Flow cytometry-based ultra-high-throughput screening assay for cellulase activity
Само за регистроване кориснике
2013
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
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.
Кључне речи:
Cellulase / Fluorescence assay / Hexose oxidase / Vanadium bromoperoxidase / ViPer / FACS / Double emulsionsИзвор:
Analytical Biochemistry, 2013, 435, 1, 93-98Издавач:
- Academic Press Inc Elsevier Science, San Diego
Финансирање / пројекти:
- Excellence Initiative by the German federal government
Напомена:
- Supplementary material: http://cherry.chem.bg.ac.rs/handle/123456789/3494
DOI: 10.1016/j.ab.2012.10.043
ISSN: 0003-2697
PubMed: 23146590
WoS: 000315477600014
Scopus: 2-s2.0-84873985413
Колекције
Институција/група
Hemijski fakultet / Faculty of ChemistryTY - 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 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" }
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
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 .
Ostafe, Raluca, Prodanović, Radivoje, Commandeur, Ulrich, Fischer, Rainer, "Flow cytometry-based ultra-high-throughput screening assay for cellulase activity" in Analytical Biochemistry, 435, no. 1 (2013):93-98, https://doi.org/10.1016/j.ab.2012.10.043 . .