Rapid and improved assay of surfactins from Bacillus subtilis, 203R via UPLC-ESI-MS

UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
David M. Wright (Creator)
Institution
The University of North Carolina at Greensboro (UNCG )
Web Site: http://library.uncg.edu/
Advisor
Nadja Cech

Abstract: To better bridge research with commercialization, this project sought to develop an improved analytical method for the assay of biosurfactants known as surfactins from Bacillus subtilis. We sought to compare levels of production from various strains of B. subtilis, including strain 203R, which was isolated from areas prone to oil degraders. The use of Ultra-Performance Liquid Chromatography coupled with High Resolution Mass Spectrometry proved beneficial for the selective identification and quantification of surfactin lipopeptides. Surfactins were eluted in under 10 minutes and quantified using the total area of generated ions. Near baseline separation across the envelope gave an added layer of identification to the ions responsible for surfactins. These lipopeptides, having similar molecular masses to the iturins, nearly coelute complicating the analysis. By coupling retention time with fragmentation pattern, some isoforms could be distinguished. Validation of the method was achieved by obtaining 4 calibration curves on different days and applying linear regression analysis. Strain 203R was shown to be a superior producer of surfactins than the previously reported model strain ATCC 21332.

Additional Information

Publication
Thesis
Language: English
Date: 2018
Keywords
Bacillus subtilis, Linear Dynamic Range, Lipopeptide, Residuals, Surfactin
Subjects
Bacillus subtilis
Biosurfactants
Microbiological assay

Email this document to