Rapid quantification of alcohol production in microorganisms based on nanostructure-initiator mass spectrometry (NIMS)

Kai Deng; Xi Wang; Nicole Ing; Paul Opgenorth; Markus de Raad; Jinho Kim; Blake A Simmons; Paul D Adams; Anup K Singh; Taek Soon Lee; Trent R Northen

doi:10.1016/j.ab.2022.114997

Rapid quantification of alcohol production in microorganisms based on nanostructure-initiator mass spectrometry (NIMS)

Anal Biochem. 2023 Feb 1:662:114997. doi: 10.1016/j.ab.2022.114997. Epub 2022 Nov 24.

Authors

Affiliations

¹ Joint BioEnergy Institute, Emeryville, CA, 94608, USA; Sandia National Laboratories, Livermore, CA, 94551, USA. Electronic address: kdeng@sandia.gov.
² Joint BioEnergy Institute, Emeryville, CA, 94608, USA; Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA.
³ Joint BioEnergy Institute, Emeryville, CA, 94608, USA; Sandia National Laboratories, Livermore, CA, 94551, USA.
⁴ Joint BioEnergy Institute, Emeryville, CA, 94608, USA; Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; University of California, Berkeley, CA, 94720, USA.
⁵ Joint BioEnergy Institute, Emeryville, CA, 94608, USA; Lawrence Livermore National Laboratory, Livermore, 94550, USA.
⁶ Joint BioEnergy Institute, Emeryville, CA, 94608, USA; Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA. Electronic address: trnorthen@lbl.gov.

PMID: 36435200
DOI: 10.1016/j.ab.2022.114997

Abstract

We described a mass spectrometry-based assay to rapidly quantify the production of primary alcohols directly from cell cultures. This novel assay used the combination of TEMPO-based oxidation chemistry and oxime ligation, followed by product analysis based on Nanostructure-Initiator Mass Spectrometry. This assay enables quantitative monitor both C5 to C18 alcohols as well as glucose and gluconate in the growth medium to support strain characterization and optimization. We find that this assay yields similar results to gas chromatography for isoprenol production but required much less acquisition time per sample. We applied this assay to gain new insights into P. Putida's utilization of alcohols and find that this strain largely could not grow on heptanol and octanol.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Ethanol
Gas Chromatography-Mass Spectrometry
Glucose
Mass Spectrometry / methods
Nanostructures* / chemistry

Substances

Glucose
Ethanol