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Isolation and Characterization of Pediocin Producing Pediococcus pentosaceus Pep1 from Vacuum-Packed Sausages
Abstract
Pediococcus pentosaceus Pep1 was isolated from vacuum-packed sausages, and it was shown to produce a potentially novel antimicrobial agent active against several species of Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, Staphylococcus, Enterococcus, Bacillus and Listeria, many of which are associated with food spoilage and health hazards of food origin. Gram- negative organisms such as Yersinia enterocolitica and Escherichia coli were not inhibited by this antimicrobial agent. Since the antimicrobial agent was found to be sensitive to proteolytic enzymes, resistant to heat and organic solvents and active over a wide range of pH values, between 3 and 8, apparently most stable in the lower part of that range, it was identified as a bacteriocin and was designated as pediocin P.
... P. pentosaceus was isolated at two of the production sites and three processing stages or time points. P. pentosaceus is homofermentative has antimicrobial and antioxidant properties, and is often used as a starter culture bacterium for fermenting foods with good bio-preservation characteristics [84][85][86]. P. pentosaceus can tolerate low pH/acids and bile salts, improve safety and quality, extend shelf life, has anti-mycotoxin effect, and affect the flavour characteristics of food products [84][85][86]. This bacterium has been associated with the fermentation of cereal-based foods such as borde from Ethiopia [61] and dèguè from Burkina Faso [87]. ...
... P. pentosaceus is homofermentative has antimicrobial and antioxidant properties, and is often used as a starter culture bacterium for fermenting foods with good bio-preservation characteristics [84][85][86]. P. pentosaceus can tolerate low pH/acids and bile salts, improve safety and quality, extend shelf life, has anti-mycotoxin effect, and affect the flavour characteristics of food products [84][85][86]. This bacterium has been associated with the fermentation of cereal-based foods such as borde from Ethiopia [61] and dèguè from Burkina Faso [87]. ...
Spontaneous fermentation of cereals like millet involves a diverse population of microbes from various sources, including raw materials, processing equipment, fermenting receptacles, and the environment. Here, we present data on the predominant microbial species and their succession at each stage of the Hausa koko production process from five regions of Ghana. The isolates were enumerated using selective media, purified, and phenotypically characterised. The LAB isolates were further characterised by 16S rRNA Sanger sequencing, typed using (GTG)5 repetitive-PCR, and whole genome sequencing, while 28S rRNA Sanger sequencing was performed for yeast identification. The pH of the millet grains ranged from mean values of 6.02—6.53 to 3.51—3.99 in the final product, depending on the processors. The mean LAB and yeast counts increased during fermentation then fell to final counts of log 2.77–3.95 CFU/g for LAB and log 2.10–2.98 CFU/g for yeast in Hausa koko samples. At the various processing stages, the counts of LAB and yeast revealed significant variations (p < 0.0001). The species of LAB identified in this study were Limosilactobacillus pontis, Pediococcus acidilactici, Limosilactobacillus fermentum, Limosilactobacillus reuteri, Pediococcus pentosaceus, Lacticaseibacillus paracasei, Lactiplantibacillus plantarum, Schleiferilactobacillus harbinensis, and Weissella confusa. The yeasts were Saccharomyces cf. cerevisiae/paradoxus, Saccharomyces cerevisiae, Pichia kudriavzevii, Clavispora lusitaniae and Candida tropicalis. The identification and sequencing of these novel isolates and how they change during the fermentation process will pave the way for future controlled fermentation, safer starter cultures, and identifying optimal stages for starter culture addition or nutritional interventions. These LAB and yeast species are linked to many indigenous African fermented foods, potentially acting as probiotics in some cases. This result serves as the basis for further studies into the technological and probiotic potential of these Hausa koko microorganisms.
... But able to grow at pH (4.5-8). (1) This bacteria grow more stably at more acidic PH (4).Pediococcus pentosaceus are unique in that they form tetrads (1). Pediococcus pentosaceus can produce antimicrobial agents known as bacteriocin (3) agants several species of Lactobacillus, Lactococcus, Leuconostic, Pediococcus, Staphylococcus aureus, Bacillus and Listeria (4). ...
... Pediococcus pentosaceus can produce antimicrobial agents known as bacteriocin (3) agants several species of Lactobacillus, Lactococcus, Leuconostic, Pediococcus, Staphylococcus aureus, Bacillus and Listeria (4). The bacteriocin isolated in Pediococcus pentosaceus was labeled pediocin (4). The end product of fermentation is lactic acid which lowers the environmental pH. ...
About 10 isolates of Pediococcus sp were isolated from different cheese made in Iraq, These isolates were identified morphologically and biochemically and Api20 kit, thus there was only 6 isolate were identified as Pediococcus pentosaceus (60%).In this study, we investigate, the effect of crude Bacteriocin from Pediococcus pentosaceus on 30 clinical isolates (5 E.coli, 5 Klepsiella pneumoniae, 5 Staphylococcus aureus, 5 Pseudomonas aeroginosa, 5 Bacillus subtilis, 5 Candida albicans). The protein concentration of this Bacteriocin was measured 67mg\ml by Bradford method and used as (1:2) by vol during the measuring the antimicrobial activity against the above clinical isolates by two methods wells and agar plug assay. The results showed that the inhibitory activity of this Bacteriocin was higher by wells method than agar pluq assay against Gram–positive bacteria or Gram-negative bacteria and yeast under this study.
... Recent studies have reported antilisterial activity of Pediococcus spp. from various sources (23,24). Pediococcus spp. ...
... A major antimicrobial substance from Pediococcus spp. has been found to be a bacteriocin called pediocin, which is classifi ed into Class II (24). Its molecular size is less than 5 kDa containing 36-48 residues (25). ...
The objective of this study is to evaluate the antilisterial effect of Pediococcus pentosaceus T1, which was isolated from kimchi, and to assess its potential for extending the shelf life of salmon and kimchi. Pediococcus pentosaceus T1 culture effectively inhibited proliferation of Listeria monocytogenes in a dose-dependent manner in a salmon-based medium. Antilisterial effect of the culture was stronger than that of nisin, an antibacterial peptide, as evidenced by lower minimum inhibitory concentration value (20 mg/mL) compared to nisin (over 20 mg/mL). P. pentosaceus T1 culture also effectively inhibited the growth of Listeria in salmon fillet. In particular, the culture (6 g per 100 mL) showed a stronger inhibitory effect than sodium hypochlorite (0.2 mg/mL), a disinfectant used in food processing. In kimchi fermentation, the treatment with P. pentosaceus T1 culture suppressed changes of acidity and pH during maturation. The inhibitory effect of the culture on kimchi lactic acid bacteria, which include Leuconostoc mesenteroides and Lactobacillus sakei, led to a drastic decrease in maturation rates of kimchi. Moreover, sensory test on kimchi treated with P. pentosaceus T1 showed that the culture improved overall acceptability of kimchi, which can be observed in higher scores of sourness, texture, off-flavour and mouthfeel compared with untreated kimchi. The results of this study suggest that kimchi-derived P. pentosaceus T1 could be a potential antilisterial agent in fish products as well as a starter to control overmaturation of kimchi.
... 다고 보고되어 있다(Fleming et al., 1975; Cintas et al., 1998; Osmanagaoglu et al., 2001; Wu et al., 2004). Pediocin은 저분 자의 내열성을 지니는 Class IIa bacteriocin으로 표적 세균의 세포벽 합성을 억제하거나 세포막에 구멍을 형성하여 살균 효 과를 나타내며(Sablon et al., 2000: Hechard and Sahl, 2002: Bauer and Dicks, 2005), pediocin을 암호화하는 유전자는 일 반적으로 plasmid DNA에 operon 형태로 존재한다고 보고되 어 있다(Miller et al., 2005). ...
... 백합(Meretrix meretrix) 식해에서 분리한 Pediococcus pentosaceus SH-10에 의한 병원성 세균의 억제 기작 , 1987), 위장관내 병원균 의 증식억제(Fernandes et al., 1992), 혈중 콜레스테롤의 저하 (Suzuki and Kaizu, 1991), 특이 및 비특이 면역반응의 유도 및 영양소 이용성의 향상(Fernandes et al., 1992), 암 퇴화 및 장 내효소 활성감소에 의한 결장암의 예방효과(Goldin and Gorbach, 1984생균제 미생물은 유산균이 주종을 이루고 있는 실정이다. 국내 의 생균제에 대한 최근 연구는 한국인의 장 내용물 또는 전통 발효식품으로부터 우수한 유산균을 분리하여 이를 식품이나 의약품으로 이용하고자 하는 노력이 활발하게 진행되고 있는 실정이다 (Kang et al., 2001; Ha et al., 2004; Kim et al., 2005; Shin et al., 2008; Lim and Im, 2009)et al., 1975; Cintas et al., 1998; Osmanagaoglu et al., 2001; Wu et al., 2004) ...
In this study, we investigated the mechanism of inhibition of pathogenic bacteria by Pediococcus pentosaceus strain SH-10 isolated from hard Clam Meretrix meretrix sikhae. When P. pentosaceus SH-10 was co-cultured in MRS broth with pathogenic bacteria, including Bacillus cereus, Listeria monocytogenes, Salmonella choleraesuis and Staphyloccus aureus, no viable pathogenic cells were detected after 18 h of incubation. However, pediocin or a pediocin-like bacteriocin was not detected in cultures of P. pentosaceus SH-10 by the agar diffusion method. Organic acids were produced in MRS broth in proportion to the incubation time of P. pentosaceus SH-10. These results indicate that P. pentosaceus SH-10 inhibited the growth of pathogenic bacteria by lowering the pH of the growth medium through the production of organic acids, including sodium lactate, sodium acetate, and sodium citrate.
... A higher temperature was selected as the optimum for pediocin production for P. acidilactici F (a sausage isolate) [69]. Production of pediocins from P. pentosaceus strains was at 37°C for P. pentosaceus ACCEL [70], and P. pentosaceus L and S (isolated from pork meat) [71] , 35°C for P. pentosaceus Pep1 (isolated from sau- sages) [72], 30°C for P. pentosaceus ST18 (isolated from boza drink) [73] and P. pentosaceus SM-1 (isolated from pork sausage) [56] . Pediocin PD-1 from Pediococcus damnosus NCFB1832 [54] is produced at 30°C. ...
... They are sensitive to most proteases. These characteristics are common in a number of pediocins as shown with pediocin AcH/PA-1 [5] , pediocin ST18 [73], pediocin AcM [104], pediocin L50 [103], pediocin PD-1 [22], pediocin F [69] , the pediocins produced by P. pentosceus Pep1 [72] and P. pentosaceus K23-2 [107].Table 2 summarizes some of the properties of isolated pediocins, in the cases the particular characteristics were examined and allow comparisons. The obvious differences , as shown inTable 2, may result in differences in the way of action and may explain the differences found in the inhibitory spectra of various pediocins. ...
Class IIa bacteriocins from lactic acid bacteria are small, cationic proteins with antilisterial activity. Within this class, the pediocins are those bacteriocins that share a highly conserved hydrophilic and charged N-terminal part harboring the consensus sequence -YGNGV- and a more variable hydrophobic and/or amphiphilic C-terminal part. Several pediocins have been isolated and characterized. Despite the structural similarities, their molecular weight varies, as well as their spectrum of antimicrobial activity. They exhibit important technological properties, e.g. thermostability and retaining of activity at a wide pH range, which along with the bactericidal action against Gram-positive food spoilage and pathogenic bacteria, make them an important class of biopreservatives. Much new information regarding the pediocins has emerged during the last years. In this review, we summarize and discuss all the available information regarding the sources of pediocins, the characteristics of their biosynthesis and production in fermentation systems, the characteristics of the known pediocin molecules, and their antibacterial action. The advances made by genetic engineering in improving the features of pediocins are also discussed, as well as their perspectives for future applications.
... The pediocins described to date include PA-1 (Gonzalez and Kunka 1987), AcH (Bhunia et al. 1988), JD (Richter et al. 1989), SJ-1 (Schved et al. 1993), 5 (Huang et al. 1996), AcM by Pediococcus acidilactici isolated from fermented meat (Elegado et al. 1997) and PD-1 by Pediococcus damnosus NCFB 1832 isolated from spoiled beer (Green et al. 1997). Bacteriocins produced by different strains of Pediococcus pentosaceus have also been reported, including pediocin A (Etchells et al. 1964;Fleming et al. 1975), pediocin N5p (Strasser de Saad and Manca de Nadra 1993), pediocin ACCEL (Wu et al. 2004 and pediocin P (Osmanagaoglu et al. 2001). Pediocins are considered potential food biopreservatives owing to their antimicrobial activity against some food-spoilage and pathogenic bacteria, such as Listeria monocytogenes and Staphylococcus aureus (Cintas et al. 1998). ...
... The bacteriocin was stable over a wide range of pH (2-8), especially in more acidic conditions. Similar results have been described by previous authors (Daeschel and Klaenhammer 1985;Gonzalez and Kunka 1987;Bhunia et al. 1988;Osmanagaoglu et al. 2001;Jamuna and Jeevaratnam 2004;Todorov and Dicks 2005). However, Elegado et al. (1997) found that the bacteriocin activity of pure pediocin at high concentrations remained even after incubation at pH 1AE0-12, although antimicrobial activity at a low bacteriocin concentration was detected at pH 1AE0-9AE0. ...
Screening and partial characterization of a bacteriocin produced by Pediococcus pentosaceus K23-2 isolated from Kimchi, a traditional Korean fermented vegetable.
A total of 1000 lactic acid bacteria were isolated from various Kimchi samples and screened for the production of bacteriocin. Pediocin K23-2, a bacteriocin produced by the Pediococcus pentosaceus K23-2 strain, showed strong inhibitory activity against Listeria monocytogenes. The bacteriocin activity remained unchanged after 15 min of heat treatment at 121 degrees C or exposure to organic solvents; however, it diminished after treatment with proteolytic enzymes. The bacteriocin was maximally produced at 37 degrees C, when the pH of the culture broth was maintained at 5.0 during the fermentation, although the optimum pH for growth was 7.0. The molecular weight of the bacteriocin was about 5 kDa according to a tricine SDS-PAGE analysis.
Pediococcus pentosaceus K23-2 isolated from Kimchi produces a bacteriocin, which shares similar characteristics to the Class IIa bacteriocins. The bacteriocin is heat stable and shows wide antimicrobial activity against Gram-positive bacteria, especially L. monocytogenes.
Pediocin K23-2 and pediocin K23-2-producing P. pentosaceus K23-2 could potentially be used in the food and feed industries as natural biopreservatives, and for probiotic application to humans or livestock.
... Considering previous successful applications of Ped. pentosaceus as a food preservative Jiang et al., 2021;Osmanagaoglu et al., 2001), Ped. pentosaceus LBM18 isolated from corn grain silage (Azevedo, Mendonça, Moreno, et al., 2020), which presented antibacterial and antifungal properties, was selected as a potential candidate to be used in the preservation of corn grain silage. ...
Aims:
This study aims to demonstrate the potential of the lactic acid bacteria (LAB) Pediococcus pentosaceus LBM18 against the mycotoxin-producing Alternaria alternata TEF-1A and highlight its application as an effective grain silage inoculant to control mycotoxin contamination.
Methods and results:
The antifungal properties of Ped. pentosaceus lyophilized (PPL) were assessed by evaluating its effect on A. alternata TEF-1A grown in a corn silage-based medium, which included morphological changes by Scanning Electron Microscopy (SEM) observations, growth rate, conidia production assays, and inhibition of Tenuazonic acid (TeA) production by high-performance liquid chromatography (HPLC-MS/MS) analyses. Furthermore, TeA biosynthesis was monitored for changes at the molecular level by PKS gene expression. The growth and sporulation processes of A. alternata TEF-1A were affected by Ped. pentosaceus LBM18 in a concentration-dependent manner. Moreover, a significant inhibition of TeA production (74.3%) and the transcription level of the PKS gene (42.9%) was observed.
Conclusions:
Ped. pentosaceus is one of the promising LAB to be applied as an inoculant for corn silage preservation, aiming to inhibit mycotoxigenic fungi growth and their mycotoxin production.
Significance and impact of the study:
Ped. pentosaceus could be used as an inoculant to reduce fungal and mycotoxins contamination in grain silage production.
... A higher temperature was selected as the optimum for pediocin production for P. acidilactici F (a sausage isolate) [69]. Production of pediocins from P. pentosaceus strains was at 37°C for P. pentosaceus ACCEL [70], and P. pentosaceus L and S (isolated from pork meat) [71], 35°C for P. pentosaceus Pep1 (isolated from sausages) [72], 30°C for P. pentosaceus ST18 (isolated from boza drink) [73] and P. pentosaceus SM-1 (isolated from pork sausage) [56]. Pediocin PD-1 from Pediococcus damnosus NCFB1832 [54] is produced at 30°C. ...
Class IIa bacteriocins from lactic acid bacteria are small, cationic proteins with antilisterial activity. Within this class, the pediocins are those bacteriocins that share a highly conserved hydrophilic and charged N-terminal part harboring the consensus sequence -YGNGV- and a more variable hydrophobic and/or amphiphilic C-terminal part. Several pediocins have been isolated and characterized. Despite the structural similarities, their molecular weight varies, as well as their spectrum of antimicrobial activity. They exhibit important technological properties, e.g. thermostability and retaining of activity at a wide pH range, which along with the bactericidal action against Gram-positive food spoilage and pathogenic bacteria, make them an important class of biopreservatives. Much new information regarding the pediocins has emerged during the last years. In this review, we summarize and discuss all the available information regarding the sources of pediocins, the characteristics of their biosynthesis and production in fermentation systems, the characteristics of the known pediocin molecules, and their antibacterial action. The advances made by genetic engineering in improving the features of pediocins are also discussed, as well as their perspectives for future applications.
... Gut microbiota of urban rats P. pentosaceus can be found in plant materials, ripe cheeses, and various processed meats (Osmanağaoğlu et al. 2001). It is also a starter culture capable of fermenting various meats, vegetables, and cheeses (Hu et al. 2007). ...
Mammals have a symbiotic relationship with various microorganisms called microbiota throughout their lives. These microorganisms are known to affect the host's physiology, health, and even mental balance. The development of the gut microbiota is regulated by a complex interaction between host and environmental factors, including diet and lifestyle. Herein, it is aimed to elucidate the changes in the gut microbiota of rats living in urban and rural habitats. All taxonomic changes in the gut microbiota of wild rats belonging to Rattus rattus species caught from urban and rural areas of Western Anatolian (Bilecik province) were examined comparatively by 16S rRNA next-generation sequencing technique. Laboratory rats were used as a control group for comparison. Thus, 2000 different bacterial species were identified in gut microbiota. According to the Shannon and Simpsons values calculated, laboratory rats showed the highest species diversity. When the similarities of microbiota profiles were compared with the principal coordinate analysis (PcoA), bacterial populations showed variability among different habitats. The comparison of species richness between the groups with the species rarefaction technique revealed higher species richness in all wild rats, especially in the rural habitat, compared to laboratory rats.
... Gut microbiota of urban rats Interestingly, 97% of P. pentosaceus was detected in the urban rats' group (Table 2). This bacterium can be found in plant materials, ripe cheeses, and various processed meats (Osmanağaoğlu et al. 2001). It is also known to be an industrially important bacterium because it is a starter culture capable of fermenting various meats, vegetables, and cheeses (Hu et al. 2007). ...
Mammals have a symbiotic relationship with various microorganisms called microbiota throughout their lives. These microorganisms are known to affect the host's physiology, health, and even mental balance. In the harbor of the densest and most diverse microorganisms in mammals, the curved structure of the intestines and their rich nutrient content are effective. The development of the gut microbiota is regulated by a complex interaction between host and environmental factors, including diet and lifestyle. Herein, it is aimed to elucidate the changes in the gut microbiota of rats living in urban and rural habitats. All taxonomic changes in the gut microbiota of wild rats belonging to Rattus rattus species caught from urban and rural areas of Western Anatolian (Bilecik province) were examined comparatively by 16S rRNA next-generation sequencing technique. Laboratory rats were used as a control group for comparison. Thus, 2000 different bacterial species were identified in gut microbiota. According to the Shannon and Simpsons values calculated, laboratory rats showed the highest species diversity. When the similarities of microbiota profiles were compared with the principal coordinate analysis (PcoA), bacterial populations showed variability among different habitats. The comparison of species richness between the groups with the species rarefaction technique revealed higher species richness in all wild rats, especially in the rural habitat, compared to laboratory rats. Food sources were determined as the most important factor contributing to species richness and diversity. While the increased food variety boosted species richness, species diversity was increased due to the diminished food variety.
... than 1% in all sample but one, is observed as fermentation proceeds. In this regard, some Pediococcus spp., which were more abundant at the end of the fermentation process, have shown the ability to inhibit several species of food pathogens (Osmanagaoglu et al., 2001). ...
Sesotho is an indigenous cereal-based fermented drink traditionally produced in the mountain kingdom of Lesotho, Southern Africa. The present study sought to examine the microbial (bacterial and fungal) community composition of Sesotho at five fermentation stages in five different locations. Using culture-independent (Illumina sequencing) techniques it was found that the bacterial communities followed similar successional patterns during the fermentation processes, regardless of geographical location and recipe variation between breweries. The most abundant bacterial taxa belonged to the phyla Firmicutes (66.2% of the reads on average) and Proteobacteria (22.1%); the families Lactobacillaceae (54.9%), Enterobacteriaceae (14.4%) and Leoconostrocaceae (8.1%); and the genera Lactobacillus (54%), Leuconostoc (10.7%), Leptotrichia (8.5%), and Weissella (5.5%). Most fungal taxa were from the phyla Ascomycota (60.7%) and Mucoromycota (25.3%); the families Rhizopodaceae (25.3%), Nectriaceae (24.2%), Saccharomycetaceae (16%) and Aspergillaceae (6.7%); and the genera Rhizopus (25.3%), Saccharomyces (9.6%), and Aspergillus (2.5%). Lactic acid bacteria (LAB) such as Enterococcus, Pediococcus, Lactobacillus, Leuconostoc, and Wiesella; as well as yeasts belonging to the genus Saccharomyces, were dominant in all breweries during the production of Sesotho. Several pathogenic and food spoilage microorganisms (e.g., Escherichia, Shigella, Klebsiella, etc.) were also present, but the study demonstrated the safety potential of the Sesotho fermentation process, as these microbial groups decline throughout Sesotho production. The functional profiles of the different brewing steps showed that the process is dominated by chemoheterotrophic and fermentative metabolisms. This study reveals, for the first time, the complex microbial dynamics that occur during Sesotho production.
... Several bacteriocins produced by LAB were reported to be stable at a pH ranging from 2 to 10 as BLIS from P. acidilactici P9 (Jia et al. 2014), BLIS from Lb. plantarum DM5 (Das and Goyal 2014), BLIS from Lb. plantarum BM-1 (Zhang et al. 2013) and BLIS from Lb. plantarum Lp6SH (Marie et al. 2012). BLIS activity was not affected by exposure to NaCl and organic solvents at a concentration of 50% (Table 4), which were similar for BLIS of P. pentosaceus K23-2 (Shin et al. 2008), pediocin of P. pentosaceus Pep1 (Osmanagaoglou et al. 2001) and BLIS of Bacillus cereus NS02 (Senbagam et al. 2013). The observations from above are in agreement with the previous reports by Kimura et al. (1997) which claimed that pediocin (bacteriocin from Pediococcus strains) is resistant to heat and some organic solvents and is active over a wide range of pH. ...
This paper deliberates the modelling and validation of bacteriocin-like inhibitory substance (BLIS) secretion by Pediococcus acidilactici Kp10 at different agitation speeds in a stirred tank bioreactor. A range of models namely the re-parameterised logistic, Luedeking–Piret and maintenance energy were assessed to predict the culture performance of the said bacterium. Growth of P. acidilactici Kp10 was enhanced with increased agitation speed up to 600 rpm while BLIS secretion was maximum at 400 rpm but decreased at higher agitation speed. Growth of P. acidilactici aptly subscribed to the re-parameterised logistic model while BLIS secretion and lactose consumption fitted well with the Luedeking–Piret model. The models revealed a relationship between growth of the bacterium and BLIS secretion. Bacterial growth and BLIS secretion were largely affected by the agitation speed of the stirred tank bioreactor which regulated the oxygen transfer to the culture. BLIS secretion by P. acidilactici Kp10 was however enhanced in oxygen-limited culture. The study also assessed BLIS from the perspective of its stability when subjected to factors such as temperature, pH and detergents. Results showed that BLIS produced by this strain was not affected by heat (at 25–100 °C for 20 min and at 121 °C for 15 min), surfactant (Tween 40, 60 and 80 and urea), detergents (up to 1% SDS), organic solvents (50% each of acetone, methanol and ethanol) and stable in a wide range of pH (2–10). The above information are pertinent with reference to commercial applications of this bacterial product in food manufacturing which invariably involve various sterilization processes and subjected to a wide pH range.
... Untuk produksi pediosin PaF-11,1 % inokulum P. acidilactici F-11 ditumbuhkan dalam media TGE cair 21,22 dengan volume 500 ml, pH 6,5 dan diinkubasi pada suhu 37 o C selama 18 jam. Purifikasi pediosin PaF-11 dilakukan dengan metode adsoprsi desorpsi yang dikembangkan Yang et al 23 . ...
Pediosin PaF-11 dari Pediococcus acidilactici F-11 merupakan peptida antibakteri yang aktif pada kisaran pH luas dan stabil pada perlakuan suhu tinggi dan rendah sehingga potensial digunakan sebagai pengawet tahu. Penelitian ditujukan untuk mengetahui mekanisme awal penghambatan pediosin PaF-11 sebagai antibakteri dan aplikasinya sebagai pengawet tahu. P. acidilactici F-11 dan Lactobacillus pentosus LB42 berturut turut digunakan sebagai bakteri penghasil dan indikator uji aktivitas pediosin PaF-11. Mekanisme awal kerja penghambatan pediosin PaF-11 ditentukan berdasar kajian pengaruh gadolinium (Gd3+) terhadap aktivitas pediosin PaF-11, kadar Gd3+ pada dinding sel dan morfologi sel indikator. Uji aktivitas pediosin PaF-11 dilakukan dengan metode difusi agar sumur. Aplikasi pediosin PaF-11 dan bakteriosin komersial nisin dilakukan terhadap tahu produksi CV. Kitagama Yogyakarta. Perlakuan meliputi perendaman tahu dalam larutan nisin (500 IU/g, 1000 IU/g dan 2000 IU/g) dan larutan pediosin PaF-11 30 AU/g. Setelah perendaman, dilanjutkan dengan pasteurisasi pada suhu 90oC selama 10 menit dan penyimpanan pada suhu 4oC selama 16 hari. Tahu tanpa perendaman dalam larutan nisin dan pediosin PaF-11 digunakan sebagai kontrol. Hasil penelitian mengindikasikan bahwa mekanisme awal penghambatan pediosin PaF-11 sebagai antibakteri yaitu melalui interaksi pediosin PaF-11 yang bermuatan positif dengan asam teikoat dan asam lipoteikoat yang bermuatan negatif. Hasil penelitian menunjukkan bahwa total bakteri awal pada tahu kontrol adalah 105 dan mengalami kenaikan menjadi 108 setelah disimpan. Penambahan larutan nisin dengan konsentrasi minimal 500 IU/g mampu menghambat populasi bakteri pada tahu sebesar 2 log cycle sedangkan larutan pediosin PaF-11(30 AU/g) mampu menghambat populasi bakteri pada tahu sebesar 0,5 log cycle.
... Being mild antimicrobials, pediocins are also expected in the future to find more applications in both human and veterinary medicine. Glucose, ribose, xylose, fructose and galactose to DL-lactate Sauerkraut Fermentations, dry sausages118119120121 Pediococcus pentosaceus Glucose, ribose, galactose, arabinose, and fructose to DL-lactate Starter cultures in sausage fermentations, brewing industry and, silage fermentations122123124 Pediococcus damnosus Glucose, sucrose and galactose Exopolysaccharide production and bacteriocin production [125,126] Lacticin 3147 Lacticin 3147 a class I, two-component bacteriocin produced by Lactococcus lactis subsp. lactis, isolated from an Irish kefir grain has been used for making buttermilk. ...
Bacteriocins are antimicrobial peptides which are ribosomally synthesized and produced by Lactic acid bacteria. They play a major role in prevention of human disease such as cancer, inflammatory disease, respiratory infection, systemic infection, intestinal disorder and bacterial infection and also contribute in maintaining the healthy gut microflora. Now day's bacteriocin is emerging as the very promising natural alternative against the antibiotic and chemical preservatives and gaining commercial importance worldwide. The inhibition of pathogenic bacterial strains occurs due to cell permiabilization, but producing strains are protected from it by specific immunity proteins. They are mainly classified in 4 classes: class I, class II, class III, and class IV bacteriocin based on lantibiotics ring. Nisin, Pediocin, Lactococcin B, Acidocin CH5, Curvacin A, and Sakacin are the bacteriocins, which have strong inhibition against pathogenic bacterial strain and used in food preservation. This review article summarizes and focuses on general introduction, classification, ecology and potential applications of bacteriocin as biopreservatives in food industry.
... There are several reports on the antilisterial effects of Pediococcus spp. isolated from Turkish fermented sucuk [4,23,24]. Also, some researchers [8,25] have investigated the behavior of L. monocytogenes during sucuk fermentation using bacteriocin-producer starter cultures. As far as we could determine, however, this is the first report on the antilisterial effects of a Pediococcus sp., isolated from a naturally fermented sucuk sample, during sucuk ripening. ...
This study evaluated control of Listeria monocytogenes during sucuk (a dry fermented sausage) ripening and storage of a sliced turkey breast product with Pediococcus acidilactici 13, which had been originally isolated from naturally fermented sucuk. When P. acidilactici 13 was used as a starter culture for sucuk production, L. monocytogenes counts decreased by 3.32 log CFU•g-1 during the 8-day ripening period, whereas the reduction in control samples was 1.37 log CFU•g-1 (P < 0.05). Treatment of turkey breast slices with partially purified substance of P. acidilactici 13 resulted in an immediate reduction by 1.03 log CFU•cm-2 (P < 0.05). It was concluded that P. acidilactici 13 could be useful as a protective culture for control of L. monocytogenes in particular in fermented meat products. The antimicrobial substance produced by this strain could only reduce the contamination by L. monocytogenes in a non-fermented meat product.
... Synthesis initially was detected in P. acidilactici strains (Bhunia et al., 1988;Gonzalez and Kunka, 1987) but P. pentosaceus has also ability to produce pediocin AcH as recently described by Bagenda et al. (2008) in a marine fermented seafood product. Other pediocins have been associated to P. pentosaceus, including pediocin A (Fleming et al., 1975), pediocin N5p (Strasser de Saad and Manca de Nadra, 1993), pediocin P (Osmanağaoğlu et al., 2001), pediocin ACCEL (Wu et al., 2004), pediocin SM-1 (Anastasiadou et al., in press). In recent years, there have been numerous reports on bacteriocin-producing enterococci, primarily among E. faecium associated with food systems (Giraffa, 2003). ...
... There are several reports on the antilisterial effects of Pediococcus spp. isolated from Turkish fermented sucuk [4,23,24]. Also, some researchers [8,25] have investigated the behavior of L. monocytogenes during sucuk fermentation using bacteriocin-producer starter cultures. As far as we could determine, however, this is the first report on the antilisterial effects of a Pediococcus sp., isolated from a naturally fermented sucuk sample, during sucuk ripening. ...
This study evaluated control of Listeria monocytogenes during sucuk (a dry fermented sausage) ripening and storage
of a sliced turkey breast product with Pediococcus acidilactici 13, which had been originally isolated from naturally
fermented sucuk. When P. acidilactici 13 was used as a starter culture for sucuk production, L. monocytogenes counts
decreased by 3.32 log CFU·g-1 during the 8-day ripening period, whereas the reduction in control samples was 1.37 log CFU·g-1 (P < 0.05). Treatment of turkey breast slices with partially purified substance of P. acidilactici 13 resulted in an immediate reduction by 1.03 log CFU·cm-2 (P < 0.05). It was concluded that P. acidilactici 13 could be useful as a protective culture for control of L. monocytogenes in particular in fermented meat products. The antimicrobial substance produced by this strain could only reduce the contamination by L. monocytogenes in a non-fermented meat product.
... There are several reports on the antilisterial effects of Pediococcus spp. isolated from Turkish fermented sucuk [4,23,24]. Also, some researchers [8,25] have investigated the behavior of L. monocytogenes during sucuk fermentation using bacteriocin-producer starter cultures. As far as we could determine, however, this is the first report on the antilisterial effects of a Pediococcus sp., isolated from a naturally fermented sucuk sample, during sucuk ripening. ...
Summary
This study evaluated control of Listeria monocytogenes during sucuk (a dry fermented sausage) ripening and storage
of a sliced turkey breast product with Pediococcus acidilactici 13, which had been originally isolated from naturally
fermented sucuk. When P. acidilactici 13 was used as a starter culture for sucuk production, L. monocytogenes counts
decreased by 3.32 log CFU·g-1 during the 8-day ripening period, whereas the reduction in control samples was
1.37 log CFU·g-1 (P < 0.05). Treatment of turkey breast slices with partially purified substance of P. acidilactici 13
resulted in an immediate reduction by 1.03 log CFU·cm-2 (P < 0.05). It was concluded that P. acidilactici 13 could be
useful as a protective culture for control of L. monocytogenes in particular in fermented meat products. The antimicrobial
substance produced by this strain could only reduce the contamination by L. monocytogenes in a non-fermented
meat product.
Keywords
Pediococcus acidilactici; Listeria monocytogenes; fermented sausage; turkey breast; protective culture
... The nucleotide sequences of the encoding gene showed 99% homology to that of pediocin operon of P. acidilactici strain K10 (GenBank accession number AY705375.1). Similar reports on the detection of pediocin have been observed in other strains of Pediococcus (Marugg et al. 1992;Osmanagaoglu et al. 2001;Millette et al. 2008). The pediocin maintained full activity at 70C for 20 min while loss of about 10% was observed on longer exposure (Table 3). ...
In an effort to control spoilage and pathogenic organisms in tsire , a Nigerian stick meat, the potential of a bacteriocinogenic Pediococcus acidilactici NCIMB 700993 as protective culture was evaluated. Preliminary experiments showed that P. acidilactici produced pediocin in an in vitro assay, among twelve strains of Pediococcus that were tested. Confirmation by polymerase chain reaction revealed that the organism encode gene for pediocin production of approximately 710 bp; its nucleotide sequences showed 99% homology to pediocin operon of P. acidilactici , (GenBank accession number AY705375.1). The pediocin exhibited full activity at 70C for 2 h while loss in activity was observed when treated with proteolytic enzymes, suggesting it may be proteinaceous in nature. Culture of P. acidilactici was inoculated on tsire at 6 log cfu/g and stored for 7 days at 30C to represent ambient temperature in Nigeria. Microbiological analysis showed reductions in the Enterobacteriaceae, coliforms, yeast & moulds, Staphylococcus and total bacterial counts in starter inoculated samples. In a challenge test, foodborne pathogens, Listeria monocytogenes and Salmonella Typhimurium, were initially inoculated at 6 log cfu/g on tsire samples; treatment with the P. acidilactici isolate reduced counts by 2 and 4 log, respectively, during storage while increase was observed in uninoculated control samples.
PRACTICAL APPLICATION
The present study focused on the potential of biological preservatives in the control of spoilage and pathogenic organisms in a traditional stick meat product tsire in Nigeria. Our findings may prove useful in the preservation of the product, as possible transformation into practical application could constitute an important economically viable approach for improving food safety and promoting public health. However, it would be imperative that the sellers of the product be given required training, especially on the need for observing personal hygiene and good manufacturing practices in order to reduce or prevent contamination of the product.
... Synthesis initially was detected in P. acidilactici strains (Bhunia et al., 1988; Gonzalez and Kunka, 1987) but P. pentosaceus has also ability to produce pediocin AcH as recently described by Bagenda et al. (2008) in a marine fermented seafood product. Other pediocins have been associated to P. pentosaceus, including pediocin A (Fleming et al., 1975), pediocin N5p (Strasser de Saad and Manca de Nadra, 1993), pediocin P (Osmanağaoğlu et al., 2001), pediocin ACCEL (Wu et al., 2004), pediocin SM-1 (Anastasiadou et al., in press). In recent years, there have been numerous reports on bacteriocin-producing enterococci, primarily among E. faecium associated with food systems (Giraffa, 2003). ...
This work had as main objectives to characterize two bacteriocins produced by lactic acid bacteria (LAB) previously isolated from non-fermented seafood, in order to evaluate their potential as new food protective agents. The two bacteriocinogenic isolates were identified by Polymerase Chain Reaction (PCR) using genus- and species-specific primers, and confirmed by 16S rDNA sequencing, as Enterococcus faecium and Pediococcus pentosaceus. The antimicrobial spectrum of each strain included several indicator microorganisms, some of them also isolated from seafood. Growth of Listeria innocua, L. monocytogenes, Staphylococcus aureus, Bacillus cereus and other LAB species were inhibited, although no inhibition of Gram-negative microorganisms was observed. Proteolytic, but not lipolytic or glycolytic enzymes, completely inactivated the antimicrobial effect of both cell-free supernatants confirming the proteinaceous nature of the inhibitors. The antimicrobial activity was maintained after treatment with NaCl, SDS, Triton X-100, Tween 20, Tween 80 and EDTA after 2 h or 5 h of exposure and both bacteriocins were stable over a wide range of pH and temperatures. Production of bacteriocin by E. faecium (bacALP7) was detected initially at exponential phase and reached a maximum activity of 25,600 AU/ml in the early stationary phase, whereas bacteriocin production by P. pentosaceus ALP57 (bacALP57) reached the maximum at exponential phase with 12,800 AU/ml. The bacteriocins did not kill L. monocytogenes ESB54 nor L. innocua 2030c however, cellular growth was reduced. The partially purified bacteriocins, bacALP7 and bacALP57, were below 6.5 kDa in size as determined by Tricine-SDS gel electrophoresis. E. faecium and P. pentosaceus contained DNA fragments corresponding in size to those recorded for enterocin B and pediocin PA-1, respectively. Sequencing of the fragments from both bacteriocins confirmed the homology. To our knowledge, for the first time two LAB producing bacteriocins similar to pediocin PA-1 and enterocin B, were isolated from non-fermented shellfish. The adaptation of the cultures to seafood matrices may be advantageous in terms of application as a biopreservation strategy for reduction of L. monocytogenes levels in seafood products.
With the aim of selectively isolating and understanding the cultivable lactic acid bacteria that are autochthonous to human gut, biopsy samples were collected from the terminal ileum and caecum of healthy volunteers who underwent routine colonoscopy procedure. The use of tissue samples (over stool samples) provides for a better expression of the autochthonous bacterial population of the human gut. The strains that were stable after many rounds of sub-culture were identified and studied further. One such ileal isolate, a Gram-positive, catalase-negative cocci was identified to be Pediococcus pentosaceus I44. It was studied for its gastric tolerance, bile tolerance and surface properties, and reported here. The effect of tween-80, oleic acid, extra virgin olive oil and sesame oil on the aforementioned properties of P. pentosaceus I44 was also analysed. The isolate was able to survive well in simulated gastric fluid of pH 3.0 followed by treatment in simulated intestinal fluid with a survival percentage of around 70%. It was, however, unable to withstand pH 2.0 even in the presence of supplements. It showed good tolerance to bile stress (1% w/v), and its cell surface was found to be hydrophilic. P. pentosaceus I44 showed good aggregation of 87% after 24h, with oleic acid having a significant effect on the isolate's aggregation potential. P. pentosaceus I44 is one of the few mucosa-bound cultivable bacteria that are adapted for survival in human colon. Studying and analysing such isolates might provide useful insights into their role in metabolism and health.
bacteriocin like inhibitory substance, BLIS, B. tenuis, antimicrobial activity, biopreservative Molecular weight of bacteriocin like substance (BL1S) of a new strain of Bacillus lentus 121 was found to be approximately 11 kDa. Purification of BLIS was attained by single step gel exclusion chromatography. BL1S was characterized by studying the inhibitory spectrum. It was active at broad pH range, high temperature and high NaCI concentration and showed sensitivity to proteolytic enzymes like trypsin, a-chymotrypsin and papain, the characters desirable for food preservation. BLIS extended the shelf stability of milk upto 21 days as a bio-preservative.
A total of 302 bacterial strains were isolated from 43 raw milk samples collected from dairy farms in Chiang Mai province. Out of these strains isolated, three strains were found to produce bacteriocins against Streptococcus dysgalactiae DMST10953, a bovine mastitis pathogen. These were Lactobacillus fermentum RMM701, Streptococcus bovis RMM703 and Streptococcus bovis RMM902, which exhibited bacteriocin activity at 40, 40 and 20 AU/ml respectively. Bacteriocins produced by these three strains were heat-stable up to 80 °C. Bacteriocins produced by L. fermentum RMM701 and S. bovis RMM902 were stable at pH 2.0 - 7.0 while that from S. bovis RMM703 was stable at pH 2.0 - 6.0. These bacteriocins were also found to be inactivated by proteolytic enzymes such as proteinase K and subtilisin A. © 2009 by Maejo University, San Sai, Chiang Mai, 50290 Thailand.
Rimbo Panti hot springs located bordering the province of West Sumatra and North Sumatra, precisely in the District of East Pasaman Pasaman regency of West Sumatra Province approximately 200km from the city of Padang. Hot water samples obtained from 5 pools which have a normal temperature of 50-95°C. The aim of this research was to isolate and identify Lactic Acid Bacteria (LAB) thermophilic activity from hot spring with pathogen bacteria (Escherichia coli 0157: H7, Salmonella thypmurium and Listeria monocytogenes) followed identified by PCR. The bacteria isolated were growth on medium thermus cair and then deMan Rogosa and Sharpe (MRS) Agar supplement with CaC03 1% and then performed purification by plate out on deMan Rogosa and Sharpe (MRS) Agar. It was found 23 isolates of LAB showed with clear zone around the culture and 5 isolates (N2, N4, N6, N9 and N12) has been antimicrobial activity against the growth of pathogenic bacteria. The results showed that isolates N6 had the highest antimicrobial activity against all bacteria test, with a range of inhibition zone 18-30 mm, gram positive, spore former coccus, non motility and catalase negative. LAB isolates that have the widest diameter of the clear zone continued to test the minimum inhibitory concentration (MIC). MIC values of isolates N6 supernatant against pathogenic bacteria Escherichia coli 0157: H7 by 60 and 80% of the bacterial pathogen Salmonella thypmurium and 50% of the bacterial pathogen Listeria monocytogenes. Based on morphological examination and PCR analysis, the isolate N6 was primarily identified as Pediococcus pentosaceus strain A24 bacteria.
The aim of this study was to isolate and identify anti-listerial substances produced by lactic acid bacteria in Kimchi, a traditional fermented vegetable. The Pediococcus pentosaceus T1 strain, which has an antimicrobial effect on Listeria monocytogenes, was isolated from Kimchi using 16S rRNA analysis. A crude culture of P. pentosaceus T1 demonstrated anti-listerial activity that was unaltered by pH changes in the range of 4–8 and temperatures between 80 and 110 °C. However, anti-listerial activity of P. pentosaceus T1 was abolished upon protease- and lipase-treatments, suggesting that the active substances were composed of peptides and lipids. Amylase, however, showed very little change in activity when compared to the control. Passage of the culture supernatant over Sep-Pak C18 cartridges showed that the anti-listerial activity could be traced to a component in the water-soluble fraction. Further purification of the activity was carried out using a series of steps that included ammonium sulfate precipitation, desalting, ion exchange chromatography, and ultrafiltration of the supernatants of P. pentosaceus cultures. The active fraction showed the presence of a 23-kDa protein, as visualized by SDS-PAGE followed by coomassie blue staining. Liquid chromatography (LC) and mass spectrometry (MS) analyses of the protein confirmed the presence of a Lysin motif (LysM) domain, which is known to be present in bacterial peptidoglycan hydrolases. In this study, we have demonstrated that Kimchi-derived P. pentosaceus shows an anti-listerial activity, and identified the active moiety as a LysM domain in a 23-kDa protein.
A Pediococcus acidilactici strain PA003 was isolated from traditionally fermented Chinese cabbage "Suan-tsai". The strain was found to produce an extracellular antimicrobial substance against Listeria monocytogenes and Lactobacillus plantarum. It was neither organic acid nor hydrogen peroxide. It was found sensitive to proteolytic enzymes and determined to be a proteinaceous bacteriocin in nature. The bacteriocin was resistant to heat and active over a wide range of pH. PCR amplification and the following cloning and sequencing of the amplified DNA fragment indicated that the bacteriocin was pediocin or its homolog.
All Gram-negative bacteria tested were normally resistant to the bactericidal action of pediocin P of Pediococcus pentosaceous Pep1, which had been isolated from vacuum-packaged sausages. However, when they were subjected to sublethal stress (exposure to physical and chemical stresses such as freezing, heating and acid treatment) and were treated with pediocin P, gram-negative bacterial cells that were normally resistant to pediocin P developed sensitivity to it and pediocin P reduced the viability of bacterial cells surviving sublethal stresses, although the extent of reduction varied with strain and type of treatment. When the combined effect of ethylenediaminetetraacetic acid (EDTA), sublethal heat and pediocin P was studied by adding EDTA and pediocin P to cell suspensions right before heating them, there was a considerable reduction in the number of viable cells, even at the lowest concentration of pediocin P tested. The results of our preliminary studies showed that sublethal injury can make the gram-negative bacterial cells sensitive to pediocin P.
Food-grade bacteria capable of producing bacteriocin with desirable preservation attri-butes have been isolated from traditional Indian fermented food dal vari, which has not been investigated so far. Among different isolates, Bacillus subtilis R75, isolated on MRS agar, exhibited antagonism against a wide range of foodborne pathogens that cause seri-ous spoilage. Extracellularly produced bacteriocin was purified by single step gel exclusion column chromatography. The purity rate and molecular mass of 12 kDa of this compound were determined using SDS-PAGE. Activity units (AU) of bacteriocin were increased in each step of purification, reaching up to 5·10 6 AU/mL. The increase in the activity units directly affected the antimicrobial activity of purified bacteriocin, resulting in an increase up to 200, 333 and 175 % of the inhibition zones against indicator bacteria. Continuous de-crease in the number of viable cells of microorganisms within 10 h after adding purified bacteriocin proved its bactericidal action. It withstood very high temperature, up to 121 °C, for 10 min, wider pH range, from 4.0 to 11.0, complete inactivation in the presence of pro-teolytic enzymes and storage stability up to 2.5 months.
This paper compares some important parameters and the free amino acid and biogenic amine contents of cured industrial and homemade meat products. To this aim, industrial and homemade "soppressata" and "salsiccia", typical dry fermented sausages produced in Southern Italy, were analyzed. The homemade sausages showed a higher level of free biogenic amines than that manufactured industrially, most likely because biogenic amine formation in industrial products is limited by the use of starter cultures. The industrial sausages are characterized by a higher total free amino acid content than the homemade products. Overall, free amino acid and biogenic amine contents demonstrated that appreciable differences exist between homemade and industrial sausages.
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