Success of microbial natural products discovery programs is dependent on quick and concise discrimination between isolates from diverse environments. However, laboratory isolation and identification of priority genera using current 16S rRNA PCR-based methods is both challenging and time-consuming. In clinical settings, MALDI-TOF MS is used to rapidly identify unknown bacterial pathogens, yet this has not been widely adapted for isolation of environmental bacteria. To expedite isolation of Gram-negative proteobacterial order Burkholderiales, we have created a main spectral (MSP) library for the Bruker Biotyper MALDI mass spectrometer that includes 95 entries of Burkholderia, Caballeronia, Paraburkholderia, and other environmentally related genera. We tested the effectiveness of the reference library by analyzing 49 unknown environmental isolate strains before comparing to results of traditional 16S sequencing. The MALDI Biotyper correctly identified all strains to the genus level when sufficient reference spectra were available. This dataset is freely available to the scientific research community for use in taxonomic assignments of environmentally-derived bacteria from the order Burkholderiales.

The Linington MSP Library (Burkholderia).btmsp dataset was acquired on the MALDI Biotyper microflex LT system (Bruker Daltonics). Over 2,250 individual mass spectrometry spectra were acquired in the range of 3,000 to 15,000 m/z to complete the 95 main spectra (MSP) entries. The acquired spectra were processed using associated manufacturer software (see publication for details). Library was exported and packaged in a *.btmsp format for use with MALDI Biotyper systems.

To load this dataset for use in discriminating environmental bacteria from environmental sampling campaigns, load into MALDI Biotyper systems as a supplemental library. Use MALDI Biotyper Automation Control software program to select “Linington MSP Library (Burkholderia)” in the wizard.