Detection of specific bacteriocin-producing lactic acid bacteria by colony hybridization

Martínez, Vicente; Rodríguez,; Medina,; Hernández,

doi:10.1046/j.1365-2672.1998.00448.x

Cited by 20 publications

(6 citation statements)

References 13 publications

(17 reference statements)

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“…The methods described by Grunstein and Hogness [20] consists in extracting the DNA from bacterial colonies and to bind it to the Hybond-N nylon membrane in situ [28]. The resulting membrane was successively tested with different probes, obtained by PCR using the species-specific primers (Table 1) for reference strains Lactobacillus paracasei DSM 5622 T , Lact.…”

Section: Colony Hybridization Analysismentioning

confidence: 99%

Genotypic characterization of non starter lactic acid bacteria involved in the ripening of artisanal Bitto PDO cheese

Colombo

Borgo

Fortina

2009

J. Basic Microbiol.

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Bitto of Valchiavenna, an artisanal Italian cheese produced without the addition of any starter cultures, has been attributed a protected designation origin (PDO) cheese, but the strain composition of the natural microbial population colonizing this traditional dairy product is still unknown. To obtain preliminary information on the non starter lactic acid bacteria involved in its ripening, a total of 136 NSLAB isolates, randomly selected from MRS and M17 agar plates, were collected from three different cheese samples after 120 days of ripening. The new isolates were identified by combining PCR 16S-23S rDNA spacer analyses, partial 16S rRNA gene sequencing, species-specific probes and colony hybridization. Eighty-two isolates, representing 60% of the total strains selected, were homofermentative cocci: 83% of them were enterococci, with Enterococcus durans being the predominant species found. Pediococcus spp. were also isolated, together with strains of Streptococcus thermophilus. Within lactobacilli, 57% of the isolates were identified as Lactobacillus paracasei; Lact. curvatus, Lact. plantarum, Lact. fermentum, were present in a lower amount. The isolates were differentiated at strain-level by Rep-PCR analysis. This is the first effort to microbiological characterization of Valchiavenna's Bitto; the results suggest the possibility of preserving the wild bacterial population in order to protect the typical organoleptic characteristics of this traditional raw milk cheese and to select new strains for the dairy industry.

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Section: Colony Hybridization Analysismentioning

confidence: 99%

Genotypic characterization of non starter lactic acid bacteria involved in the ripening of artisanal Bitto PDO cheese

Colombo

Borgo

Fortina

2009

J. Basic Microbiol.

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“…Genetic methods such as polymerase chain reaction (PCR) (Klijn et al. 1995) and colony hybridization (Martínez et al. 1998) were also attempted.…”

Section: Introductionmentioning

confidence: 99%

Bacteriocin detection by liquid chromatography/mass spectrometry for rapid identification

et al. 2007

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Aims: To establish a new system to detect and identify bacteriocins in the early stage of screening for novel bacteriocins. Methods and Results: Liquid chromatography/mass spectrometry (LC/MS) was employed for development of a new system for rapid detection and identification of bacteriocins. The system detected and identified bacteriocins such as nisin and lacticin 481 from 25 μl of culture supernatants of their producing strains by accurate mass determination coupled with simultaneous impurity removal within 40 min. Especially, the system clearly distinguished three nisin variants (A, Z, Q) in culture supernatants of their producing strains, although they have similar structures and molecular masses. Each one‐step pretreatment by cell adsorption–desorption or acetone precipitation improved bacteriocin detection dramatically, especially for mundticin KS. This system could be applied for detection and molecular mass determination of novel bacteriocins by extracting bacteriocin‐related ions. Conclusions: The developed system could detect and identify some kinds of bacteriocin from culture supernatants or pretreated samples. Significance and Impact of the Study: The developed system helps us to identify bacteriocins in the early stage of screening without any or with one‐step pretreatment. This system is effective on not only detection of known bacteriocins but also identification of novel bacteriocins. Consequently, this system will accelerate discovery of novel bacteriocins.

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“…Previously, Bhunia et al (1994) had demonstrated the use of the Southern hybridization technique for the identification of bacteriocin-encoding plasmids. DNA probes have been used earlier to detect silage colonization by Lactobacillus and Pediococcus strains and bacteriocinogenic LAB (Rodriguez et al 1997;Martinez et al 1998). In these studies, the specificity of the DNA probe was tested using pure cultures, but in the present investigation, the pediocin PA-1 probe has been used directly to screen a heterogeneous population of antagonistic cultures isolated from naturally fermented cucumber.…”

Section: Rapid Detection Of Pediocin Producers By Colony Hybridizationmentioning

confidence: 98%

“…Rapid methods such as colony immuno-blot, PCR, gene probe, etc. have been employed that help to overcome these inconveniences (Rodriguez et al 1997;Martinez et al 1998). In the recent past, production of intergeneric and interspecific pediocin PA-1/AcH by Lactobacillus plantarum and P. parvulus has been reported (Ennahar et al 1996;Bennik et al 1997).…”

Section: Introductionmentioning

confidence: 98%

Fermenting Cucumber, a Potential Source for the Isolation of Pediocin-Like Bacteriocin Producers

Halami

Ramesh

Chandrashekar

2005

World J Microbiol Biotechnol

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A strain of Pediococcus acidilactici CFR K7 isolated from cucumber, produced an antimicrobial peptide which acted against Leuconostoc mesenteroides, selected strains of Lactobacillus spp., Pediococcus spp. and Enterococcus spp. The partially purified bacteriocin had molecular weight of $4.6 kDa, heat stability in a range of 40-121°C and was active over a wide range of pH (2.0-9.0). This bacteriocin possessed strong antilisterial activity and was susceptible to proteolytic enzymes. Southern hybridization using the PCR-generated pedA probe established that the gene for the bacteriocin was plasmid-borne as in the case of pediocin PA-1. Nucleotide sequence of the pedAB gene indicated 100% homology to a pediocin AcH/PA-1. Certain bacteriocinogenic strains isolated from naturally fermented cucumber were tested by colony hybridization using the pedA gene probe. Nine out of twenty colonies reacted with the probe indicating their ability to produce the pediocin-like bacteriocin. These nine colonies were further tested for their antimicrobial spectrum, proteolytic inactivation and plasmid profile. It was found that a few of them were active against Bacillus cereus, Micrococcus luteus and Listeria monocytogenes. Their proteolytic inactivation showed that the antimicrobial compound was susceptible to proteinase K. Colony hybridization could thus enable rapid detection of pediocin and pediocin-like bacteriocin producers among a population of bacteriocinogenic strains.

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Detection of specific bacteriocin-producing lactic acid bacteria by colony hybridization

Cited by 20 publications

References 13 publications

Genotypic characterization of non starter lactic acid bacteria involved in the ripening of artisanal Bitto PDO cheese

Genotypic characterization of non starter lactic acid bacteria involved in the ripening of artisanal Bitto PDO cheese

Bacteriocin detection by liquid chromatography/mass spectrometry for rapid identification

Fermenting Cucumber, a Potential Source for the Isolation of Pediocin-Like Bacteriocin Producers

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