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000782247 1001_ $$aMinges, Hannah.
000782247 24510 $$aEngineering of Halogenases towards Synthetic Applications :$$bIncreasing the Thermostability and Investigations on a Marine Brominase Bmp5.
000782247 260__ $$aWiesbaden :$$bSpringer Fachmedien Wiesbaden,$$c2017.
000782247 300__ $$a1 online resource (123 pages).
000782247 336__ $$atext$$btxt$$2rdacontent
000782247 337__ $$acomputer$$bc$$2rdamedia
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000782247 4901_ $$aBestMasters
000782247 504__ $$aIncludes bibliographical references.
000782247 5050_ $$6880-01$$aTable of contents; List of figures; List of schemes; List of abbreviations; 1 Introduction; 1.1 Value of biocatalysis in chemical synthesis; 1.2 Natural halogenated compounds; 1.3 Halogenating enzymes; 1.4 The FAD-dependent halogenase family; 1.5 Biocatalytic application of tryptophan halogenases; 1.6 Directed enzyme evolution; 1.7 Halogenated marine metabolites; 1.8 FAD-dependent halogenase Bmp5; 2 Motivation; 3 Material and Methods; 3.1 Analytics; 3.2 Laboratory Equipment; 3.3 Chemicals and solvents; 3.4 Molecular biological methods; 3.5 Biochemical methods; 3.6 Directed evolution.
000782247 5058_ $$a6.1.2 Expression of Bmp5 in E. coli BL21 (DE3) pGro76.1.3 Protein purification via HisTALON affinity chromatography; 6.1.4 Bmp5 expression in E. coli BL21 (DE3); 6.2 Identification of Bmp5; 6.2.1 Detection of His6-tagged Bmp5 by Western blot analysis; 6.2.2 Identification of Bmp5 by peptide mass fingerprint; 6.3 Enzymatic bromination of 4-HBA after purification of His6-Bmp5; 6.4 Generation of GST-tagged Bmp5; 6.4.1 Subcloning of bmp5 into pETM30 vector; 6.4.1.1 Identification of positive pETM30-GST-bmp5 clones by colony-PCR; 6.4.2 Expression and purification of GST-Bmp5.
000782247 5058_ $$a6.4.3 Attempts for enzymatic bromination of 4-HBA after purification of GST-Bmp56.5 Establishment of cofactor regeneration for Bmp5 in E. coli lysate; 6.5.1 Investigation of glucose dehydrogenase; 6.5.2 Optimization of cofactor regeneration in Bmp5-catalyzed bromination employing alcohol dehydrogenase; 6.5.3 Identification of reaction components by GC-MS analysis; 6.5.4 Study of Bmp5's bromination ability in presence of chloride; 6.5.5 Examination of iodination ability; 6.6 Investigation of Bmp5's substrate scope; 6.7 Preparative synthesis of 2,4-dibromophenol using Bmp5-ADHcombiCLEAs.
000782247 5058_ $$a7 Establishment of a marine brominase7.1 Establishment of the brominase Bmp5 for heterologous expression; 7.2 Expression of GST-Bmp5 fusion protein; 7.3 His6-Bmp5 revealed sufficient enzymatic activity after purification; 7.4 Establishment of cofactor regeneration system for Bmp5-catalyzed reactions; 7.5 Evidence of Bmp5 expression by Western blot analysis and peptide mass fingerprint; 7.6 Detected reaction compounds confirmed proposed Bmp5-catalyzed reaction mechanism; 8 Outlook; 9 Summary; Literature.
000782247 506__ $$aAccess limited to authorized users.
000782247 588__ $$aDescription based on print version record.
000782247 650_0 $$aEnzymes$$xBiotechnology.
000782247 77608 $$iPrint version:$$aMinges, Hannah.$$tEngineering of Halogenases towards Synthetic Applications : Increasing the Thermostability and Investigations on a Marine Brominase Bmp5.$$dWiesbaden : Springer Fachmedien Wiesbaden, ©2017$$z9783658184094
000782247 830_0 $$aBestMasters.
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000782247 85640 $$3SpringerLink$$uhttps://univsouthin.idm.oclc.org/login?url=http://link.springer.com/10.1007/978-3-658-18410-0$$zOnline Access$$91397441.1
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