• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.403-411
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• 2005

The relationship between plasmid (~9.5 kb) and pediocin PA-1 in P. acidilactici K10 was confirmed by plasmid curing. The pediocin operon of P. acidilactici K10 was amplified by PCR (polymerase chain reaction), and the nucleotide sequence was analyzed. The sequence of the pediocin operon of P. acidilactici K10 was similar to those of P. acidilactici strains producing pediocin PA-1/ AcH. For the expression of pediocin PA-1 in E. coli, a pQEPED (pQE-30 Xa::mature pedA) was constructed. His-tagged recombinant pediocin PA-1 (-6.5 kDa) was translated by cell-free in vitro transcription and translation using pQEPED as a DNA template. Theresult of slot blotting assay showed that transcription of recombinant pedA in E. coli M15 was induced by the addition of isopropyl-$\beta$-D-thiogalactopyranoside (IPTG) at the final concentration of 1 mM. Although the recombinant pediocin PA-1 inhibited the growth of E. coli, it was expressed in the host strain and purified by nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography under denaturing condition. This is the first report for the production and one-step purification of biologically active recombinant pediocin PA-1 in E. coli.

• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.831-837
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• 2001

Pediocin K1, a bacteriocin produced by Pediococcus sp. K1 isolated from Korean traditional fermented flatfish, inhibited certain strains of Lactobacillus, Streptococcus, and Listeria monocytogenes. Pediocin K1 was found to be stable at $90^{\circ}C$ for 30 min. Among the organisms tested, Listeria monocytogenes was the most sensitive to pediocin K1 and was completely killed when the initial inoculum size of L.monocytogenes cells was equal to or less than $10^3 CFU/ml$. The degree of inibition of Listeria monocytogenes by pediocin K1 increased 10-fold on the addition of citric acid ($0.2\%$) to the medium, thereby showing the synergistic effect of citric acid. Listeria monocytogenes cells resistant to pediocin K1 appeared at a frequency of about $10^-4$/cells. Once developed after exposure to pediocin K1, the resistant phenotype still persisted in the absence of pediocin K1 in successive cultures. This infers that resistance may be attributable to genetic change(s) in the resistant cells.

• Food Science and Preservation
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• v.14 no.3
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• pp.328-331
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• 2007

This study was conducted to investigate the effect of pediocin treatment on noodle quality during 4 days of storage at $20^{\circ}C$. The pH of noodle increased after 2 days of storage and then decreased during further storage. The total bacterial counts in noodles increased during the storage period. When pediocin was present at 1,000 ppm, bacterial counts temporarily decreased after first day of storage and then slowly increased to 4 days of storage. Coliforms were detected after 2 days of storage in noodles stored without pediocin. When pediocin was present at 300 or 500 ppm, the coliform detection time was extended to 3 days of storage. Upon treatment with 1,000 ppm of pediocin, the coliform detection time was further extended to 4 days of storage. The fungal count in noodles was 2.3 log CFU/mL initially, and did not change significantly during the first day of storage, after which time the fungal count increased quickly. The fungal counts in noodles without pediocin treatment increased more rapidly than in noodles stored with pediocin, and was 5.0 log CFU/mL after 4 days of storage. We conclude that pediocin prevented noodle deterioration on storage.

• Food Science and Preservation
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• v.14 no.2
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• pp.131-135
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• 2007

We investigated the effects of pediocin on physicochemical and microbial changes in soybean cut 9 days of storage at $10^{\circ}C$, in order to improve shelf-life. As the storage time of curd increased the pH of solutions treated by pediocin immersion did not vary greatly, whereas the pH of control curd decreased after 5 days of storage. Titratable acidity increased in non-treated curd after 5 days of storage, and also in cud immersed in pediocin solutions of 300 and 500 ppm, after 6 and 7 days of storage, respectively. A pediocin solution of 1,000 ppm the development of titratable acidity. Also, turbidity did not increase during storage of curd treated with a pediocin solution of 1,000 ppm. The bacterial count of the immersion solution was $10^{2.5}$ CFU/mL at the commencement of storage, remained stable for 5 days of storage, and then increased rapidly. Coliforms were detected in untreated curd after 2.5days of storage. In curd treated with 300, 500 or 1,000ppm of pediocin, the elapsed times to coliform detection were 3.5 days, 5 days and 7 days, respectively, It is thus possible to prevent the deterioration of soybean curd with pediocin treatment.

• Journal of Microbiology and Biotechnology
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• v.13 no.3
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• pp.429-436
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• 2003

Pediocin K1 is a bacteriocin produced by Pediococcus sp. KCA 1303-10, isolated from traditionally fermented flatfish in Korea. Pediocin K1-dependent antibiosis and pediocin K1-independent antibiosis against Listeria monocyrogenes were investigated by comparing antibiosis potential of the ped＋ wild-type strain of Pediococcus sp. KCA1303-10 with that of the ped- mutant strain in 3 different media at 3 different temperatures. In the synthetic MRS-APT medium, bacteriocin (pediocin K1)-dependent antibiosis (BDA) acted as the major driving force of overall antibiosis at the initial stage before the pH of the media was not sufficiently lowered, while bacteriocin-independent antibiosis (BIA) took over the major role at the late stage of antibiosis by killing otherwise resistant cells in the modium. The role of BDA increased as the temperature of the system decreased. The antibiosis potential of BDA among the overall antibiosis of Pediococcus against Listeria at $37^{\circ}C$ was calculated as 46%, and as 75% at $25^{\circ}C$. In the skim milk medium, antibiosis of Pediococcus against Listeria was weakened more than 4 log cycles compared to that of the synthetic medium; however, BDA worked as the main antibiosis force regardless of the culturing temperature in the skim milk medium. In the bean soup medium, BDA also worked as the major killing mechanism against Listeria, but BIA played as another suppressing mechanism against otherwise pediocin-resistant Listeria population. These results suggest that a large portion of the inhibitory action of the ped+Pediococcus sp. KCA1303-10 was attributable to the bacteriocin produced by the strain and that viable Pediococcus sp. KCA1303-10 was superior to the purified bacteriocin in suppressing the occurrence of the bacteriocin-resistant Listeria monocytogenes in food systems.

• Journal of Microbiology and Biotechnology
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• v.20 no.8
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• pp.1215-1218
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• 2010

The recombinant DNA pLR5cat_PSAB, in which pediocin PA-1 structural and immunity genes (pedAB) fused with the promoter and deduced signal sequence of an ${\alpha}$-amylase gene from a bifidobacterial strain were inserted in Escherichia coli-lactobacilli shuttle vector pLR5cat, was transferred to Lactobacillus reuteri KCTC 3679 and the transformant presented bacteriocin activity. The recombinant L. reuteri KCTC 3679 transformed with the shortened pLR5cat(S)_PSAB, where a nonessential region for the lactobacilli replicon was removed, also showed bacteriocin activity. The molecular mass of the secreted pediocin PA-1 from the recombinant bacteria was the same as that of native pediocin PA-1 (~4.6 kDa) from Pediococcus acidilactici K10 on a sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel. In cocultures with Listeria monocytogenes, the recombinant L. reuteri KCTC 3679 effectively reduced the viable cell count of the pathogenic bacterium by a 3 log scale compared with a control where L. monocytogenes was incubated alone.

• Food Science of Animal Resources
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• v.30 no.1
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• pp.66-72
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• 2010

Pediococcus pentosaceus SA131 was isolated from jeotgal, is the bacteriocin producer against bovine mastitis pathogens, Streptococcus uberis E290, Enterococcus gallinarum E362, and Staphylococcus epidermidis ATCC 12228. The medium composition for pediocin SA131 production by P. pentosaceus SA131 was optimized using response surface methodology. Component of medium was studied as carbon source (glucose, fructose, lactose, glycerol, sucrose, maltose, and mannitol), nitrogen source (beef extract, yeast extract, peptone, malt extract, and tryptone), mineral and surfactant ($MgSO_4$, $KH_2PO_4$, $(NH_4)_2SO_4$, $MnSO_4$, NaCl, sodium acetate, and Tween 80). Through one factor-at-a-time experiment, glucose, fructose, yeast extract, malt extract, NaCl, $MgSO_4$, and Tween 80 were determined as the good ingredient. The effects of major factors for pediocin SA131 production were investigated by two-level fractional factorial designs (FFD). By a $2^4$ FFD, fructose, yeast extract, and $MnSO_4$ were found to be the important factors for the bacteriocin production. Subsequently, a $2^3$ central composite design (CCD) was adopted to derive a statistical model for optimizing the composition of the fermentation medium. The estimated optimum composition for the production of pediocin SA131 by P. pentosaceus SA131 was as follows; 0.13% fructose, 1% glucose, 1.8% yeast extract, 2.58% $MnSO_4$, 0.2% NaCl, and 0.2% Tween 80. The pediocin production under optimized medium was increased to 1,000 AU/mL, compared to the 400 AU/mL in MRS medium.

• Preventive Nutrition and Food Science
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• v.15 no.4
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• pp.360-363
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• 2010

To enhance the expression and secretion of pediocin PA-1 from heterologous bacterial hosts, the promoter and deduced signal sequence (PS) of an $\alpha$-amylase gene from a Bifidobacterium adolescentis strain was fused with pediocin PA-1 structural and immunity genes (AB) and the resulting functions were evaluated in Escherichia coli. Two recombinant PCR products were created-one with just the deduced signal sequence and one with the sequence plus the Ser and Thr sequences that are the next two amino acids of the signal sequence. These two products, the PSAB (---AQA::KYY---) and PSABST (---AQA$\underline{ST}$::KYY---), respectively, were inserted into a TA cloning vector (yT&A) and named pPSAB, which was previously reported, and pPSABST. The two recombinant plasmid DNAs were transferred into E. coli JM109 and the transformants displayed antimicrobial activity, where the activity of E. coli JM109 (pPSAB) was stronger than that of E. coli JM109 (pPSABST), indicating that the ST amino acid residues were not necessary for secretion and might have even decreased the antimicrobial activity of recombinant pediocin PA-1.

• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.96-105
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• 2002

A bacteriocin-producing strain identified as Pediococcus acidilactici was isolated from kimchi. The bacteriocin was identified to belong to the pediocin family and exhibited bactericidal activity against most Gram-positive bacteria as well as some Gram-negative bacteria. The bacteriocin was stable up to $80^{\circ}C with wide pH ranges (5.0-10.0). The bactericidal activity remained unchanged after treatment with nonproteolytic enzymes such as nuclease and ${\alpha}$-amylase, however, it was destroyed after treatment with protease. The bacteriocin was effectively extracted by the pH-mediated adsorption-desorption method and purified effectively by semi-preparative RP-HPLC. The molecular weight of the bacteriocin was 4,622, as determined by electrospray mass spectrometry. The amino acid sequence consisted of 44 amino acid residues with four cysteines. The high solubility and pH stability of the isolated pediocin provide definite advantages over nisin and other bacteriocins in regards to its potential applications.

• Proceedings of the Microbiological Society of Korea Conference
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• 2005.05a
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• pp.215.4-215
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• 2005