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Surface enhanced Raman spectroscopy‐based evaluation of the membrane protein composition of the organohalide‐respiring Sulfurospirillum multivorans
Author(s) -
CiallaMay Dana,
Gadkari Jennifer,
Winterfeld Andreea,
Hübner Uwe,
Weber Karina,
Diekert Gabriele,
Schubert Torsten,
Goris Tobias,
Popp Jürgen
Publication year - 2021
Publication title -
journal of raman spectroscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.748
H-Index - 110
eISSN - 1097-4555
pISSN - 0377-0486
DOI - 10.1002/jrs.6029
Subject(s) - raman spectroscopy , chemistry , cofactor , electron acceptor , electron transport chain , surface enhanced raman spectroscopy , biochemistry , enzyme , raman scattering , physics , optics
Bacteria often employ different respiratory chains that comprise membrane proteins equipped with various cofactors. Monitoring the protein inventory that is present in the cells under a given cultivation condition is often difficult and time‐consuming. One example of a metabolically versatile bacterium is the microaerophilic organohalide‐respiring Sulfurospirillum multivorans . Here, we used surface enhanced Raman spectroscopy (SERS) to quickly identify the cofactors involved in the respiration of S. multivorans . We cultured the organism with either tetrachloroethene (perchloroethylene, PCE), fumarate, nitrate, or oxygen as electron acceptors. Because the corresponding terminal reductases of the four different respiratory chains harbor different cofactors, specific fingerprint signals in SERS were expected. Silver nanostructures fabricated by means of electron beam lithography were coated with the membrane fractions extracted from the four S. multivorans cultivations, and SERS spectra were recorded. In the case of S. multivorans cultivated with PCE, the recorded SERS spectra were dominated by Raman peaks specific for Vitamin B 12 . This is attributed to the high abundance of the PCE reductive dehalogenase (PceA), the key enzyme in PCE respiration. After cultivation with oxygen, fumarate, or nitrate, no Raman spectral features of B 12 were found.