Abstract
This study shows the application of the Fourier transform mid-infrared spectroscopy (FT-MIR) associated with high-throughput technology to study the biochemical fingerprints of different Saccharomyces cerevisiae strains transformed with the same expression system along the similar cultivation in bioreactor. The phenotype, as well as the cellular metabolism and recombinant cyprosin biosynthesis, were determined. The differences observed were confirmed by conventional cyprosin activity protocol, and the metabolic evolution was analyzed using high-performance liquid chromatography technique. The spectral analysis based on chemometrics tools, such as the principal component analysis, is a useful methodology for the phenotypes characterization as well as the specific metabolic states along the cultivations according to the clusters created. The ratio bands of spectra also represented a useful tool to evaluate the metabolic and biochemical differences between both expression systems, allowing to have an additional parameter to the biomolecular comparison. Therefore, high-throughput FT-MIR spectroscopy associated with multivariate data analysis represent a valuable strategy for extracting significant specific biomolecular information along the cultivation, providing a complete bioprocess analysis, once it detects slight molecular changes which it will be useful for screening and optimization process in the biotechnological or pharmaceutical industry.
Original language | English |
---|---|
Pages (from-to) | 1-10 |
Number of pages | 10 |
Journal | Journal of Biotechnology |
Volume | 260 |
DOIs | |
Publication status | Published - 20 Oct 2017 |
Bibliographical note
Publisher Copyright:© 2017 Elsevier B.V.
Funding
We thank Dr. Filomena Calixto for providing S. cerevisiae BJ1991and W303-1A strains transformed with CYPRO11 gene. We also thank Professor António Mendonça from the University of Beira Interior by the availability of laboratory equipment and facilities related to spectra acquisition. The present work was partially conducted in the Health & Engineering Laboratory resulting from the Protocol between the Instituto Politécnico de Lisboa and the Universidade Católica Portuguesa. This work was supported by CLARO research project funded by Agência de Inovação -ADI agency (Portugal).
Funders | Funder number |
---|---|
CLARO | |
Agência Nacional de Inovação |
Keywords
- FT-MIR spectroscopy
- High-throughput analysis
- Principal components analysis
- Recombinant cyprosin B
- Screening process