Detection of Bacteriocins by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Rose, Natisha L.; Sporns, Peter; McMullen, Lynn M.

doi:10.1128/aem.65.5.2238-2242.1999

Cited by 31 publications

(13 citation statements)

References 30 publications

(54 reference statements)

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“…Published linear mode MALDI-TOF spectra were of very low resolution, and the variance in the determined mass numbers made unambiguous assignments impossible. 7,35,36 The methodology introduced in this communication can be generally applied for efficient screening of bacteria to identify novel bacteriocins and lantibiotics. The sample preparation -whole cells embedded into the DHBs-matrix crystals -exhibited a remarkable stability, which demonstrates the robustness of the technique.…”

Section: Discussionmentioning

confidence: 99%

Whole‐cell matrix‐assisted laser desorption/ionization mass spectrometry for rapid identification of bacteriocin/lantibiotic‐producing bacteria

Stein

2008

Rapid Comm Mass Spectrometry

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The potential of whole-cell matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) for the discovery of microorganisms that produce lantibiotics and/or bacteriocins in the mass range 3-5 kDa is demonstrated. Cells of the antibiotic-producing model strain Bacillus subtilis ATCC 6633 were grown on Landy agar plates, and prepared with a 9:1 mixture of 2,5-dihydroxybenzoic acid and 2-hydroxy-5-methoxybenzoic acid (DHBs) as matrix using the bottom-layer method. Time-course analyses showed that cells grown for 1-2 days on agar plates gave the best results in terms of intensity of the bacteriocin/lantibiotic signals. Even after storage of matrix-crystal-embedded cells for several days high-resolution spectra (>10,000) could be obtained. To verify the applicability of this technique, different Bacillus wild-type and gene deletion mutant cells were analyzed. In most of the cases the known antibiotic phenotype of the producer could be corroborated by whole-cell MALDI-TOFMS, particularly for the lantibiotics subtilin and ericin. The preparation recipe for whole-cell analysis by MALDI-TOF was also highly sensitive for hydrophobic lipopeptide antibiotics like surfactin, mycosubtilin, fengycin, and bacillomycin in the lower mass range (1-1.5 kDa).

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Section: Discussionmentioning

confidence: 99%

Whole‐cell matrix‐assisted laser desorption/ionization mass spectrometry for rapid identification of bacteriocin/lantibiotic‐producing bacteria

Stein

2008

Rapid Comm Mass Spectrometry

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“…Samples were prepared for MALDI-TOF MS analysis by the method described by Rose et al (1999). Samples for analysis included initial and residual nisin solutions, extracts of raw and cooked meat soaked in nisin, and raw and cooked juice with nisin added.…”

Section: Maldi-tof Msmentioning

confidence: 99%

Inactivation of Nisin by Glutathione in Fresh Meat

Rose

Sporns

Stiles

et al. 1999

Journal of Food Science

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Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to determine the fate of nisin in meat products. Nisin in 0.02N HCl was added to fresh and cooked meat and meat juice and samples were stored at 4 °C overnight. Residual nisin solutions and meat juice and meat extract supernatants were analyzed for antimicrobial activity and for nisin. Nisin was recovered from cooked meat extract and cooked meat juice; however, only nisin bound to a food component was detected in fresh meat extract. Mass spectra for raw meat and juice showed a signal 307 Da greater than the mass of nisin. Results indicated that nisin was likely inactivated in raw meat by an enzymatic reaction with glutathione.

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“…However, bacterial culture media are often highly complex due to their content of salts, nitrogen and carbon sources, and small hydrophilic metabolites. MS analyses of peptide antibiotics within such media was possible after prior butanolic extraction in order to enrich and desalt the analytes prior to measurements,8,, 9 or after washing of membrane spotted samples 10. However, direct MS monitoring of the culture media without any time‐consuming sample preparations would be preferred for both, analytical and preparative purposes.…”

mentioning

confidence: 99%

Maturation of the lantibiotic subtilin: matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry to monitor precursors and their proteolytic processing in crude bacterial cultures

Stein

Entian

2001

Rapid Comm Mass Spectrometry

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Bacillus subtilis synthesizes the lanthionine containing 32-amino-acid peptide antibiotic (lanti-biotic) subtilin from a ribosomally generated 56-amino-acid precursor pre-propeptide by extensive posttranslational modifications. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was used to monitor the production of matured subtilin within crude samples taken from B. subtilis culture media without prior fractionation. The processing reaction of subtilin was blocked with the serine protease inhibitor phenylmethylsulfonyl fluoride and different subtilin precursor peptides in the molecular mass range up to 6220 were observed. Two of these species were isolated by reversed-phase high-performance liquid chromatography (HPLC) and structurally analyzed by post-source decay MALDI-TOFMS. We provide evidence that the precursor species comprise the posttranslational modified C-terminal part of subtilin to which leader peptide moieties with different chain lengths are attached. These antimicrobial-inactive species could be processed to antibiotic-active subtilin by incubation with culture media of different subtilin-nonproducing B. subtilis strains as indicated by a combination of antimicrobial growth assays and MALDI-TOFMS analyses. These achievements are strong evidence for the sensitivity of MALDI-TOFMS methodology that allows straightforward investigations of analytes even in complex mixtures without time-consuming sample preparations.

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Detection of Bacteriocins by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Cited by 31 publications

References 30 publications

Whole‐cell matrix‐assisted laser desorption/ionization mass spectrometry for rapid identification of bacteriocin/lantibiotic‐producing bacteria

Whole‐cell matrix‐assisted laser desorption/ionization mass spectrometry for rapid identification of bacteriocin/lantibiotic‐producing bacteria

Inactivation of Nisin by Glutathione in Fresh Meat

Maturation of the lantibiotic subtilin: matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry to monitor precursors and their proteolytic processing in crude bacterial cultures

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