnology
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更新日期:2011-01-01 00:00:00
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journal_title:Journal of industrial microbiology & biotechnology
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更新日期:2013-09-01 00:00:00
abstract::Sphingomonas xenophaga BN6 was isolated from the river Elbe as a member of a multispecies bacterial culture which mineralized 6-aminonaphthalene-2-sulfonate. Pure cultures of strain BN6 converted a wide range of amino- and hydroxynaphthalene-2-sulfonates via a catabolic pathway similar to that described for the metabo...
journal_title:Journal of industrial microbiology & biotechnology
pub_type: 杂志文章
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更新日期:1999-10-01 00:00:00
abstract::The industrial crops of microalgae use processes calling upon the presence of parts of metal nature such as steel 316L type. The goal of this study is to test the electrochemical behavior of this material in a marine culture of microalgae. Porphyridium purpureum was used under a photoperiod of alternation darkness/lig...
journal_title:Journal of industrial microbiology & biotechnology
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更新日期:2011-12-01 00:00:00
abstract::p-Anisaldehyde (4-methoxybenzaldehyde), an extract from Pimpinella anisum L. seeds, is a potential novel preservative. To reveal the possible action mechanism of p-anisaldehyde against microorganisms, yeast-based commercial oligonucleotide microarrays were used to analyze the genome-wide transcriptional changes in res...
journal_title:Journal of industrial microbiology & biotechnology
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