TY - JOUR
T1 - Measurement of strain dependent toxicity in the indene bioconersion using multi-parameter flow cytometry
AU - Amanullah, A
AU - Hewitt, Christopher
AU - Nienow, Alvin
AU - Lee, C
AU - Chartrain, M
AU - Buckland, B
AU - Drew, S
AU - Woodley, J
PY - 2002/1/1
Y1 - 2002/1/1
N2 - 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 (
AB - 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 (
KW - HIV protease inhibitor
KW - indene bioconversion
KW - substrate, product, and by-product toxicity
KW - flow cytometry
KW - strain selection
UR - http://www.scopus.com/inward/record.url?scp=0037027383&partnerID=8YFLogxK
U2 - 10.1002/bit.10477
DO - 10.1002/bit.10477
M3 - Article
C2 - 12226855
SN - 1097-0290
VL - 80
SP - 239
EP - 249
JO - Biotechnology and Bioengineering
JF - Biotechnology and Bioengineering
IS - 3
ER -