Measurement of strain dependent toxicity in the indene bioconersion using multi-parameter flow cytometry

A Amanullah, Christopher Hewitt, Alvin Nienow, C Lee, M Chartrain, B Buckland, S Drew, J Woodley

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The bioconversion of indene to cis-(1S,2R) indandiol, a potential key intermediate in the synthesis of Merck's HIV protease inhibitor, CRIXIVAN(TM), can be achieved using a Rhodococcus strain. This study using Rhodococcus 124 reports on the application of multiparameter flow cytometry for the measurement of cell physiological properties based on cytoplasmic membrane (CM) integrity and membrane depolarization as indicators of toxic effects of the substrate, indene. Quantification of intact polarized CM, intact depolarized CM and permeabilized CM of a large population of bacterial cells has been conducted using specific intracellular and membrane-binding fluorescent stains. Measurements of oxygen uptake rate (OUR) and optical density (OD) as indicators of metabolic activity and biomass growth, respectively, were also made. Indene concentrations of up to 0.25 g/L (0.037 g indene/g dry cell weight) did not significantly (
Original languageEnglish
Pages (from-to)239-249
Number of pages11
JournalBiotechnology and Bioengineering
Volume80
Issue number3
DOIs
Publication statusPublished - 1 Jan 2002

Keywords

  • HIV protease inhibitor
  • indene bioconversion
  • substrate, product, and by-product toxicity
  • flow cytometry
  • strain selection

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