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Fermentation 2016, 2(1), 3; doi:10.3390/fermentation2010003

Characterizing the Phenotypic Responses of Escherichia coli to Multiple 4-Carbon Alcohols with Raman Spectroscopy

Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA 24061, USA
Author to whom correspondence should be addressed.
Academic Editor: George N. Bennett
Received: 3 December 2015 / Revised: 14 January 2016 / Accepted: 15 January 2016 / Published: 25 January 2016
(This article belongs to the Special Issue Metabolic Engineering)
View Full-Text   |   Download PDF [1821 KB, uploaded 25 January 2016]   |  


The phenotypic responses of E. coli cells exposed to 1.2% (v/v) of 1-butanol, 2-butanol, isobutanol, tert-butanol, and 1,4-butanediol were studied in near real-time using Raman spectroscopy. A method of “chemometric fingerprinting” was employed that uses multivariate statistics (principal component analysis and linear discriminant analysis) to identify E. coli phenotypic changes over a 180 min post-treatment time-course. A toxicity study showed extreme variability among the reduction in culture growth, with 1-butanol showing the greatest toxicity and 1,4-butanediol showing relatively no toxicity. Chemometric fingerprinting showed distinct phenotype clusters according to the type of alcohol: (i) 1-butanol and 2-butanol (straight chain alcohols); (ii) isobutanol and tert-butanol (branched chain alcohols); and (iii) control and 1,4-butanediol (no terminal alkyl end) treated cells. While the isobutanol and tert-butanol treated cells led to similar phenotypic responses, isobutanol was significantly more toxic. In addition, the phenotypic response was found to take place largely within 60 min of culture treatment; however, significant responses (especially for 1,4-butanediol) were still occurring at 180 min post-treatment. The methodology presented here identified different phenotypic responses to seemingly similar 4-carbon alcohols and can be used to study phenotypic responses of virtually any cell type under any set of environmental conditions or genetic manipulations. View Full-Text
Keywords: Raman spectroscopy; chemometric fingerprinting; linear discriminant analysis; alcohol toxicity Raman spectroscopy; chemometric fingerprinting; linear discriminant analysis; alcohol toxicity

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Zu, T.N.K.; Athamneh, A.I.M.; Senger, R.S. Characterizing the Phenotypic Responses of Escherichia coli to Multiple 4-Carbon Alcohols with Raman Spectroscopy. Fermentation 2016, 2, 3.

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