| Grant number: | 14/23362-8 |
| Support Opportunities: | Regular Research Grants |
| Duration: | March 01, 2015 - February 28, 2017 |
| Field of knowledge: | Physical Sciences and Mathematics - Chemistry - Organic Chemistry |
| Principal Investigator: | Márcio Santos da Silva |
| Grantee: | Márcio Santos da Silva |
| Host Institution: | Centro de Ciências Naturais e Humanas (CCNH). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil |
Abstract
Proteins are the most versatile among the various biological building blocks has lead to many industrial and biomedical applications. This scenario is mainly by recognized importance of chirality in biological processes, due to affinity differences for each enantiomer to receptors or enzymes. The design and screening of biocatalytic reactions are based on the same principles as traditional organic reactions (substrate, pH, solvent, temperature, etc.). In contrast, the informations obtained during the design and/or screening of biocatalytic processes are unclear by the inherent complexity of the enzyme. In addition to this, the methods for chiral discrimination, usually based on high performance liquid chromatography with chiral stationary phase (HPLC), are slow and can cause product decomposition, especially in more complex substrates. There are several methodologies for design and screening biocatalytic processes. Kinetic methods are arguably useful tools in the elucidation of reaction mechanisms. However, current methodologies and studies of enzyme-substrate interaction allow for practical, faster and more accurate results. These techniques provide efficient screening achieving a wide amount of information. Successful techniques common to most of experiments applied are mass spectrometry, infrared spectroscopy, electrophoresis and nuclear magnetic resonance - NMR. The project described here shows the development of protocols for design and chiral discrimination of biocatalytic reactions based on non-classical NMR methods. In the first part, will be developed a protocol to design the reaction of enzymatic kinetic resolution of amines by NMR (STD, T1, T2 and Diffusion) measuring the enzyme-substrate interactions. These techniques are distinguished by convenience, speed and different informations obtained, such as the interaction map of the substrate (substrate epitope), the type of enzyme-substrate interaction (the hydrogen interactions, electrostatic, aromatic interaction, among others) and which enantiomer will be formed. The protocol can be applied to verify the catalytic promiscuity of enzymes and inhibitory effect of substrates. The second step in this project is to use 125Te NMR for chiral discrimination by chiral solvation effect (ion pair) for the determination of enantiomeric excesses. The compounds containing Tellurium atom are synthesized from the reaction of enzymatic kinetic resolution of esters. For kinetic optimization of the reaction will be employed 1H NMR. The method reveals advantages as convenience, quickness, precision, no derivatization, recovery of the synthesized compounds and the possibility of working with unstable compounds, such as some alkyl tellurium compounds. (AU)
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