Extraction, partial purification and characterization of a bacteriocin (fragilicin) produced by a strain of Bacteroides fragilis isolated from Callithrix penicillata

Farias, Luiz M.; Tótola, Antônio Helvécio; Miranda, Cláudia Madeira; Carvalho, Maria Auxiliadora Roque de; Damasceno, Carlos Américo Veiga; Tavares, Carlos Alberto Pereira; Cisalpino, E. O.; Vieira, Eliane

doi:10.1016/0923-2508(94)90062-0

Cited by 22 publications

(22 citation statements)

References 26 publications

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“…Cells were broken by the Mini BeadBeater-8 (BioSpec Products, Inc., Bartlesville, OK) in PBS buffer, and broken cells were centrifuged at 6,000g for 10 min. Supernatants were evaluated for antimicrobial activity using L. monocytogenes Scott A as indicator strain, using the double-layer diffusion assay [16]. The molecular mass of the cloned peptides was confirmed by mass spectrometry, as described previously.…”

Section: Mass Spectrometrymentioning

confidence: 99%

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Antimicrobial compounds produced by Lactobacillus sakei subsp. sakei 2a, a bacteriocinogenic strain isolated from a Brazilian meat product

Carvalho

Bambirra

Kruger

et al. 2009

J Ind Microbiol Biotechnol

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Bacteriocins produced by lactic acid bacteria are gaining increased importance due to their activity against undesirable microorganisms in foods. In this study, a concentrated acid extract of a culture of Lactobacillus sakei subsp. sakei 2a, a bacteriocinogenic strain isolated from a Brazilian pork product, was purified by cation exchange and reversed-phase chromatographic methods. The amino acid sequences of the active antimicrobial compounds determined by Edman degradation were compared to known protein sequences using the BLAST-P software. Three different antimicrobial compounds were obtained, P1, P2 and P3, and mass spectrometry indicated molecular masses of 4.4, 6.8 and 9.5 kDa, respectively. P1 corresponds to classical sakacin P, P2 is identical to the 30S ribosomal protein S21 of L. sakei subsp. sakei 23 K, and P3 is identical to a histone-like DNA-binding protein HV produced by L. sakei subsp. sakei 23 K. Total genomic DNA was extracted and used as target DNA for PCR amplification of the genes sak, lis and his involved in the synthesis of P1, P2 and P3. The fragments were cloned in pET28b expression vector and the resulting plasmids transformed in E. coli KRX competent cells. The transformants were active against Listeria monocytogenes, indicating that the activity of the classical sakacin P produced by L. sakei 2a can be complemented by other antimicrobial proteins.

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Section: Mass Spectrometrymentioning

confidence: 99%

“…The following strains were used (Table 1) Assay for antibacterial activity Antibacterial activity was assayed by the double layer diffusion test, according to Farias et al [16]. Ten microliters of the testing material was spotted onto the surface of plates containing BHI agar (Difco).…”

Section: Bacterial Cultures and Mediamentioning

confidence: 99%

Antimicrobial compounds produced by Lactobacillus sakei subsp. sakei 2a, a bacteriocinogenic strain isolated from a Brazilian meat product

Carvalho

Bambirra

Kruger

et al. 2009

J Ind Microbiol Biotechnol

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“…mutans in prevention of dental caries (Hillman 2002). In contrast, there are very few studies regarding bacteriocins that are produced by oral Gram-negative bacteria (Takada et al 1991;Höhne et al 1993;Farias et al 1994;Oliveira et al 1998). Prevotella intermedia (Takada et al 1991), Bacteroides melaninogenicus (Nakamura et al 1981), and Actinobacillus actinomycetemcomitans have been reported to produce bacteriocins; however, their genetic and biochemical properties have not been characterized.…”

Section: Introductionmentioning

confidence: 99%

Characterization of a bacteriocin produced by Prevotella nigrescens ATCC 25261

Kaewsrichan

Douglas

Nissen‐Meyer

et al. 2004

Lett Appl Microbiol

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Aims: To characterize the antimicrobial activity produced by Prevotella nigrescens ATCC 25261, and to evaluate its safety on cultured gingival fibroblasts. Methods and Results: An antimicrobial activity was obtained from purifying the culture supernatant of Pr. nigrescens ATCC 25261. Purification of the active compound was achieved with ammonium sulphate precipitation followed by anion-exchange and gel filtration chromatography. As revealed by SDS-PAGE, the active fraction was relatively homogeneous, showing a protein with an approximate molecular weight of 41 kDa. The antimicrobial compound, named nigrescin, exhibited a bactericidal mode of action against Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, and Actinomyces spp. Nigrescin was stable in a pH range between 6AE5 and 9AE5, at 100°C for 10 min, and resistant to lyophilization. But its activity was lost after proteinase K treatment. Despite at very high concentrations beyond the minimum inhibitory concentration (MIC), nigrescin was not toxic to the gingival fibroblasts. Conclusion: Nigrescin is a novel bacteriocin produced by Pr. nigrescens ATCC 25261. It exhibits antimicrobial activity against species that are implicated in periodontal diseases. The absence of toxicity on the gingival fibroblasts suggests the possibility in using of nigrescin for an application in periodontal treatment. Significance and Impact of the Study: Novel evidence on nigrescin would make Pr. nigrescens ATCC 25261 attractive in biotechnological applications as an antimicrobial agent in clinical dentistry.

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“…However, several authors have reported about bacteriocins having a broad spectrum of action [15,30,10,2]. This is also the case for the bacteriocin-like substances produced by A. actinomycetemcomitans P 7.20 [22] that was submitted to partial puriWcation and characterization processes.…”

Section: Discussionmentioning

confidence: 93%

Actinomycetemcomitin: a new bacteriocin produced by Aggregatibacter (Actinobacillus) actinomycetemcomitans

Lima¹,

Carvalho²,

Apolônio³

et al. 2007

J Ind Microbiol Biotechnol

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Aggregatibacter (Actinobacillus) actinomycetemcomitans P(7-20) strain isolated from a periodontally diseased patient has produced a bacteriocin (named as actinomycetemcomitin) that is active against Peptostreptococcus anaerobius ATCC 27337. Actinomycetemcomitin was produced during exponential and stationary growth phases, and its amount decreased until it disappeared during the decline growth phase. It was purified by ammonium sulphate precipitation (30-60% saturation), and further by FPLC (mono-Q ionic exchange and Phenyl Superose hydrophobic interaction) and HPLC (C-18 reversed-phase). This bacteriocin loses its activity after incubation at a pH below 7.0 or above 8.0, following heating for 30 min at 45 degrees C, and after treatment with proteolytic enzymes such as trypsin, alpha-chymotrypsin, and papain. Actinomycetemcomitin has a molecular mass of 20.3 KDa and it represents a new bacteriocin from A. actinomycetemcomitans.

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Extraction, partial purification and characterization of a bacteriocin (fragilicin) produced by a strain of Bacteroides fragilis isolated from Callithrix penicillata

Cited by 22 publications

References 26 publications

Antimicrobial compounds produced by Lactobacillus sakei subsp. sakei 2a, a bacteriocinogenic strain isolated from a Brazilian meat product

Antimicrobial compounds produced by Lactobacillus sakei subsp. sakei 2a, a bacteriocinogenic strain isolated from a Brazilian meat product

Characterization of a bacteriocin produced by Prevotella nigrescens ATCC 25261

Actinomycetemcomitin: a new bacteriocin produced by Aggregatibacter (Actinobacillus) actinomycetemcomitans

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