Enzyme selectivity as a tool in analytical chemistry
2007 (English)Licentiate thesis, comprehensive summary (Other scientific)
Enzymes are useful tools as specific analytical reagents. Two different analysis methods were developed for use in the separate fields of protein science and organic synthesis. Both methods rely on the substrate specificity of enzymes. Enzyme catalysis and substrate specificity is described and put in context with each of the two developed methods.
In paper I a method for C-terminal peptide sequencing was developed based on conventional Carboxypeptidase Y digestion combined with matrix assisted laser desorption/ionization mass spectrometry. An alternative nucleophile was used to obtain a stable peptide ladder and improve sequence coverage.
In paper II and III, three different enzymes were used for rapid analysis of enantiomeric excess and conversion of O-acylated cyanohydrins synthesized by a defined protocol. Horse liver alcohol dehydrogenase, Candida antarctica lipase B and pig liver esterase were sequentially added to a solution containing the O-acylated cyanohydrin. Each enzyme caused a drop in absorbance from oxidation of NADH to NAD+. The conversion and enantiomeric excess of the sample could be calculated from the relative differences in absorbance.
Place, publisher, year, edition, pages
Stockholm: KTH , 2007. , viii, 35 p.
Trita-BIO-Report, ISSN 1654-2312 ; 2007:5
substrate specificity, competing substrates, enantiomeric excess analysis, library screening, ladder sequencing, carboxypeptidase Y, 2-pyridylmethylamine, MALDI, Candia antarctica lipase B, pig liver esterase, horse liver alcohol dehydrogenase, acylated cyanohydrins
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:kth:diva-4377ISBN: 978-91-7178-674-6OAI: oai:DiVA.org:kth-4377DiVA: diva2:12022
2007-06-01, FB54, AlbaNova, Roslagstullsbacken 21, Stockholm, 10:00
Brumer, Harry, Docent
QC 201011082007-05-152007-05-152010-11-08Bibliographically approved
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