We have developed a novel, ultra-high-throughput screening assay for the detection of cellulase activity based on fluorescence-activated cell sorting (FACS) and double emulsion technology. Cellulase activity is detected using a series of coupled enzymes, including hexose oxidase (HOx), which generates hydrogen peroxide from the reducing sugars released by cellulases in the presence of any natural or artificial substrate. The assay can be adapted to suit a microtiter plate format, but the highest throughput is achieved by using FACS to screen high-complexity cellulase clone libraries. Using this approach, we achieved a 12-fold enrichment of positive (cellulase-expressing) cells in cellulase reference libraries after just one sorting round.
TY - JOUR
AU - Ostafe, Raluca
AU - Prodanović, Radivoje
AU - Commandeur, Ulrich
AU - Fischer, Rainer
PY - 2013
UR - https://cherry.chem.bg.ac.rs/handle/123456789/1606
AB - We have developed a novel, ultra-high-throughput screening assay for the detection of cellulase activity based on fluorescence-activated cell sorting (FACS) and double emulsion technology. Cellulase activity is detected using a series of coupled enzymes, including hexose oxidase (HOx), which generates hydrogen peroxide from the reducing sugars released by cellulases in the presence of any natural or artificial substrate. The assay can be adapted to suit a microtiter plate format, but the highest throughput is achieved by using FACS to screen high-complexity cellulase clone libraries. Using this approach, we achieved a 12-fold enrichment of positive (cellulase-expressing) cells in cellulase reference libraries after just one sorting round.
PB - Academic Press Inc Elsevier Science, San Diego
T2 - Analytical Biochemistry
T1 - Flow cytometry-based ultra-high-throughput screening assay for cellulase activity
VL - 435
IS - 1
SP - 93
EP - 98
DO - 10.1016/j.ab.2012.10.043
UR - Kon_2437
ER -
@article{
author = "Ostafe, Raluca and Prodanović, Radivoje and Commandeur, Ulrich and Fischer, Rainer",
year = "2013",
abstract = "We have developed a novel, ultra-high-throughput screening assay for the detection of cellulase activity based on fluorescence-activated cell sorting (FACS) and double emulsion technology. Cellulase activity is detected using a series of coupled enzymes, including hexose oxidase (HOx), which generates hydrogen peroxide from the reducing sugars released by cellulases in the presence of any natural or artificial substrate. The assay can be adapted to suit a microtiter plate format, but the highest throughput is achieved by using FACS to screen high-complexity cellulase clone libraries. Using this approach, we achieved a 12-fold enrichment of positive (cellulase-expressing) cells in cellulase reference libraries after just one sorting round.",
publisher = "Academic Press Inc Elsevier Science, San Diego",
journal = "Analytical Biochemistry",
title = "Flow cytometry-based ultra-high-throughput screening assay for cellulase activity",
volume = "435",
number = "1",
pages = "93-98",
doi = "10.1016/j.ab.2012.10.043",
url = "Kon_2437"
}
Ostafe, R., Prodanović, R., Commandeur, U.,& Fischer, R.. (2013). Flow cytometry-based ultra-high-throughput screening assay for cellulase activity. in Analytical Biochemistry
Academic Press Inc Elsevier Science, San Diego., 435(1), 93-98.
https://doi.org/10.1016/j.ab.2012.10.043
Kon_2437
Ostafe R, Prodanović R, Commandeur U, Fischer R. Flow cytometry-based ultra-high-throughput screening assay for cellulase activity. in Analytical Biochemistry. 2013;435(1):93-98.
doi:10.1016/j.ab.2012.10.043
Kon_2437 .
