• Journal of Microbiology and Biotechnology
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• v.33 no.2
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• pp.211-218
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• 2023

A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Em^r), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.

• Microbiology and Biotechnology Letters
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• v.51 no.2
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• pp.147-156
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• 2023

Tetragenococcus halophilus CY54, an isolate from jeotgal, grows best in media with 5% NaCl and can grow at 18% and higher salt concentration. Three different doenjang samples were prepared with multiple starters including T. halophilus CY54. TBZA doenjang was prepared with T. halophilus, Bacillus subtilis, Zygosaccharomyces rouxii and Aspergillus oryzae. BZA doenjang was prepared with the same 3 starters except T. halophilus. KACC doenjang was prepared with a single starter, B. subtilis KACC16750. During 16 weeks of fermentation at 25℃, the viable counts were maintained in the range of 7-8 log CFU/g in all 3 samples. As fermentation progressed, pH decreased and titratable acidity (TA) gradually increased. Crude protein contents decreased slightly. TBZA doenjang showed higher amino-type nitrogen (ANN) and volatile basic nitrogen (VBN) contents, and KACC doenjang showed higher ammonia-type nitrogen (AMN) content. TBZA doenjang showed higher fibrinolytic and protease activity than other doenjang samples. Metabolites analyses by GC/MS showed that doenjang samples were separated from each other by partial least squares-discriminant analysis (PLS-DA) analysis. Seventeen major metabolites involved in the differences between samples were identified and they included organic acids, amino acids, sugars, fatty acids and alcohols. TBZA doenjang showed higher contents for most metabolites responsible for flavor and taste of fermented foods including doenjang. These results showed that T. halophilus could be useful as a starter for doenjang and can improve the product quality by accelerating the fermentation processes.

• Journal of Food Hygiene and Safety
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• v.38 no.4
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• pp.217-227
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• 2023

The consumption of ready-to-eat side dishes is rapidly growing in South Korea. These foods are particularly vulnerable to microbiological contamination as they are often cooked without any treatment, such as heating or stored at room temperature after cooking. Hence, in 2022, we analyzed the ready-to-eat side dishes sold in Gyeongsangnam-do, South Korea for microbiological contamination. We collected 100 samples from supermarkets in 7 cities, and then examined them for presence of food-borne pathogens and sanitary indicator bacteria. In the analysis of the food-borne pathogens, Bacillus cereus and Clostridium perfringens were isolated from 51 samples (51.0%) and 3 samples (3.0%), respectively. However, both quantitatively met the Korean Food Standards Codex. Genes of five different enterotoxins and one emetic toxin were analyzed from the 51 isolated B. cereus strains. We detected enterotoxin entFM (100.0%), nheA (94.1%), hblC (58.8%), cytK (56.9%), and bceT (41.2%) in 51 isolates, and emetic toxin gene, CER, in only one (2.0%) isolate. We did not detect C. perfringens toxin gene (cpe) that causes food poisoning in any one of the three C. perfringens isolates. In the case of sanitary indicator bacteria, Kimchi had the highest levels of total aerobic bacteria and coliforms, followed by Saengchae, Jeotgal, Jeolim, Namul, and Jorim, respectively. We counted total aerobic bacteria at two different storage temperatures (4℃ and 20℃) to determine the effect of storage temperature. When stored at 20℃, total aerobic bacteria count increased in most of the ready-to-eat side dishes, except for Jeotgal. This result conclusively shows the need for refrigerating the ready-to-eat side dishes after purchase. Further research is needed to assess the risk and safety of the ready-to-eat side dishes available in the market and determine appropriate safety management practices.

• Korean Journal of Food Science and Technology
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• v.45 no.5
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• pp.571-577
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• 2013

Our aim was to ferment soymilk using lactic acid bacteria that showed protease activity and to optimize the condition for fermentation. In total, 108 strains of protease-producing lactic acid bacteria were isolated from various fermented foods such as kimchi and jeotgal, and among them, 29 strains displaying the highest protease activity were selected for further study. From these 29 strains, strain MK1, whose protease activity was 126 $mU/mL{\cdot}min$, was selected as the optimal fermentation strain owing to its high ability to digest soymilk protein. It was henceforth labeled as Lactobacillus paracasei MK1. The optimum conditions for the fermentation of soymilk by using L. paracasei MK1 were determined to be as follows: 30 h of fermentation time at a temperature of $30^{\circ}C$, and at a pH of 6.0 in the initial growth medium.

• Korean Journal of Community Nutrition
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• v.17 no.5
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• pp.555-578
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• 2012

This study was conducted to investigate which food items are perceived as HANSIK (Korean food) in Korea. 562 males and females aged 20-70 were surveyed on 512 most frequently consumed dish items from 4th Korean National Health and Nutrition Survey if they perceive it as HANSIK. Dish items in kimchi, namul sukchae (cooked and seasoned vegetable) and jeotgal (salt-fermented food) category showed high response rate to be perceived as HANSIK. The response rate of twigim (frying foods) as HANSIK was low showing less than 70%. The response rate as HANSIK for foreign origin foods such as ramen and jjajangmyeon (black bean paste noodle) were lower than 25%. In jang yangnyum (seasoning) category, doenjang (soybean paste) and gochujang (red pepper paste) showed high response rate as HANSIK more than 90%. Females showed a higher response rate as HANSIK than males for most food items except several items of myen mandu (noodle and dumpling). The younger age group had higher HANSIK perception on many items of recently consumed foods, especially budae-jjigae (spicy sausage stew) (p < 0.001) and jwipo-jorim (braised dried filefish) (p < 0.001), implying that they already accept the common foods as HANSIK regardless of the origin. These results provide an information on the foods perceived as HANSIK among currently consumed common foods in Korea and these results can be utilized for establishment of HANSIK concept reflecting transition of dietary life in Korea.

• Journal of Species Research
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• v.6 no.2
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• pp.101-118
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• 2017

In an investigation of indigenous prokaryotic species in Korea, a total of 57 bacterial strains assigned to the classes Betaproteobacteria and Gammaproteobacteria were isolated from diverse environments. Samples were collected from fresh water, natural caves, soil, paddy fields, lakes, sea water, jeotgal (fermented seafood), salt flats, soil from abandoned mines, plant roots, digestive organs of both Japanese crested ibis (Nipponia nippon) and Burmese python (Python molurus bivittatus) and tidal flats. From the high 16S rRNA gene sequence similarity (>98.7%) and formation of robust phylogenetic clades with closely related species, it was determined that each strain belonged to an independent and predefined bacterial species within either the Betaproteobacteria or Gammaproteobacteria. There is no official report or publication that describes these 57 proteobacterial species in Korea. Overall, in the class Betaproteobacteria there were 16 species in 12 genera of 4 families in the order Burkholderiales and two species in two genera of one family in the order Neisseriales. Within the class Gammaproteobacteia, there were five species in four genera of four families in the order Alteromonadales, 12 species in 11 genera of one family in the order Enterobacteriales, four species in four genera of three families in the order Oceanospirillales, 11 species in four genera of two families in the order Pseudomonadales, two species in the order Vibrionales and five species in five genera of one family in the order Xanthomonadales. Gram reaction, colony and cell morphology, basic biochemical characteristics, isolation source and strain IDs are described in the species description section.

• Journal of Dairy Science and Biotechnology
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• v.31 no.2
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• pp.153-159
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• 2013

Lactic acid bacteria are microorganisms that are closely associated with human and/or animal environments, and are categorized as generally recognized as safe (GRAS) organisms due to their ubiquitous appearance in foods and their contribution to the healthy microflora of mucosal surfaces. This study was performed to isolate and identify lactic acid bacteria with antagonistic effects against food-borne pathogens. A total of 3,000 acid-producing bacteria were isolated from infant feces, cattle feces, goat feces, dog feces, pig feces, vaginal tracts, vegetables, fruits, Kimchi, Jeotgal, fermented sausages, raw milk, cheese, yogurt, Cheonggukjang, Meju, and Makgeolli cultured on MRS agar with 0.05% bromocresol purple. For the isolation of bacteriocin-producing bacteria, the diameter of the clear zone was measured on MRS agar plates. Twenty-six isolates exhibited strong antibacterial activity against indicator strains such as Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica serovar Enteritidis. Lactic acid bacteria were identified as Enterococcus faecalis, Enterococcus faecium, Enterococcus hirae, Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus curvatus, Lactobacillus plantarum, and Pediococcus acidilactici by 16S rDNA gene sequence analysis. The results of this study suggest that the isolates could be used as potential probiotic starters for functional food applications.

• Korean Journal of Microbiology
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• v.45 no.2
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• pp.193-199
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• 2009

A bacterium producing the halotolerant extracellular protease was isolated from squid jeotgal, and was identified as Bacillus sp. SJ-10 based on morphological, physiological and biochemical characteristics, as well as phylogenetic analysis using 16S rRNA gene sequence. The strain grew at $20^{\circ}C\sim55^{\circ}C$, pH 5~8, and 0%~14% NaCl and optimal growth conditions were $35{\pm}5^{\circ}C$, pH 7, and 5% NaCl. The major cellular fatty acids were anteiso-$C_{15:0}$, anteiso-$C_{17:0}$, and $C_{16:0}$ DNA G+C content was 50.58 mol% and menaquinone consisted of MK-7 Phylogenic analysis based on the 16S rRNA gene sequence indicated that SJ-10T belongs to the genus Bacillus. About 40 kDa of the salt-tolerant protease was purified by 40% ammonium sulfate saturation and Mono Q column chromatography. The optimal activity of the protease was pH 8 and stable at pH 5~10. The optimum temperature and NaCl concentration were $35{\pm}5^{\circ}C$ and $5{\pm}1%$, respectively.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.263-268
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• 2004

Two species of halophilic bacteria were isolated from five salted seafoods and identified by 16S rDNA sequenc­ing homology. One was identified as Halomonas subglaciescola and other four strains were belong to Halomo­nas marina. The identity of all isolates with standard organisms was above $95\%.$ H. subglaciescola, H. marina IN, and H. marina SH-2 grew in salinity condition from $3%\;to\;18\%$ NaCl but growth of H. marina SQ and H. marina SH-l grew in salinity environment from $8\%\;to\;17\%.$ Maximum biomass of H. subglaciescola, H. marina IN, H. marina SQ, H. marina SH-1, and H. marina SH-2 growing in LB medium containing $15\%$ NaCl were about 3.2, 4.5, 4.5, 5.7, and 4.2, however the maximum biomass in LB medium containing $5\%$ NaCl were about 2.2, 1.1, 0.7, 0.2, and 2.4 as optical density at 660 nm, respectively. In scanning electron micrograph, unknown material (mucus) attached to outer membrane of all isolates was observed. When mucus isolated from halophilic bacterial cell was added to culture of E. coli, E. coli grew in medium containing $15\%$ NaCl.

• Food Engineering Progress
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• v.21 no.1
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• pp.88-92
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• 2017

We attempted to investigate antibacterial and proteolytic activities of bacteria isolated from three ethnic fermented seafoods in the east coast of South Korea, gajami sikhae, squid jeotgal, and fermented jinuari (Grateloupia filicina). Bacillus cereus ATCC 14579, Listeria monocytogenes ATCC 15313, Staphylococcus aureus KCTC 1916, Escherichia coli O157:H7 ATCC 43895, and Salmonella enterica serovar Typhimurium ATCC 4931 were selected to determine the antibacterial activity of the bacterial isolates. Among 233 isolates from the three foods, 36 isolates (15.5%) showed antibacterial activity against B. cereus ATCC 14579, the highest incidence of inhibition, followed by S. aureus KCTC 1916 (7.7%) and L. monocytogenes ATCC 15313 (6.0%). However, only five and three strains among the isolates exhibited inhibitory activity against Gram-negative indicators, E. coli ATCC 43895 and Sal. enterica ATCC 4931, respectively. The proteolytic activity of the isolates was determined via hydrolysis of skim milk after 24, 48, and 72 h incubation. After 72 h incubation, 72 out of 233 isolates (30.9%) showed proteolytic activity, and the isolates of fermented jinuari exhibited the highest incidence of proteolytic activity (60%, 36 isolates). These results suggest that ethnic fermented seafoods in the east coast of South Korea might be a promising source of bacterial strains producing antibacterial and proteolytic compounds.