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Integrative genome analysis of bacteriocin-producing Lactiplantibacillus pentosus LNP1-39 and its synbiotic role in suppressing food-borne pathogens
(2026-06-01)
Jirakanjanasit, Thanadol
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Choovet, Natladda
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Booncharoen, Auttaporn
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Kingkaew, Engkarat
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Poothong, Saranporn
Lactic acid bacteria were isolated from traditional Thai-fermented foods. Among these, the strain LNP1-39, closely related to Lactiplantibacillus pentosus, was selected for further study because of its non-pathogenic profile. The bacteriocins produced by L. pentosus LNP1-39 were proteinaceous substances that exhibited strong antimicrobial activity across a wide pH range (pH 2–11; 6400–2400 AU/mL) and thermal stability at 100 °C for 40 min (400 AU/mL). These bacteriocins showed a narrow antimicrobial spectrum, effectively targeting Gram-positive pathogens, such as Kocuria rhizophila MIII, Enterococcus faecalis JCM 5803 T, and Listeria monocytogenes ATCC 19115. Comprehensive safety assessments, including whole-genome analysis and in vitro tests, confirmed a low risk of antibiotic resistance and the absence of virulence factors. Strain LNP1-39 was confirmed to be closely related to L. pentosus DSM 20314 T via digital DNA‒DNA hybridization (dDDH; 75.4%), with average nucleotide identity (ANI) at 96.56% ANIb and 97.22% ANIm values. Additionally, LNP1-39 produces pediocin with notable similarity (76.29% identity to pediocin) and presents low risks for antibiotic-resistance genes or transfer genes while providing antioxidant properties. Strain LNP1-39 survived harsh gastrointestinal tract conditions and exhibited a favorable prebiotic index and positive prebiotic activity score when paired with polydextrose or isomalto-oligosaccharide. These findings support L. pentosus LNP1-39 as potential bacteriocin-producing lactic acid bacteria for further application in food preservation and pathogen control or as a synbiotic.
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Functional and genomic characterization of Pediococcus pentosaceus NR4-4 reveals a safe bacteriocinogenic probiotic with strong anti-listerial activity and prebiotic metabolism
(2026-05-01)
Ratthanachot, Natnarin
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Woraprayote, Weerapong
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Janyaphisan, Thitiphorn
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Tanasupawat, Somboon
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Ochaikul, Duangjai
The growing interest in bacteriocin-producing lactic acid bacteria from traditional fermented foods has highlighted the value of strains that combine antimicrobial activity, probiotic functionality, and genome-supported safety. This study presents an integrated functional and probiogenomic characterization of Pediococcus pentosaceus NR4-4 isolated from Thai fermented mustard greens. The strain was found to produce a heat-stable class IIa bacteriocin with pronounced inhibitory activity against Listeria monocytogenes . In addition, NR4-4 exhibited strong tolerance to simulated gastrointestinal conditions and demonstrated effective adhesion to Caco-2 intestinal epithelial cells. In vitro safety evaluations confirmed that the strain was non-hemolytic and did not exhibit cytotoxic effects toward AGS or Caco-2 cell lines. Whole-genome sequencing verified species identity and revealed the presence of pediocin-related gene clusters, carbohydrate-active enzymes associated with fructooligosaccharide utilization, and a favorable genome-based safety profile. Notably, no virulence factors, biogenic amine biosynthesis pathways, plasmids, or transferable antimicrobial resistance determinants were detected. The preferential metabolism of fructooligosaccharides further suggests compatibility with synbiotic formulations. Overall, these findings indicate that P. pentosaceus NR4-4 is a genomically supported, safe, and multifunctional probiotic candidate with potential applications in food biopreservation and functional food development.
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Genomic Assessment, Metabolic Profile Mapping, and Anti-Helicobacter pylori Activity of Lactococcus lactis SK2-659 from Thai Fermented Green Mustard (Pak-kad-dong)
(2026-01-01)
Kingkaew, Engkarat
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Woraprayote, Weerapong
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Sitdhipol, Jaruwan
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Vilaichone, Ratha Korn
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Visessanguan, Wonnop
Probiotics play crucial roles in promoting gut health, enhancing immunity, and combating pathogenic microorganisms, with increasing interest in their applications in the food and therapeutic industries. Lactococcus lactis, a well-known lactic acid bacterium, has emerged as a promising candidate owing to its probiotic traits and safety profile. In this study, we investigated the probiotic potential and genomic profile of Lactococcus lactis SK2-659, a strain isolated from Thai fermented green mustard (Pak-kad-dong). Genomic analysis revealed numerous genes associated with probiotic traits, including stress tolerance, adhesion, and antimicrobial activity, with a particular focus on bacteriocin SK2-659, which was effective against pathogenic bacteria such as Helicobacter pylori. The bacteriocin produced by L. lactis SK2-659, identified as nisin Z, disrupts bacterial membranes via pore formation, leading to cell lysis. Metabolomic profiling further highlighted its ability to increase carbohydrate and amino acid metabolism, supporting cell growth and survival in acidic environments. Also, amino acid metabolism (elevated tryptophan, tyrosine, histidine) supports acid tolerance and immune modulation. Tryptophan metabolism produces indole derivatives, which are known to benefit gut health and immunity. Additionally, strain SK2-659 demonstrated strong tolerance to gastrointestinal conditions, high adhesion capacity to intestinal cells, and immunomodulatory effects, contributing to gut health. Safety assessments confirmed the absence of virulence factors, pathogenic traits, and antibiotic resistance genes, along with a lack of biogenic amine production, underscoring its suitability for food and health applications. These findings establish L. lactis SK2-659 as a promising probiotic candidate with potential industrial applications in functional foods, food preservation, and therapeutic products. Given its probiotic properties, antimicrobial activity, and safety profile, this strain is particularly suited for the food industry as a functional food ingredient and natural bio-preservative, as well as the healthcare and pharmaceutical industries for use in therapeutic probiotics and gut health formulations.
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Screening of bacteriocin-producing bacteria associated in traditional Thai fermented beef (MUM)
(2023-08-28)
Piayura, S.
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Pinsirodom, P.
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Surapantapisit, Y.
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Swetwiwathana, A.
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Effect of garlic on lactic acid and bacteriocins production of bacteriocin-producing lactic acid bacteria associated in Nham (Thai fermented meat) during fermentation
(2023-08-28)
Swetwiwathana, A.
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Lotong, N.
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Nakayama, J.
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Sonomoto, K.
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Effect of extracted pectin from fruit wastes on growth of Pediococcus pentosaceus RSU-Nh1 and Lactobacillus plantarum RSU-SO2
(2020-01-01)
Showpanish, K.
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Sonhom, N.
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Pilasombut, K.
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Woraprayote, W.
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Prachom, N.
Two strains of lactic acid bacteria identified as Pediococcus pentosaceus RSU-Nh1 and Lactobacillus plantarum RSU-SO2 were isolated from Thai fermented pork sausage (Nham) and fermented spring onion (Ton-Hom-Dong) respectively. The culture supernatant (CFS) of both strains exhibited antimicrobial activity against both Gram-positive and Gramnegative pathogenic bacteria. After being treated with alpha-chymotrypsin and trypsin, the antimicrobial activity of their CFS was eliminated, which suggests that the antimicrobial compound produced was antimicrobial peptide or bacteriocin. Pectin extracted from different fruit peels was supplemented in the bacterial growth medium to investigate the effect of pectin on the growth and antimicrobial production of Ped. pentosaceus RSU-Nh1 and Lb. plantarum RSU-SO2. Pectin was extracted from pomelo, durian, passion fruit and salacca fruit peels with a pectin yield of 5.02 to 26.44%, degree of esterification of 80.84 to 86.68%, and galacturonic acid content of 52.36 to 64.23%. Supplementation with pomelo and passion fruit pectin exhibited the highest improvement of bacterial growth and antimicrobial production of Ped. pentosaceus RSU-Nh1 and Lb. plantarum RSU-SO2. These findings suggested the potential application of these pectin extracts as prebiotics to promote the growth and antimicrobial production of lactic acid bacteria.
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Potential of bacteriocin-producing lactic acid bacteria for safety improvements of traditional Thai fermented meat and human health
(2015-11-01)
Swetwiwathana, Adisorn
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Visessanguan, Wonnop
Lactic acid bacteria (LAB) are very important in converting of agricultural products into safe, delicious and shelf stable foods for human consumption. The preservative activity of LAB in foods is mainly attributed to the production of anti-microbial metabolites such as organic acids and bacteriocins which enables them to grow and control the growth of pathogens and spoilage microorganisms. Besides ensuring safety, bacteriocin-producing LAB with their probiotic potentials could also be emerging as a means to develop functional meat products with desirable health benefits. Nevertheless, to be qualified as a candidate probiotic culture, other prerequisite probiotic properties of bacteriocin-producing LAB have to be assessed according to regulatory guidelines for probiotics. Nham is an indigenous fermented sausage of Thailand that has gained popularity and acceptance among Thais. Since Nham is made from raw meat and is usually consumed without cooking, risks due to undesirable microorganisms such as Salmonella spp., Staphylococcus aureus, and Listeria monocytogenes, are frequently observed. With an ultimate goal to produce safer and healthier product, our research attempts on the development of a variety of new Nham products are discussed.
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In vitro characterization of bacteriocin produced by lactic acid bacteria isolated from Nem Chua, a traditional Vietnamese fermented pork
(2015-08-01)
Pilasombut, Komkhae
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Rumjuankiat, Kittaporn
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Ngamyeesoon, Nualphan
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Duy, Le Nguyen Doan
The aim of this study was to screen and In vitro characterize the properties of bacteriocin produced by lactic acid bacteria isolated from Vietnamese fermented pork (Nem chua). One hundred and fifty LAB were isolated from ten samples of Nem chua and screened for bacteriocin-producing lactic acid bacteria. Antimicrobial activity of bacteriocin was carried out by spot on lawn method against both gram positive and gram negative bacteria. One isolate, assigned as KL-1, produced bacteriocin and showed inhibitory activity against Lactobacillus sakei, Leuconostoc mesenteroides and Enterococcus faecalis. To characterize the bacteriocin-producing strain, optimum temperature, incubation period for maximum bacteriocin production and identification of bacteriocin-producing strain were determined. It was found that the optimum cultivation temperature of the strain to produce the maximum bacteriocin activity (12,800 AU/mL) was obtained at 30°C. Meanwhile, bacteriocin production at 6,400 AU/mL was found when culturing the strain at 37°C and 42°C. The isolate KL-1 was identified as L. plantarum. Antimicrobial activity of cell-free supernatant was completely inhibited by proteolytic enzyme of trypsin, alpha-chymotrypsin and proteinase K. Bacteriocin activity was stable at high temperature up to 100°C for 10 min and at 4°C storage for 2 d. However, the longer heating at 100°C and 4°C storage, its activity was reduced.
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Purification and characterization of a novel plantaricin, KL-1Y, from Lactobacillus plantarum KL-1
(2015-06-13)
Rumjuankiat, Kittaporn
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Perez, Rodney Horanda
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Pilasombut, Komkhae
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Keawsompong, Suttipun
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Zendo, Takeshi
Three bacteriocins from Lactobacillus plantarum KL-1 were successfully purified using ammonium sulfate precipitation, cation-exchange chromatography and reverse-phase HPLC. The bacteriocin peptides KL-1X, -1Y and -1Z had molecular masses of 3053.82, 3498.16 and 3533.16 Da, respectively. All three peptides were stable at pH 2–12 and 25 °C and at high temperatures of 80 and 100 °C for 30 min and 121 °C for 15 min. However, they differed in their susceptibility to proteolytic enzymes and their inhibition spectra. KL-1Y showed broad inhibitory activities against Gram-positive and Gram-negative bacteria, including Salmonella enterica serovar Enteritidis DMST 17368, Pseudomonas aeruginosa ATCC 15442, P. aeruginosa ATCC 9027, Escherichia coli O157:H7 and E. coli ATCC 8739. KL-1X and -1Z inhibited only Gram-positive bacteria. KL-1X, KL-1Y and KL-1Z exhibited synergistic activity. The successful amino acid sequencing of KL-1Y had a hydrophobicity of approximately 30 % and no cysteine residues suggested its novelty, and it was designated “plantaricin KL-1Y”. Plantaricin KL-1Y exhibited bactericidal activity against Bacillus cereus JCM 2152T. Compared to nisin, KL-1Y displayed broad inhibitory activities of 200, 800, 1600, 800, 400 and 400 AU/mL against the growth of Bacillus coagulans JCM 2257T, B. cereus JCM 2152T, Listeria innocua ATCC 33090T, Staphylococcus aureus TISTR 118, E. coli O157:H7 and E. coli ATCC 8739, respectively, whereas nisin had similar activities against only B. coagulans JCM 2257T and B. cereus JCM 2152T. Therefore, the novel plantaricin KL-1Y is a promising antimicrobial substance for food safety uses in the future.
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Purification and amino acid sequence of a bacteriocins produced by Lactobacillus salivarius K7 isolated from chicken intestine
(2006-01-01)
Pilasombut, Komkhae
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Sakpuaram, Thavajchai
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Wajjwalku, Worawidh
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Nitisinprasert, Sunee
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Swetwiwathana, Adisorn
A bacteriocin-producing strain, Lactobacillus K7, was isolated from a chicken intestine. The inhibitory activity was determined by spot-on-lawn technique. Identification of the strain was performed by morphological, biochemical (API 50 CH kit) and molecular genetic (16S rDNA) basis. Bacteriocin purification processes were carried out by amberlite adsorption, cation exchange and reverse-phase high perform ance liquid chromatography. N-terminal amino acid sequences were performed by Edman degradation. Molecular mass was determined by electrospray-ionization (ESI) mass spectrometry (MS). Lactobacillus K7 showed inhibitory activity against Lactobacillus sakei subsp. sakei JCM 1157T, Leuconostoc mesenteroides subsp. mesenteroides JCM 6124T and Bacillus coagulons JCM 2257T. This strain was identified as Lb. salivarius. The antimicrobial substance was destroyed by proteolytic enzymes, indicating its proteinaceous structure designated as a bacteriocin type. The purification of bacteriocin by amberlite adsorption, cation exchange, and reverse-phase chromatography resulted in only one single active peak, which was designated FK22. Molecular weight of this fraction was 4331.70 Da. By amino acid sequence, this peptide was homology to Abp 118 beta produced by Lb. salivarius UCC118. In addition, Lb. salivarius UCC118 produced 2-peptide bacteriocin, which was Abp 118 alpha and beta. Based on the partial amino acid sequences of Abp 118 beta, specific primers were designed from nucleotide sequences according to data from GenBank. The result showed that the deduced peptide was high homology to 2-peptide bacteriocin, Abp 118 alpha and beta.

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