Ultrasonic enzyme assays. Real time monitoring of enzyme reactions in solutions and complex media using high-resolution ultrasonic spectroscopy
✍ Scribed by V. Buckin; P. Resa; E. Kudryashov
- Publisher
- Elsevier
- Year
- 2009
- Tongue
- English
- Weight
- 83 KB
- Volume
- 25
- Category
- Article
- ISSN
- 1871-6784
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✦ Synopsis
oligomers -between 5000 and 1000 Da, and to assess the possibility of a more controlled depolymerisation, by improving the polydispersity index (PDI) from an acid and alkaline hydrolysis on previous studies.
We worked with three kinds of enzymes, two of them commercially available -lipases and esterases, and with a specific PHB depolymerase (His-tagged phaZ).
Through analysing the activity of each kind of enzyme, we measure its efficiency to produce specific low-molecular-weight oligomers.
Depolymerisations were tested under different conditions, changing the exposure time and concentration for each used enzyme. Depolymerisation monitor and products were analysed with Gel Permeation Chromatography (GPC), Fourier Transform Infrared Spectroscopy (FTIR), Differential Scanning Calorimetry (DSC) and Nuclear Magnetic Resonance Spectroscopy (NMR).
We suggest that using enzymes for a controlled depolymerisation process will allow us to obtain oligomers of a desired low-molecular-weight, to the extent that we can determine a generic enzyme structure that brings us the best results.