Screening for Enantioselective Enzymes

Franken, Benjamin; Jaeger, Karl‐Erich; Pietruszka, Jörg

doi:10.1007/978-3-540-77587-4_212

Cited by 2 publications

(1 citation statement)

References 70 publications

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“…Fingerprinting approaches in microtiter plates, cocktail fingerprinting, and substrate microarrays were established to identify a specific enzyme of interest, to distinguish it from other closely related enzymes, and for diagnostic and quality control of enzyme‐containing samples 15–17. Although UV/vis‐ and fluorescence‐based assays are well developed, spectroscopic and spectrometric enantioselectivity assays were regularly established for a proof of concept investigating hydrolases, and in particular lipases 18, 19. Here, we report the use of high‐performance liquid chromatography (HPLC) in combination with circular dichroism (CD) detector screening for suitable alcohol dehydrogenases (ADH) to enantioselectively reduce 1‐phenyl‐2‐propyn‐3‐trimethylsilyl‐1‐on ( 1 ; Scheme ).…”

Section: Introductionmentioning

confidence: 99%

HPLC–CD selectivity assay for alcohol dehydrogenases

2011

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Enantioselective reductions are a key to successful target-oriented syntheses. Finding the most suitable conditions is often a tedious work that is especially hampered by the time-consuming analytical investigation. A possible solution is the combined use of high-performance liquid chromatography and circular dichroism to find a suitable system for providing enantiomerically pure alcohols. This investigation led to an efficient protocol for the alcohol dehydrogenase-catalyzed reduction of 1-phenyl-2-propyn-3-trimethylsilyl-1-on (1).

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Section: Introductionmentioning

confidence: 99%

HPLC–CD selectivity assay for alcohol dehydrogenases

2011

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Alternative hosts for functional (meta)genome analysis

Liebl

Angelov

Jüergensen

et al. 2014

Appl Microbiol Biotechnol

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Microorganisms are ubiquitous on earth, often forming complex microbial communities in numerous different habitats. Most of these organisms cannot be readily cultivated in the laboratory using standard media and growth conditions. However, it is possible to gain access to the vast genetic, enzymatic, and metabolic diversity present in these microbial communities using cultivation-independent approaches such as sequence- or function-based metagenomics. Function-based analysis is dependent on heterologous expression of metagenomic libraries in a genetically amenable cloning and expression host. To date, Escherichia coli is used in most cases; however, this has the drawback that many genes from heterologous genomes and complex metagenomes are expressed in E. coli either at very low levels or not at all. This review emphasizes the importance of establishing alternative microbial expression systems consisting of different genera and species as well as customized strains and vectors optimized for heterologous expression of membrane proteins, multigene clusters encoding protein complexes or entire metabolic pathways. The use of alternative host-vector systems will complement current metagenomic screening efforts and expand the yield of novel biocatalysts, metabolic pathways, and useful metabolites to be identified from environmental samples.

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Screening for Enantioselective Enzymes

Cited by 2 publications

References 70 publications

HPLC–CD selectivity assay for alcohol dehydrogenases

HPLC–CD selectivity assay for alcohol dehydrogenases

Alternative hosts for functional (meta)genome analysis

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