Microwave supported hydrolysis prepares Bacillus spores for proteomic analysis

Abstract

Rapid identification of Bacillus spores in the environment has depended primarily on a family of small acid soluble proteins (SASPs) as biomarkers. However, SASP sequences and molecular masses are similar or identical in some critical cases. For example, some strains of B. subtilis, and B. thuringiensis cannot be distinguished from strains of B. anthracis based on SASPs. Consequently, additional or alternative biomarkers should be sought. In this study microwaveassisted hot acid hydrolysis was coupled with mass spectrometry as a potentially powerful approach to the rapid automatable characterization of Bacillus spores. Hot acid provides lysis of the spores, Asp-selective hydrolysis of proteins, and peptides compatible with automated analysis of either peptide fingerprints or tandem mass spectra. Peptide biomarkers are compared here for a selection of Bacillus spores, and peptides unique to each spore type are identified.

MALDI-TOF Mass Spectrometric Detection of SARS-CoV-2 Using Cellulose Sulfate Ester Enrichment and Hot Acid Treatment

“…Hot acid protein hydrolysis occurs on both sides of aspartate, and the MASCOT program classifies some products as carrying missed cleavages. 26 , 27 We evaluated the effect of missed cleavages on the identification scores. The results show that the N protein is more confidently identified when missed cleavages are allowed ( Table 1 ).…”

“…Alternatively, the removal of PBS can be done by using C18 tips reported in our previous studies, which is more rapid and inexpensive. 26 , 27 After desalting, the sample can be directly applied to the following experimental procedure.…”

“…To remove 10×PBS from the virus solution, the sample was loaded into a 3K cutoff filter column and spun in a microcentrifuge at 14 000 g for 25 min. Alternatively, the removal of PBS can be done by using C18 tips reported in our previous studies, which is more rapid and inexpensive. , After desalting, the sample can be directly applied to the following experimental procedure.…”

Planning Implications Related to Sterilization-Sensitive Science Investigations Associated with Mars Sample Return (MSR)

“…Microwave-assisted hot acid extraction in HCl, formic acid, and acetic acid has also been used for spore lysis and extraction of acid-soluble proteins for the sequencing of small peptides using MALDI-MS techniques (Swatkoski et al, 2006;Chen et al, 2019).…”

Detection of Tuberculosis by The Analysis of Exhaled Breath Particles with High-resolution Mass Spectrometry

“…Therefore, there is a great need for the development of new screening tests that are operationally feasible in high-burdened settings and can rapidly and accurately detect active TB disease at a low cost-per-test. The promising application of mass spectrometry (MS) gains attention in the fields of environmental measurements, health care, and clinical diagnostics, etc [5][6][7] . Introduction of high-resolution orbitrap mass spectrometers (>100, 000 FWHM, full-width-at-half-maximum) enables the unambiguous assignment of chemical compositions without performing tandem mass spectrometry experiments 8,9 .…”