Bioprofiling of unknown antibiotics in herbal extracts: Development of a streamlined direct bioautography using Bacillus subtilis linked to mass spectrometry

Jamshidi-Aidji, Maryam; Morlock, Gertrud E.

doi:10.1016/j.chroma.2015.09.061

Cited by 65 publications

(35 citation statements)

References 29 publications

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“…However, the use of enzyme inhibition assays by the Ellman method is the first obligatory step in determining the inhibition of AChE and BChE of the total extracts (Ellman et al, 1961). Of note, the Ellman method is accompanied by bioautographs with thin layer chromatography (TLC) (Rhee et al, 2001;Marston et al, 2002) and a consequent densitometry (Mroczek and Mazurek, 2009;Jamshidi-Aidji and Morlock, 2015) provides a simple, effective approach to postulate fractions or alkaloids with inhibitory activity of cholinesterases. In this regard, it is generally assumed that the active site of mammalian AChE, for example, is identical in all tissues at all ages (Herkert et al, 2010;Jamshidi-Aidji and Morlock, 2015).…”

Section: Introductionmentioning

confidence: 99%

“…Of note, the Ellman method is accompanied by bioautographs with thin layer chromatography (TLC) (Rhee et al, 2001;Marston et al, 2002) and a consequent densitometry (Mroczek and Mazurek, 2009;Jamshidi-Aidji and Morlock, 2015) provides a simple, effective approach to postulate fractions or alkaloids with inhibitory activity of cholinesterases. In this regard, it is generally assumed that the active site of mammalian AChE, for example, is identical in all tissues at all ages (Herkert et al, 2010;Jamshidi-Aidji and Morlock, 2015). However, kinetic in vitro studies of human and animal AChE have shown substantial species differences in the enzymatic potency (Luo et al, 2007;Jamshidi-Aidji and Morlock, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Alkaloids of Amaryllidaceae as Inhibitors of Cholinesterases (AChEs and BChEs): An Integrated Bioguided Study

Cortés

Sierra

Álzate

et al. 2017

Phytochemical Analysis

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Alkaloids of Amaryllidaceae as Inhibitors of Cholinesterases (AChEs and BChEs): An Integrated Bioguided Study

Cortés

Sierra

Álzate

et al. 2017

Phytochemical Analysis

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“…20 TLC-bioautography allows the localization of the active principles during the separation process of even complex extracts directly on the TLC plate. Although bioautography appears to be increasingly utilized to detect antimicrobial activities, 21,22 there are no reports of its use with Mtb .…”

Section: Resultsmentioning

confidence: 99%

Bioautography with TLC-MS/NMR for Rapid Discovery of Anti-tuberculosis Lead Compounds from Natural Sources

et al. 2016

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While natural products constitute an established source of lead compounds, the classical iterative bioassay-guided isolation process is both time- and labor-intensive and prone to failing to identify active minor constituents. (HP)TLC-bioautography-MS/NMR, which combines cutting-edge microbiological, chromatographic, and spectrometric technologies, was developed to accelerate anti-tuberculosis (TB) drug discovery from natural sources by acquiring structural information at a very early stage of the isolation process. Using the avirulent, bioluminescent Mtb strain mc27000 luxABCDE, three variations of bioautography were evaluated and optimized for sensitivity in detecting anti-TB agents, including established clinical agents and new leads with novel mechanisms of action. Several exemplary applications of this approach to microbial extracts demonstrate its potential as a routine method in anti-TB drug discovery from natural sources.

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“…Thin layer chromatography‐direct bioautography (TLC‐DB) overcomes these limitations by applying a bacterial suspension directly onto the TLC plate . ZoIs of bacterial growth are visualised using metabolic stains, typically tetrazolium salts such as 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) or 2‐(4‐iodophenyl)‐3‐(4‐nitrophenyl)‐5‐phenyl‐2 H ‐tetrazolium chloride (INT) . These salts are reduced by dehydrogenases in viable cells to produce purple formazan, thus ZoIs are visualised as white spots on a purple background .…”

Section: Introductionmentioning

confidence: 99%

“…19 ZoIs of bacterial growth are visualised using metabolic stains, typically tetrazolium salts such as 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) or 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride (INT). [20][21][22][23][24] These salts are reduced by dehydrogenases in viable cells to produce purple formazan, thus ZoIs are visualised as white spots on a purple background. 24,25 Lead compounds are then extracted and characterised.…”

Section: Introductionmentioning

confidence: 99%

Characterisation and screening of antimicrobial essential oil components against clinically important antibiotic‐resistant bacteria using thin layer chromatography‐direct bioautography hyphenated with GC‐MS, LC‐MS and NMR

Owen

White

Laird

2018

Phytochemical Analysis

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Introduction The antimicrobial activity of many essential oils (EOs) is well established, indicating that EOs may be a source of compounds for antimicrobial drug development. Thin layer chromatography‐direct bioautography (TLC‐DB) can quickly identify antimicrobial components in complex mixtures and can be applied to the screening of EOs for lead compounds. Objectives This study aimed to identify antimicrobial components of oregano, rosewood and cumin EOs against antibiotic‐sensitive and ‐resistant bacteria using TLC‐DB and a multi‐faceted approach of GC‐MS, LC‐MS and NMR techniques to characterise bioactive compounds. The study also aimed to quantify the antimicrobial activity of bioactive compounds in order to evaluate their potential for the development of therapies against antibiotic‐resistant bacteria. Materials and Methods EOs were eluted on TLC plates and sprayed with a suspension of Staphylococcus aureus, Enterococcus faecium, Escherichia coli or Pseudomonas aeruginosa (antibiotic‐sensitive and ‐resistant isolates). Zones of inhibition, visualised with iodonitrotetrazolium chloride, were subject to GC‐MS, LC‐MS and NMR to characterise the bioactive compounds. Results Seven compounds were identified from the three EOs using GC‐MS, while LC‐MS and NMR failed to detect the presence of any further non‐volatile or heat labile compounds. Carvacrol was most antimicrobial compound identified, with minimum inhibitory concentrations ranging 0.99–31.62 mM. Conclusion The identified antimicrobial compounds present in oregano, rosewood and cumin EOs including carvacrol may be candidates for the development of novel antimicrobial therapies against antibiotic‐resistant bacteria.

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Bioprofiling of unknown antibiotics in herbal extracts: Development of a streamlined direct bioautography using Bacillus subtilis linked to mass spectrometry

Cited by 65 publications

References 29 publications

Alkaloids of Amaryllidaceae as Inhibitors of Cholinesterases (AChEs and BChEs): An Integrated Bioguided Study

Alkaloids of Amaryllidaceae as Inhibitors of Cholinesterases (AChEs and BChEs): An Integrated Bioguided Study

Bioautography with TLC-MS/NMR for Rapid Discovery of Anti-tuberculosis Lead Compounds from Natural Sources

Characterisation and screening of antimicrobial essential oil components against clinically important antibiotic‐resistant bacteria using thin layer chromatography‐direct bioautography hyphenated with GC‐MS, LC‐MS and NMR

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