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Published on: February 19, 2016
Detection of Penicillin G Produced by Penicillium chrysogenum with Raman Microspectroscopy and Multivariate Curve
Shumpei Horii1,2, Masahiro Ando3,4, Ashok Zachariah Samuel3
1Department of Advanced Science Engineering, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo 169-8555, Japan.
Abstract:
Raman microspectroscopy is a minimally invasive technique that can identify molecules without labeling. In this study, we demonstrate the detection of penicillin G inside Penicillium chrysogenum KF425 fungal cells. Raman spectra acquired from the fungal cells had highly overlapped spectroscopic signatures and hence were analyzed with multivariate curve resolution by alternating least-squares (MCR-ALS) to extract the spectra of individual molecular constituents. In addition to detecting spatial distribution of multiple constituents such as proteins and lipids inside the fungal body, we could also observe the subcellular localization of penicillin G. This methodology has the potential to be employed in screening the production of bioactive compounds by microorganisms.
Insights
Raman microspectroscopy detected penicillin G within fungal cells. This technique, using multivariate analysis, maps molecular distribution and aids in screening bioactive compound production by microorganisms.
Area of Science:
- Biotechnology
- Microbiology
- Spectroscopy
Background:
- Raman microspectroscopy offers label-free molecular identification.
- Analyzing complex biological samples requires advanced data processing.
- Understanding intracellular compound localization is crucial for optimizing production.
Purpose of the Study:
- To demonstrate the detection and subcellular localization of penicillin G in *Penicillium chrysogenum* KF425 using Raman microspectroscopy.
- To apply multivariate curve resolution by alternating least-squares (MCR-ALS) for analyzing complex spectral data from fungal cells.
- To explore the potential of this methodology for screening microbial bioactive compound production.
Main Methods:
- Raman microspectroscopy was employed for non-invasive molecular analysis.
- Multivariate curve resolution by alternating least-squares (MCR-ALS) was used to resolve overlapping spectral signatures.
- Spatial distribution of molecules, including penicillin G, proteins, and lipids, was mapped within fungal cells.
Main Results:
- Penicillin G was successfully detected inside *Penicillium chrysogenum* KF425 fungal cells.
- The subcellular localization of penicillin G was observed.
- Spatial distribution of proteins and lipids within the fungal cells was also determined.
- MCR-ALS effectively extracted individual molecular spectra from complex cellular environments.
Conclusions:
- Raman microspectroscopy combined with MCR-ALS is a powerful tool for analyzing molecular composition and distribution in microbial cells.
- This approach enables the visualization of bioactive compound localization at the subcellular level.
- The methodology holds significant potential for high-throughput screening of microorganisms for enhanced production of valuable compounds.
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