• 한국미생물·생명공학회지
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• 제47권4호
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• pp.522-529
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• 2019

A Bacillus strain, SJ4, exhibiting strong fibrinolytic activity was isolated from saeu (shrimp, Acetes chinensis) jeotgal, a Korean traditional fermented food and was identified as B. subtilis. The B. subtilis SJ4 strain can grow at a NaCl concentration of up to 15% (w/v). The fibrinolytic activity of B. subtilis SJ4 (152.0 U/ml) cultured in Luria-Bertani (LB) broth for 48 h at 37℃ with aeration was higher than that of B. subtilis SJ4 cultured in TSB (124.5 U/ml) under same culture conditions. The major proteins in the LB culture supernatant of B. subtilis SJ4 were analyzed by SDS-PAGE, which revealed three major bands (23, 25, and 28 kDa). The band (23 kDa) with strong fibrinolytic activity, analyzed on fibrin zymogram, was observed at 60-96 h of cultivation. The aprESJ4 gene encoding the major fibrinolytic enzyme, AprESJ4, was cloned by PCR. The aprESJ4 gene sequence exhibited high similarities with the fibrinolytic gene sequences of other Bacillus species. The amino acid sequence of AprESJ4 exhibited 98.9 and 98.4% similarity with subtilisin NAT and AprE2 of B. subtilis, respectively. Hence, B. subtilis SJ4 can be a potential starter culture for jeotgal products.

• 한국수산과학회지
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• 제54권6호
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• pp.1045-1051
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• 2021

Eighty-nine different types of commercially salt-fermented fishery products comprising various raw materials were analyzed for total aerobic bacteria, number of coliform bacteria, fecal coliform, and Escherichia coli. The food-poisoning bacterial content of the samples was investigated using next-generation sequencing. The mean mass of total aerobic bacteria in Jeotgal was 6-1.8×10⁹ CFU/g, and that in Aekjeot and Sikhae was 4-2.2×10⁵ CFU/mL and 1.9×10⁵-8.4×10⁸ CFU/g, respectively. Coliform bacteria were detected in 9 (28.1%) of 32 Jeotgal samples; 15 (46.8%) of 32 seasonal Jeotgal samples; and in 5 (55.5%) of 9 Sikhae samples. Fecal coliform and E. coli were not detected in 86 of the 89 samples. Yersinia enterocolitica was detected only in Galchi jeot (salt-fermented hairtail) (1 type) and not in other Jeotgal samples. These results contribute to our knowledge regarding the bacterial stability of salt-fermented fishery products.

• 한국미생물·생명공학회지
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• 제46권4호
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• pp.313-319
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• 2018

Halophilic lactic acid bacteria (LAB) were isolated from myeolchi-jeotgal (23% NaCl, w/v) fermented in jangdok (Korean earthenware) located outside a house. They were identified as Tetragenococcus halophilus by 16S rRNA and recA gene sequencing. Four T. halophilus isolates showing high protease activities were selected for further studies. Four strains grew well, reaching $OD_{600}$ values of 0.75-0.92 at 18% NaCl content (w/v) and 0.28-0.44 at 23% salt. They showed rapid growth, attaining $OD_{600}$ values of 1.1-1.2 at $20-30^{\circ}C$, but did not grow at $4^{\circ}C$. At $15^{\circ}C$, the highest $OD_{600}$ values, which exceeded 0.6, were observed at 20 days, and were higher than those of cultures at $37^{\circ}C$ and $42^{\circ}C$ (approximately 0.5). Four isolates grew best in broth where the initial pH was adjusted to 8 and did not grow at $pH{\leq}4$. T. halophilus BS2-36 showed the highest survival ratio of 18.7% after 2 hours of exposure at pH 3. BS2-36 showed the highest survival ratio (1.29%) in presence of 0.3% bile salts. T. halophilus BS2-36 seems a promising candidate as a starter for jeotgal and other fermented foods with high salinities.

• Journal of Microbiology and Biotechnology
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• 제29권3호
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• pp.347-356
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• 2019

Bacillus sp. BS2 showing strong fibrinolytic activity was isolated from sea squirt (munggae) jeotgal, a traditional Korean fermented seafood. BS2 was identified as B. velezensis by molecular biological methods. B. velezensis BS2 grows well at 15% NaCl and at $10^{\circ}C$. When B. velezensis BS2 was cultivated in TSB broth for 96 h at $37^{\circ}C$, the culture showed the highest fibrinolytic activity ($131.15mU/{\mu}l$) at 96 h. Three bands of 27, 35 and 60 kDa were observed from culture supernatant by SDS-PAGE, and fibrin zymography showed that the major fibrinolytic protein was the 27 kDa band. The gene (aprEBS2) encoding the major fibrinolytic protein was cloned, and overexpressed in heterologous hosts, B. subtilis WB600 and E. coli BL21 (DE3). B. subtilis transformant showed 1.5-fold higher fibrinolytic activity than B. velezensis BS2. Overproduced AprEBS2 in E. coli was purified by affinity chromatography. The optimum pH and temperature were pH 8.0 and $37^{\circ}C$, respectively. $K_m$ and $V_{max}$ were 0.15 mM and $39.68{\mu}M/l/min$, respectively, when N-succinyl-Ala-Ala-Pro-Phe-pNA was used as the substrate. AprEBS2 has strong ${\alpha}$-fibrinogenase and moderate ${\beta}$-fibrinogenase activity. Considering its high fibrinolytic activity, significant salt tolerance, and ability to grow at $10^{\circ}C$, B. velezensis BS2 can be used as a starter for jeotgal.

• 한국수산과학회지
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• 제57권4호
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• pp.489-496
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• 2024

This study has investigated ten different organic acids in 67 types of seasoning Jeotgal (seasoning salted fermented product, SSFP) and Sikhae (seasoning salted fermented product with rice, SSFPR) collected from the market. The levels of organic acids varied significantly depending on the raw materials used for the SSFP and SSFPR. The acetic acid and lactic acid content ranged from being not detected (ND) to 5,667.15 mg/kg for acetic acid in the SSFP, ND to 34,837.89 mg/kg for lactic acid in the SSFP, and 53.47 to 2,089.55 mg/kg for acetic acid in the SSFPR, and 10.69 to 1,733.27 mg/kg for lactic acid in the SSFPR. The range for succinic acid was 57.92 to 2,793.09 mg/kg in the SSFP and 21.21 to 453.32 mg/kg in the SSFPR. Propionic acid content ranged from 8.42 to 277.83 mg/kg in the SSFP and 13.21 to 133.07 mg/kg in SSFPR.The most abundant organic acids were succinic and lactic acid, comprising 48% of the seasoning Jeotgal (succinic acid) and more than 72% in Sikhae (lactic acid). Furthermore, this study suggests that the differences in the organic acid content of samples, based on their raw materials can be discriminated by principal component analysis (PCA).

• 한국축산식품학회지
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• 제34권1호
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• pp.65-72
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• 2014

This study is executed to develop probiotics which produce S-adenosyl-L-methionine (SAM), a methyl group donor of the 5-methyltetrahydrofolate methylation reaction within the animal cell. SAM is an essential substance for the synthesis, activation, and metabolism of hormones, neurotransmitters, nucleic acids, phospholipids, and cell membranes of animals. The SAM is also known as a nutritional supplement to improve brain functions of the human. In this study, the SAM-producing strains are identified in 18 types of salted fish, and then, the strains with excellent SAM productions are being identified, with 1 strain in the Enterococcus genus and 9 strains in the Bacillus genus. Strains with a large amount of SAM production include the lactic acid bacteria such as En. faecium and En. durans, En. sanguinicola, as well as various strains in the Bacillus genus. The SAM-overproducing strains show antibacterial activities with certain harmful microbes in addition to the weak acid resistances and strong bile resistances, indicating characteristics of probiotics. It is possible that the jeotgal-originated beneficial strains with overproducing SAM can be commercially utilized in order to manufacture SAM enriched foods.

• 한국미생물·생명공학회지
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• 제43권1호
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• pp.84-90
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• 2015

젓갈로부터 분리된 ampicillin 감수성, coagulase 음성 Staphylococcus 속 17 균주의 항생물질내성 및 위해성을 평가하였다. 15 균주는 한 종류 이상의 항생물질에 대한 내성을 나타내었고, penicillin G 내성 균주가 가장 높은 빈도로 검출되었다. PCR 증폭에 의한 항생물질내성 유전자 존재 확인 결과, trimethoprim 내성 관련 dfrA 유전자와 tetracycline 내성 관련 tetK 유전자를 각각 보유한 두 균주가 확인되었다. α 형 용혈활성은 검출되지 않았지만, 다섯 균주가 δ 형의 용혈을 나타내었고, 그 중 두 균주는 β 형 용혈활성을 나타냈으며, 12 균주가 biofilm을 형성하였다. 본 실험에서 수행한 모든 안전성 평가 결과는 균주 특이적으로 나타났다.

• Food Science and Biotechnology
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• 제17권4호
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• pp.857-860
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• 2008

S-Adenosyl-L-methionine (SAM) is a bioactive material used in the treatment of depression, osteoarthritis, and liver disease. To obtain lactic acid bacteria (LAB) producing high concentrations of SAM, LAB were isolated from commercial kimchi and from prepared kimchi products that contained shrimp jeotgal (fermented salty seafood) or sand lance jeotgal or that were fermented at 5 or $10^{\circ}C$, respectively, when pH was 4.2 to 4.8 and titratable acidity 0.6 to 0.9. Among the 179 LAB strains isolated from the fermented kimchi products, the genus Leuconostoc produced the highest intracellular level of SAM (1.58 mM) and Lactobacillus produced the second highest level (up to 1.47 mM) in the strain culture. This is the first study to quantify SAM in LAB isolated from fermented kimchi prepared by a general kimchi recipe. Ultimately, the selected strains (Leuconostoc mesentroides subsp. mesenteroides/dextranicum KSK417, L. mesentroides subsp. mesenteroides/dextranicum KJM401, and Lactobacillus bifermentans QMW327) could be useful as starters to manufacture fermented foods containing high levels of SAM.

• 한국미생물·생명공학회지
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• 제49권1호
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• pp.9-17
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• 2021

A γ-aminobutyric acid (GABA)-producing microorganism was isolated from galchi (hairtail fish, Trichiurus lepturus) jeotgal, a Korean salted and fermented seafood. The G144 isolate produced GABA excessively when incubated in MRS broth containing monosodium glutamate (MSG, 3%, w/v). G144 was identified as Lactobacillus brevis through 16S rRNA and recA gene sequencing. gadB and gadC encoding glutamate decarboxylase (GAD) and glutamate/GABA antiporter, respectively, were cloned and gadB was located downstream of gadC. The operon structure of gadCB was confirmed by reverse transcription (RT)-polymerase chain reaction. gadB was overexpressed in Escherichia coli and recombinant GAD was purified and its size was 54.4 kDa as evidenced by SDS-PAGE results. Maximum GAD activity was observed at pH 5.0 and 40℃ and the activity was dependent on pyridoxal 5'-phophate. The K_m and V_max of GAD were 8.6 mM and 0.01 mM/min, respectively.

• 한국수산과학회지
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• 제30권2호
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• pp.271-276
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• 1997

어획한 명태를 선상에서 가공할 때 부산물로 생산되는 선상창란을 대상으로 선도에 따른 제품의 관능검사와 제조과정 중의 수율을 조사함으로서 산업적으로 활용할 수 있는 원료창란의 VBN판정기준을 설정하고 아울러 초기 VBN 값이 각기 다른 원료창란으로부터 젓갈숙성중의 VBN, pH, 그리고 생균수 변화를 조사한 결과를 요약하면 다음과 같다. 1. 선상에서 처리한 원료창란은 창란젓갈 원료가 되는 위와 창자의 비율이 $72\%$로서 시료명태 내장중 위와 창자의 비율인 $18.5\%$보다 약 4배정도 높았다. 2. 동일한 시료명태에서 백색육과 내장의 VBN을 조사한 결과, 내장의 VBN이 백색육보다 최소 2배에서 최대 14.2배였고, 명태육과 내장의 선도 사이에는 일정한 상관 관계가 없었다. 3. 원료창란의 VBN이 $70{\pm}2mg\%$ 이하인 경우 위와 창자만을 정선한 수율은 $52\%$ 이상이었고, 제품의 관능 검사 결과도 양호하였다. 그러나$ VBN\;80{\pm}2mg\%$ 이상인 경우 정선창란의 수율은 $43\%$ 이하였으며 제품의 관능검사 결과도 불량하여 유통상품으로서 가치가 적었다. 4. 원료창란의 $VNB\;35{\pm}2mg\%$와 $70{\pm}2mg\%$인 시료를 60일 동안 숙성시키면서 VBN, pH 및 생균수 변화를 조사한 결과, VBN은 $50.06{\pm}2mg\%,\;87.71{\pm}2mg\%로서 두시료 모두 숙성초기보다 각각 $15.06mg{\%},\;17.51mg{\%}$ 증가하였으며, pH는 숙성초기에 7.12와 7.44이던 것이 각각 6.22, 6.15로 감소하였고, 생균수는 초기균수 $3.4{\times}10^3\;CFU/g$ 및 $3.4{\times}10^4\;CFU/g$에서 각각 $2.8{\times}10^4\;CFU/g$와$9.8{\times}10^5\;CFU/g$로 $1\~1.5-log-unit$가 증가하였다.