Antibacterial activity produced by Enterococcus spp. isolated from an artisanal Mexican dairy product, Cotija cheese

García‐Cano, Israel; Serrano-Maldonado, Carlos Eduardo; Olvera-García, Myrna; Delgado-Arciniega, Estela; Peña-Montes, Carolina; Mendoza‐Hernández, Guillermo; Quirasco, Maricarmen

doi:10.1016/j.lwt.2014.04.059

Cited by 28 publications

(21 citation statements)

References 35 publications

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“…The recombinant protein was active only against gram-positive bacteria, in contrast with the previous report by García-Cano et al [2014], where the unpurified enzyme preparation showed activity against gram-nega-DOI: 10.1159/000486757 tive bacteria as well. This result suggests that one or more of the rest of the enzymes with PGH activity visualized by zymography in that report could be responsible for the activity against gram-negative bacteria.…”

Section: Discussioncontrasting

confidence: 95%

“…The recombinant AtlD produced in E. coli is up to 36 times more active versus S. aureus and Listeria species than the unpurified enzyme preparation from the native producing strain [García-Cano et al, 2014], which constitutes an interesting issue for a potential biotechnological application of the recombinant enzyme in food safety. Finally, it displayed lytic activity against the pathogenic strain E. faecalis V583, and other enterococcal strains of clinical origin, which are characterized as being highly resistant to antibiotics.…”

Section: Discussionmentioning

confidence: 99%

“…Of these, only the following 3 had been previously characterized: AtlA, which shows N-acetylglucosaminidase activity with a molecular weight of 72 kDa and another active form of 62 kDa [Eckert et al, 2006], and AtlB and AtlC, both with an N-acetylmuramidase, lysozyme-like activity and molecular weights of 50 and 47 kDa, respectively [Mesnage et al, 2008]. In our research group, we reported the identification of a PGH with 2 lytic domains (N-acetylglucosaminidase and CHAP) produced by E. faecalis F, a strain isolated from Cotija cheese -a Mexican handmade ripened cheese -that had not been reported before [García-Cano et al, 2014]. In order to contribute to the knowledge of E. faecalis PGH, this new enzyme, named AtlD from now on, was cloned and expressed in E. coli.…”

Section: Introductionmentioning

confidence: 86%

“…Bacterial Strains and Growth Conditions E. faecalis F was isolated form a Mexican ripened, artisanally made cheese, i.e., Cotija cheese [García-Cano et al, 2014], and it was cultivated in MRS (de Man, Rogosa, Sharpe) broth (Oxoid, Basingstoke, UK) at 37 ° C and 250 rpm until the early stationary phase. Escherichia coli DH5α and E. coli BL21 (DE3) (Invitrogen, Carlsbad, CA, USA) were used as cloning and expression strains, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…It was identified by LC-MS/MS as a putative N-acetylglucosaminidase [García-Cano et al, 2014]. In order to know the sequence of the PGH coding gene, a longer DNA region of 2,076 bp, containing the PGH open reading frame (ORF) as well as the flanking ORF, was amplified and sequenced ( Fig.…”

Section: Pgh Gene Sequencementioning

confidence: 99%

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Cloning and Characterization of a Novel N-acetylglucosaminidase (AtlD) from Enterococcus faecalis

et al. 2018

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The atlD gene from an Enterococcus faecalis strain isolated from a Mexican artisanal cheese was cloned, sequenced and expressed in Escherichia coli in order to perform a biochemical characterization. A partial amino acid sequence of the heterologous protein was obtained by LC-MS/MS, and it corresponded to a novel peptidoglycan hydrolase designated AtlD. Its molecular mass was 62–75 kDa, as determined by SDS-PAGE, zymography, Western blot, and exclusion chromatography. Electrofocusing rendered an isoelectric point (pI) of 4.8. It exhibited N-acetylglucosaminidase activity, with an optimal pH and temperature between 6–7 and 50°C, respectively. It retained 85% activity with NaCl at 1,000 mM, but it was susceptible to divalent ions, particularly Zn²⁺. It showed antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, and enterococcal strains of clinical origin. Due to the fact that it showed activity versus pathogenic bacteria, and because of its capabilities under ionic strength, temperature, and pH values present in food matrices, it could be applied as an additive in the food industry. This study will aid in the design of new antibacterial agents of natural origin to combat food-borne diseases, and it could be used as an industrial or hospital hygiene agent as well.

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Section: Discussioncontrasting

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 86%

Section: Methodsmentioning

confidence: 99%

Section: Pgh Gene Sequencementioning

confidence: 99%

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Cloning and Characterization of a Novel N-acetylglucosaminidase (AtlD) from Enterococcus faecalis

et al. 2018

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Electrochemical/Voltammetric/Amperometric Nanosensors for the Detection of Pathogenic Bacteria

Ahmed

Patel

2023

Nanosensors for Point-of-Care Diagnostics of Pathogenic Bacteria

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Expression, purification, and characterization of a bifunctional 99-kDa peptidoglycan hydrolase from Pediococcus acidilactici ATCC 8042

García‐Cano

Campos-Gómez

Contreras-Cruz

et al. 2015

Appl Microbiol Biotechnol

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Pediococcus acidilactici ATCC 8042 is a lactic acid bacteria that inhibits pathogenic microorganisms such as Staphylococcus aureus through the production of two proteins with lytic activity, one of 110 kDa and the other of 99 kDa. The 99-kDa one has high homology to a putative peptidoglycan hydrolase (PGH) enzyme reported in the genome of P. acidilactici 7_4, where two different lytic domains have been identified but not characterized. The aim of this work was the biochemical characterization of the recombinant enzyme of 99 kDa. The enzyme was cloned and expressed successfully and retains its activity against Micrococcus lysodeikticus. It has a higher N-acetylglucosaminidase activity, but the N-acetylmuramoyl-L-alanine amidase can also be detected spectrophotometrically. The protein was then purified using gel filtration chromatography. Antibacterial activity showed an optimal pH of 6.0 and was stable between 5.0 and 7.0. The optimal temperature for activity was 60 °C, and all activity was lost after 1 h of incubation at 70 °C. The number of strains susceptible to the recombinant 99-kDa enzyme was lower than that susceptible to the mixture of the 110- and 99-kDa PGHs of P. acidilactici, a result that suggests synergy between these two enzymes. This is the first PGH from LAB that has been shown to possess two lytic sites. The results of this study will aid in the design of new antibacterial agents from natural origin that can combat foodborne disease and improve hygienic practices in the industrial sector.

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Antibacterial activity produced by Enterococcus spp. isolated from an artisanal Mexican dairy product, Cotija cheese

Cited by 28 publications

References 35 publications

Cloning and Characterization of a Novel N-acetylglucosaminidase (AtlD) from Enterococcus faecalis

Cloning and Characterization of a Novel N-acetylglucosaminidase (AtlD) from Enterococcus faecalis

Electrochemical/Voltammetric/Amperometric Nanosensors for the Detection of Pathogenic Bacteria

Expression, purification, and characterization of a bifunctional 99-kDa peptidoglycan hydrolase from Pediococcus acidilactici ATCC 8042

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