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@INBOOK{Bornscheuer:842291,
author = {Fulton, Alexander and Hayes, Marc R. and Schwaneberg,
Ulrich and Pietruszka, Jörg and Jaeger, Karl-Erich},
editor = {Bornscheuer, Uwe T. and Höhne, Matthias},
title = {{H}igh-{T}hroughput {S}creening {A}ssays for {L}ipolytic
{E}nzymes},
volume = {1685},
address = {New York, NY},
publisher = {Springer New York},
reportid = {FZJ-2018-00538},
isbn = {978-1-4939-7364-4 (print)},
series = {Methods in Molecular Biology},
pages = {209 - 231},
year = {2018},
comment = {Protein Engineering / Bornscheuer, Uwe T. (Editor) ; New
York, NY : Springer New York, 2018, Chapter 12 ; ISSN:
1064-3745=1940-6029 ; ISBN:
978-1-4939-7364-4=978-1-4939-7366-8 ;
doi:10.1007/978-1-4939-7366-8},
booktitle = {Protein Engineering / Bornscheuer, Uwe
T. (Editor) ; New York, NY : Springer
New York, 2018, Chapter 12 ; ISSN:
1064-3745=1940-6029 ; ISBN:
978-1-4939-7364-4=978-1-4939-7366-8 ;
doi:10.1007/978-1-4939-7366-8},
abstract = {Screening is defined as the identification of hits within a
large library of variants of an enzyme or protein with a
predefined property. In theory, each variant present in the
respective library needs to be assayed; however, to save
time and consumables, many screening regimes involve a
primary round to identify clones producing active enzymes.
Such primary or prescreenings for lipolytic enzyme activity
are often carried out on agar plates containing pH
indicators or substrates as triolein or tributyrin.
Subsequently, high-throughput screening assays are usually
performed in microtiter plate (MTP) format using chromogenic
or fluorogenic substrates and, if available, automated
liquid handling robotics. Here, we describe different assay
systems to determine the activity and enantioselectivity of
lipases and esterases as well as the synthesis of several
substrates. We also report on the construction of a complete
site saturation library derived from lipase A of Bacillus
subtilis and its testing for detergent tolerance. This
approach allows for the identification of amino acids
affecting sensitivity or resistance against different
detergents.},
cin = {IMET / IBOC / IBG-1},
ddc = {570},
cid = {I:(DE-Juel1)IMET-20090612 / I:(DE-Juel1)IBOC-20090406 /
I:(DE-Juel1)IBG-1-20101118},
pnm = {581 - Biotechnology (POF3-581)},
pid = {G:(DE-HGF)POF3-581},
typ = {PUB:(DE-HGF)7},
pubmed = {pmid:29086311},
UT = {WOS:000684272200013},
doi = {10.1007/978-1-4939-7366-8_12},
url = {https://juser.fz-juelich.de/record/842291},
}