Understanding the effects of culture conditions in bacterial growth: A biochemical perspective using Raman microscopy

Mukherjee, R and Verma, T and Nandi, D and Umapathy, S (2020) Understanding the effects of culture conditions in bacterial growth: A biochemical perspective using Raman microscopy. In: Journal of Biophotonics, 13 (1).

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Abstract

Rapid, sensitive and label-free methods to probe bacterial growth irrespective of the culture conditions can shed light on the mechanisms by which bacteria adapt to different environmental stimuli. Raman spectroscopy can rapidly and continuously monitor the growth of bacteria under varied conditions. In this study, the growth of Escherichia coli in Luria broth (nutrient rich conditions) and minimal media with either glucose or glycerol as carbon source (nutrient limiting conditions) is profiled using Raman spectroscopy. Moreover, the study also gives insights into the altered bacterial biochemistry upon exposure to low- (25°C) and high-temperature (45°C) stress. Raman spectral measurement was performed on bulk bacteria cultured under laboratory conditions. A detailed analysis of the spectra as a function of bacterial growth reveals changes in Raman band intensities/area of biomolecules such as DNA, proteins and lipids. We also report five novel ratiometric markers (I830/I810, I1126/I1100, I1340/I1440, I1207/I1240 and I1580/I1440) that can identify the phase of growth, independent of the culture condition. Unsupervised multivariate methods like Principal Component Analysis also corroborate the aforementioned markers of growth. Altogether, our findings highlight the potential of Raman spectroscopy in yielding universal biochemical signatures that may be indicative of stress and aging in a growth milieu. © 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Item Type:	Journal Article
Publication:	Journal of Biophotonics
Publisher:	Wiley-VCH Verlag
Additional Information:	The copyright for this article belongs to Wiley-VCH Verlag.
Keywords:	Biomolecules; Escherichia coli; Multivariant analysis; Nutrients; Principal component analysis; Raman spectroscopy, Bacterial growth; Environmental stimuli; Laboratory conditions; Multi variate analysis; Multivariate methods; Raman micro spectroscopy; Raman spectral measurements; Ratiometric analysis, Spectrum analysis, bacterium; Escherichia coli; microscopy; principal component analysis; Raman spectrometry, Bacteria; Escherichia coli; Microscopy; Principal Component Analysis; Spectrum Analysis, Raman
Department/Centre:	Division of Biological Sciences > Biochemistry Division of Chemical Sciences > Inorganic & Physical Chemistry Division of Interdisciplinary Sciences > Centre for Biosystems Science and Engineering Division of Physical & Mathematical Sciences > Instrumentation Appiled Physics
Date Deposited:	08 Feb 2023 09:18
Last Modified:	08 Feb 2023 09:18
URI:	https://eprints.iisc.ac.in/id/eprint/80075

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