Improved rapid sampling for in vivo kinetics of intracellular metabolites in Saccharomyces cerevisiae.

Abstract:

:An integrated approach is used to develop a rapid sampling strategy for the quantitative analysis of in vivo kinetic behavior based on measured concentrations of intracellular metabolites in Saccharomyces cerevisiae. Emphasis is laid on small sample sizes during sampling and analysis. Subsecond residence times are accomplished by minimizing the dead volume of the sterile sampling system and by maximizing flow rates through application of vacuum to the sampling tubes in addition to the overpressure in the fermenter. A specially designed sample tube adapter facilitates sampling intervals of 4 to 5 s for various test tube types. Statistical analysis of the results obtained from enzymatic and liquid chromatography mass spectrometry (LC-MSMS) analysis of the metabolite concentrations was used to optimize the sampling protocol. The most notable improvement is reached through the introduction of vacuum drying of the cell extract. The presented system is capable of reliably dealing with fermenter samples as small as 1-g with a variation of less than 3%, and is thus ideally suited for intracellular measurements on small, lab-scale fermenters.

journal_name

Biotechnol Bioeng

authors

Lange HC,Eman M,van Zuijlen G,Visser D,van Dam JC,Frank J,de Mattos MJ,Heijnen JJ

doi

10.1002/bit.10048

subject

Has Abstract

pub_date

2001-11-20 00:00:00

pages

406-15

issue

4

eissn

0006-3592

issn

1097-0290

pii

10.1002/bit.10048

journal_volume

75

pub_type

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