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Enzyme selectivity as a tool in analytical chemistry

<p>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.</p><p>In <strong>paper I </strong>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.</p><p>In paper<strong> II </strong>and <strong>III</strong>, 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,<em> Candida antarctica</em> lipase<strong> </strong>B<strong> </strong>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<sup>+</sup>. The conversion and enantiomeric excess of the sample could be calculated from the relative differences in absorbance.</p>

Identiferoai:union.ndltd.org:UPSALLA/oai:DiVA.org:kth-4377
Date January 2007
CreatorsHamberg, Anders
PublisherKTH, School of Biotechnology (BIO), Stockholm : Bioteknologi
Source SetsDiVA Archive at Upsalla University
LanguageEnglish
Detected LanguageEnglish
TypeLicentiate thesis, comprehensive summary, text
RelationTrita-BIO-Report, 1654-2312 ; 2007:5

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