The plant alkaloid piperine as a potential inhibitor of ethidium bromide efflux in Mycobacterium smegmatis

2011 ◽  
Vol 60 (2) ◽  
pp. 223-229 ◽  
Author(s):  
Jing Jin ◽  
Jiyu Zhang ◽  
Na Guo ◽  
Haihua Feng ◽  
Lei Li ◽  
...  
Keyword(s):  
Ethidium Bromide ◽  
Mycobacterium Smegmatis ◽  
Efflux Pump ◽  
Black Pepper ◽  
Piper Nigrum ◽  
Fluorometric Method ◽  
Piper Longum ◽  
Carbonyl Cyanide ◽  
Inhibition Ability ◽  
Positive Controls

Piperine, a major plant alkaloid found in black pepper (Piper nigrum) and long pepper (Piper longum), has shown potential for inhibiting the efflux pump (EP) of Staphylococcus aureus. In this study, a modulation assay showed that piperine could decrease the MIC of ethidium bromide (EtBr) twofold at 32 μg ml−1 and fourfold at 64 μg ml−1 against Mycobacterium smegmatis mc2 155 ATCC 700084. A real-time, 96-well plate fluorometric method was employed to evaluate the EP inhibition ability of piperine in M. smegmatis. Reserpine, chlorpromazine, verapamil and carbonyl cyanide m-chlorophenylhydrazone were used as positive controls. Piperine significantly enhanced accumulation and decreased the efflux of EtBr in M. smegmatis, which suggests that it has the ability to inhibit mycobacterial EPs.

scholarly journals Efflux Pump-Mediated Intrinsic Drug Resistance in Mycobacterium smegmatis

2004 ◽  
Vol 48 (7) ◽  
pp. 2415-2423 ◽  
Author(s):  
Xian-Zhi Li ◽  
Li Zhang ◽  
Hiroshi Nikaido
Keyword(s):  
Drug Resistance ◽  
Ethidium Bromide ◽  
Mycobacterium Smegmatis ◽  
Efflux Pump ◽  
Upstream Region ◽  
Drug Efflux ◽  
Intact Cells ◽  
Intrinsic Drug Resistance ◽  
Multiple Drugs ◽  
Increased Susceptibility

ABSTRACT The Mycobacterium smegmatis genome contains many genes encoding putative drug efflux pumps. Yet with the exception of lfrA, it is not clear whether these genes contribute to the intrinsic drug resistance of this organism. We showed first by reverse transcription (RT)-PCR that several of these genes, including lfrA as well as the homologues of Mycobacterium tuberculosis Rv1145, Rv1146, Rv1877, Rv2846c (efpA), and Rv3065 (mmr and emrE), were expressed at detectable levels in the strain mc2155. Null mutants each carrying an in-frame deletion of these genes were then constructed in M. smegmatis. The deletions of the lfrA gene or mmr homologue rendered the mutant more susceptible to multiple drugs such as fluoroquinolones, ethidium bromide, and acriflavine (two- to eightfold decrease in MICs). The deletion of the efpA homologue also produced increased susceptibility to these agents but unexpectedly also resulted in decreased susceptibility to rifamycins, isoniazid, and chloramphenicol (two- to fourfold increase in MICs). Deletion of the Rv1877 homologue produced some increased susceptibility to ethidium bromide, acriflavine, and erythromycin. The upstream region of lfrA contained a gene encoding a putative TetR family transcriptional repressor, dubbed LfrR. The deletion of lfrR elevated the expression of lfrA and produced higher resistance to multiple drugs. Multidrug-resistant single-step mutants, independent of LfrA and attributed to a yet-unidentified drug efflux pump (here called LfrX), were selected in vitro and showed decreased accumulation of norfloxacin, ethidium bromide, and acriflavine in intact cells. Finally, use of isogenic β-lactamase-deficient strains showed the contribution of LfrA and LfrX to resistance to certain β-lactams in M. smegmatis.

scholarly journals mmpL7 Gene of Mycobacterium tuberculosis Is Responsible for Isoniazid Efflux in Mycobacterium smegmatis

2005 ◽  
Vol 49 (11) ◽  
pp. 4775-4777 ◽  
Author(s):  
Maria R. Pasca ◽  
Paola Guglierame ◽  
Edda De Rossi ◽  
Francesca Zara ◽  
Giovanna Riccardi
Keyword(s):  
Mycobacterium Tuberculosis ◽  
High Resistance ◽  
Mycobacterium Smegmatis ◽  
Efflux Pump ◽  
Gene Encoding ◽  
Resistance Level ◽  
Carbonyl Cyanide ◽  
Resistance Nodulation Division ◽  
Efflux Pump Inhibitors ◽  
Energy Dependent

ABSTRACT The Mycobacterium tuberculosis mmpL7 gene, encoding a hypothetical resistance nodulation division transporter, confers a high resistance level to isoniazid when overexpressed in Mycobacterium smegmatis. The resistance level decreased in the presence of the efflux pump inhibitors reserpine and CCCP (carbonyl cyanide m-chlorophenylhydrazone). Energy-dependent efflux of isoniazid from M. smegmatis cells expressing the mmpL7 gene was observed.

scholarly journals Characterization of P55, a Multidrug Efflux Pump inMycobacterium bovis and Mycobacterium tuberculosis

2001 ◽  
Vol 45 (3) ◽  
pp. 800-804 ◽  
Author(s):  
Pedro E. A. Silva ◽  
Fabiana Bigi ◽  
Marı́a de la Paz Santangelo ◽  
Maria Isabel Romano ◽  
Carlos Martı́n ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mycobacterium Smegmatis ◽  
Efflux Pump ◽  
Tetracycline Resistance ◽  
Drug Efflux ◽  
Multidrug Efflux ◽  
Multidrug Efflux Pump ◽  
Reading Frame ◽  
Carbonyl Cyanide

ABSTRACT The Mycobacterium bovis P55 gene, located downstream from the gene that encodes the immunogenic lipoprotein P27, has been characterized. The gene was identical to the open reading frame of the Rv1410c gene in the genome of Mycobacterium tuberculosisH37Rv, annotated as a probable drug efflux protein. Genes similar toP55 were present in all species of the M. tuberculosis complex and other mycobacteria such asMycobacterium leprae and Mycobacterium avium. By Western blotting, P55 was located in the membrane fraction ofM. bovis. When transformed into Mycobacterium smegmatis after cloning, P55 conferred aminoglycoside and tetracycline resistance. The levels of resistance to streptomycin and tetracycline conferred by P55 were decreased in the presence of the protonophore carbonyl cyanide m-chlorophenylhydrazone and the pump inhibitors verapamil and reserpine. M. smegmatiscells expressing the plasmid-encoded P55 accumulated less tetracycline than the control cells. We conclude that P55 is a membrane protein implicated in aminoglycoside and tetracycline efflux in mycobacteria.

scholarly journals Rv2686c-Rv2687c-Rv2688c, an ABC Fluoroquinolone Efflux Pump in Mycobacterium tuberculosis

2004 ◽  
Vol 48 (8) ◽  
pp. 3175-3178 ◽  
Author(s):  
Maria Rosalia Pasca ◽  
Paola Guglierame ◽  
Fabio Arcesi ◽  
Marco Bellinzoni ◽  
Edda De Rossi ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Abc Transporter ◽  
Mycobacterium Smegmatis ◽  
Efflux Pump ◽  
Resistance Level ◽  
Carbonyl Cyanide ◽  
Efflux Pump Inhibitors ◽  
Energy Dependent

ABSTRACT The Mycobacterium tuberculosis Rv2686c-Rv2687c-Rv2688c operon, encoding an ABC transporter, conferred resistance to ciprofloxacin and, to a lesser extent, norfloxacin, moxifloxacin, and sparfloxacin to Mycobacterium smegmatis. The resistance level decreased in the presence of the efflux pump inhibitors reserpine, carbonyl cyanide m-chlorophenylhydrazone, and verapamil. Energy-dependent efflux of ciprofloxacin from M. smegmatis cells containing the Rv2686c-Rv2687c-Rv2688c operon was observed.

Antileishmanial Alkaloids from Black Pepper (Piper nigrum)

Planta Medica ◽  
2011 ◽  
Vol 77 (05) ◽  
Author(s):  
HRW Dharmaratne ◽  
BL Tekwani ◽  
NPD Nanayakkara
Keyword(s):  
Black Pepper ◽  
Piper Nigrum

DETERMINATION OF THE EFFICIENCY AND EFFICACY OF BIO-CHEMICAL FERTILIZER ON THE SOIL, YIELD AND GROWTH PERFORMANCE OF BLACK PEPPER (Piper nigrum L.)

2015 ◽  
Vol 4 (3) ◽  
pp. 460-468
Author(s):  
Yap Chin Ann
Keyword(s):  
Growth Performance ◽  
Nutrient Management ◽  
High Growth ◽  
Black Pepper ◽  
Chemical Fertilizer ◽  
Piper Nigrum ◽  
High Fertility ◽  
Fertility Level ◽  
Area Index ◽  
K 12

The last nutrient management review of black pepper was done in 1968. There is, therefore, a need to develop new technology to improve pepper production and transfer that technology to production site. This experiment was carried out to study the effect of newly developed biochemical fertilizer on some physiological characteristics, yield and soil fertility of pepper. The treatment consisted of T1 (BS): chemical fertilizer (N:12%, P:12%, K:17%); T2 (BK1): biochemical fertilizer F1 N:15%, P:5%, K:14) and T3 (BK2): biochemical fertilizer F2 (N:13%, P:4%, K:12). The biochemical fertilizer F1 out-yielded chemical and biochemical fertilizer F2 by 75.38% and 16.45% respectively with the higher yield being associated with various phonotypical alterations, which are reported here. Significant measureable changes were observed in physiological processes and plant characteristics, such as large leaf area index, more chlorophyll content and high photosynthesis rate coupled with lower transpiration rate in biochemical fertilizer F1(BK1) treatment compared with other treatment. The high fertility level in biochemical fertilizer F1 and biochemical fertilizer F2 (BK2) reflected the important of organic material in improving soil quality. In conclusion, the achieve high growth performance and yield in pepper, chemical fertilizer alone is insufficient whilst combination of organic and inorganic fertilizer with balance nutrient content gave a significant increase in yield and growth of pepper. 

scholarly journals A Simple Method for Assessment of MDR Bacteria for Over-Expressed Efflux Pumps

2013 ◽  
Vol 7 (1) ◽  
pp. 72-82 ◽  
Author(s):  
Marta Martins ◽  
Matthew P McCusker ◽  
Miguel Viveiros ◽  
Isabel Couto ◽  
Séamus Fanning ◽  
...  
Keyword(s):  
Ethidium Bromide ◽  
Efflux Pump ◽  
Efflux Pumps ◽  
Outer Cell ◽  
Gram Negative Bacteria ◽  
Drug Resistant ◽  
Topoisomerase Iv ◽  
Simple Method ◽  
Gram Negative ◽  
Over Expression

It is known that bacteria showing a multi-drug resistance phenotype use several mechanisms to overcome the action of antibiotics. As a result, this phenotype can be a result of several mechanisms or a combination of thereof. The main mechanisms of antibiotic resistance are: mutations in target genes (such as DNA gyrase and topoisomerase IV); over-expression of efflux pumps; changes in the cell envelope; down regulation of membrane porins, and modified lipopolysaccharide component of the outer cell membrane (in the case of Gram-negative bacteria). In addition, adaptation to the environment, such as quorum sensing and biofilm formation can also contribute to bacterial persistence. Due to the rapid emergence and spread of bacterial isolates showing resistance to several classes of antibiotics, methods that can rapidly and efficiently identify isolates whose resistance is due to active efflux have been developed. However, there is still a need for faster and more accurate methodologies. Conventional methods that evaluate bacterial efflux pump activity in liquid systems are available. However, these methods usually use common efflux pump substrates, such as ethidium bromide or radioactive antibiotics and therefore, require specialized instrumentation, which is not available in all laboratories. In this review, we will report the results obtained with the Ethidium Bromide-agar Cartwheel method. This is an easy, instrument-free, agar based method that has been modified to afford the simultaneous evaluation of as many as twelve bacterial strains. Due to its simplicity it can be applied to large collections of bacteria to rapidly screen for multi-drug resistant isolates that show an over-expression of their efflux systems. The principle of the method is simple and relies on the ability of the bacteria to expel a fluorescent molecule that is substrate for most efflux pumps, ethidium bromide. In this approach, the higher the concentration of ethidium bromide required to produce fluorescence of the bacterial mass, the greater the efflux capacity of the bacterial cells. We have tested and applied this method to a large number of Gram-positive and Gram-negative bacteria to detect efflux activity among these multi-drug resistant isolates. The presumptive efflux activity detected by the Ethidium Bromide-agar Cartwheel method was subsequently confirmed by the determination of the minimum inhibitory concentration for several antibiotics in the presence and absence of known efflux pump inhibitors.

scholarly journals Combining Normal/Reversed-Phase HPTLC with Univariate Calibration for the Piperine Quantification with Traditional and Ultrasound-Assisted Extracts of Various Food Spices of Piper nigrum L. under Green Analytical Chemistry Viewpoint

Molecules ◽  
2021 ◽  
Vol 26 (3) ◽  
pp. 732
Author(s):  
Mohammed H. Alqarni ◽  
Prawez Alam ◽  
Ahmed I. Foudah ◽  
Magdy M. Muharram ◽  
Faiyaz Shakeel
Keyword(s):  
Analytical Chemistry ◽  
High Performance ◽  
Black Pepper ◽  
Reversed Phase ◽  
Normal Phase ◽  
Piper Nigrum ◽  
Green Analytical Chemistry ◽  
Pharmaceutical Samples ◽  
Analytical Technique ◽  
Ultrasound Assisted

Due to unavailability of sustainable analytical techniques for the quantitation of piperine (PPN) in food and pharmaceutical samples, there was a need to develop a rapid and sensitive sustainable analytical technique for the quantitation of PPN. Therefore, the current research presents a fast and highly sensitive normal/reversed-phase high-performance thin-layer chromatography (HPTLC) technique with classical univariate calibration for the quantitation of PPN in various food spices of black pepper with traditional (TE) and ultrasound-assisted extracts (UBE) of various food spices of Piper nigrum L. under green analytical chemistry viewpoint. The amount of PPN in TE of four different spices of black pepper—namely BPMH, BPLU, BPSH, and BPPA—was found to be 309.53, 304.97, 282.82, and 232.73 mg g−1, respectively using a sustainable normal-phase HPTLC technique. However, the amount of PPN in UBE of BPMH, BPLU, BPSH, and BPPA was recorded as 318.52, 314.60, 292.41, and 241.82 mg g−1, respectively using a sustainable normal phase HPTLC technique. The greenness of normal/reversed-phase HPTLC technique was predicted using AGREE metric approach. The eco-scale was found to be 0.90, suggested excellent greenness of normal/reversed-phase technique. UBE of PPN was also found to be superior over TE of PPN. Overall, the results of this research suggested that the proposed normal/reversed-phase densitometry technique could be effectively used for the quantitation of PPN in food and pharmaceutical samples.

scholarly journals Secondary Metabolic Profile as a Tool for Distinction and Characterization of Cultivars of Black Pepper (Piper nigrum L.) Cultivated in Pará State, Brazil

2021 ◽  
Vol 22 (2) ◽  
pp. 890
Author(s):  
Luccas M. Barata ◽  
Eloísa H. Andrade ◽  
Alessandra R. Ramos ◽  
Oriel F. de Lemos ◽  
William N. Setzer ◽  
...  
Keyword(s):  
Black Pepper ◽  
Plant Diseases ◽  
Gas Chromatography Mass Spectrometry ◽  
Piper Nigrum ◽  
Volatile Components ◽  
Chemical Compositions ◽  
Multivariate Statistical ◽  
Protein Targets ◽  
Fruit Samples ◽  
Sesquiterpene Hydrocarbons

This study evaluated the chemical compositions of the leaves and fruits of eight black pepper cultivars cultivated in Pará State (Amazon, Brazil). Hydrodistillation and gas chromatography–mass spectrometry were employed to extract and analyze the volatile compounds, respectively. Sesquiterpene hydrocarbons were predominant (58.5–90.9%) in the cultivars “Cingapura”, “Equador”, “Guajarina”, “Iaçará”, and “Kottanadan”, and “Bragantina”, “Clonada”, and “Uthirankota” displayed oxygenated sesquiterpenoids (50.6–75.0%). The multivariate statistical analysis applied using volatile composition grouped the samples into four groups: γ-Elemene, curzerene, and δ-elemene (“Equador”/“Guajarina”, I); δ-elemene (“Iaçará”/“Kottanadan”/“Cingapura”, II); elemol (“Clonada”/“Uthirankota”, III) and α-muurolol, bicyclogermacrene, and cubebol (“Bragantina”, IV). The major compounds in all fruit samples were monoterpene hydrocarbons such as α-pinene, β-pinene, and limonene. Among the cultivar leaves, phenolics content (44.75–140.53 mg GAE·g−1 FW), the enzymatic activity of phenylalanine-ammonia lyase (20.19–57.22 µU·mL−1), and carotenoids (0.21–2.31 µg·mL−1) displayed significant variations. Due to black pepper’s susceptibility to Fusarium infection, a molecular docking analysis was carried out on Fusarium protein targets using each cultivar’s volatile components. F. oxysporum endoglucanase was identified as the preferential protein target of the compounds. These results can be used to identify chemical markers related to the susceptibility degree of black pepper cultivars to plant diseases prevalent in Pará State.

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