000842291 001__ 842291 000842291 005__ 20210831151938.0 000842291 020__ $$a978-1-4939-7364-4 (print) 000842291 020__ $$a978-1-4939-7366-8 (electronic) 000842291 0247_ $$2doi$$a10.1007/978-1-4939-7366-8_12 000842291 0247_ $$2ISSN$$a1064-3745 000842291 0247_ $$2ISSN$$a1940-6029 000842291 0247_ $$2pmid$$apmid:29086311 000842291 0247_ $$2WOS$$aWOS:000684272200013 000842291 037__ $$aFZJ-2018-00538 000842291 082__ $$a570 000842291 1001_ $$0P:(DE-HGF)0$$aBornscheuer, Uwe T.$$b0$$eEditor 000842291 245__ $$aHigh-Throughput Screening Assays for Lipolytic Enzymes 000842291 260__ $$aNew York, NY$$bSpringer New York$$c2018 000842291 29510 $$aProtein Engineering / Bornscheuer, Uwe T. (Editor) ; New York, NY : Springer New York, 2018, Chapter 12 ; ISSN: 1064-3745=1940-6029 ; ISBN: 978-1-4939-7364-4=978-1-4939-7366-8 ; doi:10.1007/978-1-4939-7366-8 000842291 300__ $$a209 - 231 000842291 3367_ $$2ORCID$$aBOOK_CHAPTER 000842291 3367_ $$07$$2EndNote$$aBook Section 000842291 3367_ $$2DRIVER$$abookPart 000842291 3367_ $$2BibTeX$$aINBOOK 000842291 3367_ $$2DataCite$$aOutput Types/Book chapter 000842291 3367_ $$0PUB:(DE-HGF)7$$2PUB:(DE-HGF)$$aContribution to a book$$bcontb$$mcontb$$s1548141610_14471 000842291 4900_ $$aMethods in Molecular Biology$$v1685 000842291 520__ $$aScreening 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. 000842291 536__ $$0G:(DE-HGF)POF3-581$$a581 - Biotechnology (POF3-581)$$cPOF3-581$$fPOF III$$x0 000842291 588__ $$aDataset connected to CrossRef Book Series 000842291 7001_ $$0P:(DE-HGF)0$$aHöhne, Matthias$$b1$$eEditor 000842291 7001_ $$0P:(DE-Juel1)143642$$aFulton, Alexander$$b2 000842291 7001_ $$0P:(DE-HGF)0$$aHayes, Marc R.$$b3 000842291 7001_ $$0P:(DE-HGF)0$$aSchwaneberg, Ulrich$$b4 000842291 7001_ $$0P:(DE-Juel1)128906$$aPietruszka, Jörg$$b5$$ufzj 000842291 7001_ $$0P:(DE-Juel1)131457$$aJaeger, Karl-Erich$$b6$$ufzj 000842291 773__ $$a10.1007/978-1-4939-7366-8_12 000842291 909CO $$ooai:juser.fz-juelich.de:842291$$pVDB 000842291 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)128906$$aForschungszentrum Jülich$$b5$$kFZJ 000842291 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)131457$$aForschungszentrum Jülich$$b6$$kFZJ 000842291 9131_ $$0G:(DE-HGF)POF3-581$$1G:(DE-HGF)POF3-580$$2G:(DE-HGF)POF3-500$$3G:(DE-HGF)POF3$$4G:(DE-HGF)POF$$aDE-HGF$$bKey Technologies$$lKey Technologies for the Bioeconomy$$vBiotechnology$$x0 000842291 9141_ $$y2018 000842291 915__ $$0StatID:(DE-HGF)0300$$2StatID$$aDBCoverage$$bMedline 000842291 915__ $$0StatID:(DE-HGF)0310$$2StatID$$aDBCoverage$$bNCBI Molecular Biology Database 000842291 915__ $$0StatID:(DE-HGF)0200$$2StatID$$aDBCoverage$$bSCOPUS 000842291 9201_ $$0I:(DE-Juel1)IMET-20090612$$kIMET$$lInstitut für Molekulare Enzymtechnologie (HHUD)$$x0 000842291 9201_ $$0I:(DE-Juel1)IBOC-20090406$$kIBOC$$lInstitut für Bioorganische Chemie (HHUD)$$x1 000842291 9201_ $$0I:(DE-Juel1)IBG-1-20101118$$kIBG-1$$lBiotechnologie$$x2 000842291 980__ $$acontb 000842291 980__ $$aVDB 000842291 980__ $$aI:(DE-Juel1)IMET-20090612 000842291 980__ $$aI:(DE-Juel1)IBOC-20090406 000842291 980__ $$aI:(DE-Juel1)IBG-1-20101118 000842291 980__ $$aUNRESTRICTED