• Asian-Australasian Journal of Animal Sciences
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• 제17권10호
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• pp.1472-1478
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• 2004

The effects of microorganisms inoculated in beef, pork and chicken on the production of various biogenic amines (BA) were examined. Acinetobacter haemolyticus, Aeromonas hydrophila subsp. hydrophila, Alcaligenes faecalis subsp. faecalis, Bacillus cereus, Bacillus subtilis, Enterobacter aerogenes, Enterobacter cloacae, Escherichia coli, Lactobacillus alimentarius, Lactobacillus curvatus, Leuconostoc mesenteroides subsp. Mesenteroides, Proteus mirabilis, Proteus vulgaris, Pseudomonas aerugina, Salmonella enteritidis and Salmonella typhimurium were inoculated into beef, pork and chicken and incubated for 24 h at optimum temperatures of each bacterium. In ground beef, total amount of amines (TAA) produced was highest in the sample inoculated with Bacillus cereus, followed by Enterobacter cloacae. In ground pork, TAA was highest in the sample inoculated with Alcaligenes faecalis, followed by Enterobacter cloacae, Proteus vulgaris and Bacillus cereus. TAA of chicken breast was highest in the sample inoculated with Alcaligenes faecalis, followed by Bacillus cereus and Lactobacillus alimentarius while in chicken leg was the sample inoculated with Proteus vulgaris, followed by Enterobacter aerogenes, Enterobacter cloacae and Alcaligenes faecalis. Among biogenic amines produced, cadaverine (CAD) was detected at the highest level, followed by putrescine (PUT) and tyramine (TYM), their order being reversed by the kind of microorganism in beef and pork. In chicken breast and leg, CAD level was still the highest but PUT, TYM or PHM was the second highest, depending upon the kind of microorganism inoculated. In total, Alcaligenes faecalis, Enterobacter cloacae and Bacillus cereus were ones that produced a larger amount of BAs regardless of meat sources from different species.

• 한국축산식품학회지
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• 제38권1호
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• pp.152-161
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• 2018

This study aims to evaluate biogenic amine levels of bez sucuks (BS) produced with different meat:fat ratios. For this, three BS groups were manufactured with meat:fat ratios of 90:10 (BS10), 80:20 (BS20), and 70:30 (BS30). The pH and water activity values and biogenic amine amounts of sucuk samples were determined during processing and storage periods and the pH values of the initial mixtures of BS samples were in the range 5.51-5.74, decreasing to 4.72-4.94 by the $14^{th}$ day. The water activity values of BS samples showed significant decreases as a result of the drying stage and reached to range 0.913-0.935 on the $14^{th}$ day of processing (p<0.05). Although BS10 had the highest tyramine (434.12 mg/kg), histamine (5.69 mg/kg), cadaverine (12.48 mg/kg), putrescine (17.83 mg/kg), 2-phenylethylamine (15.43 mg/kg), and tryptamine (122.41 mg/kg) levels at the end of processing stage (p<0.05), spermine and spermidine levels did not differ between the BS samples due to their utilization of different meat:fat ratios (p>0.05). Similarly, the tryptamine (205.11 mg/kg), putrescine (43.57 mg/kg), and tyramine (766.23 mg/kg) levels of BS10 were higher than BS20 and BS30 samples at the end of storage (p<0.05). The results showed that BS10 with the highest meat ratio had the highest tryptamine, putrescine, and tyramine levels at the end of the processing and storage period.

• Toxicological Research
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• 제31권3호
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• pp.299-305
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• 2015

An HPLC analytical method was validated for the quantitative determination of biogenic amines in agricultural products. Four agricultural foods, including apple juice, Juk, corn oil and peanut butter, were selected as food matrices based on their water and fat contents (i.e., non-fatty liquid, non-fatty solid, fatty liquid and fatty solid, respectively). The precision, accuracy, recovery, limit of detection (LOD) and quantification (LOQ) were determined to test the validity of an HPLC procedure for the determination of biogenic amines, including tryptamine, ${\beta}$-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine, in each matrix. The LODs and LOQs for the biogenic amines were within the range of 0.01~0.10 mg/kg and 0.02~0.31 mg/kg, respectively. The relative standard deviation (RSD) of intraday for biogenic amine concentrations ranged from 1.86 to 5.95%, whereas the RSD of interday ranged from 2.08 to 5.96%. Of the matrices spiked with biogenic amines, corn oil with tyramine and Juk with putrescine exhibited the least accuracy of 84.85% and recovery rate of 89.63%, respectively, at the lowest concentration (10 mg/kg). Therefore, the validation results fulfilled AOAC criteria and recommendations. Subsequently, the method was applied to the analysis of biogenic amines in fermented agricultural products for a total dietary survey in Korea. Although the results revealed that Korean traditional soy sauce and Doenjang contained relatively high levels of histamine, the amounts are of no concern if these fermented agricultural products serve as condiments.

• Parasites, Hosts and Diseases
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• 제51권3호
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• pp.279-287
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• 2013

Autophagy is a process of cytoplasmic degradation of endogenous proteins and organelles. Although its primary role is protective, it can also contribute to cell death. Recently, autophagy was found to play a role in the activation of host defense against intracellular pathogens. The aims of our study was to investigate whether host cell autophagy influences Toxoplasma gondii proliferation and whether autophagy inhibitors modulate cell survival. HeLa cells were infected with T. gondii with and without rapamycin treatment to induce autophagy. Lactate dehydrogenase assays showed that cell death was extensive at 36-48 hr after infection in cells treated with T. gondii with or without rapamycin. The autophagic markers, LC3 II and Beclin 1, were strongly expressed at 18-24 hr after exposure as shown by Western blotting and RT-PCR. However, the subsequent T. gondii proliferation suppressed autophagy at 36 hr post-infection. Pre-treatment with the autophagy inhibitor, 3-methyladenine (3-MA), down-regulated LC3 II and Beclin 1. The latter was also down-regulated by calpeptin, a calpain inhibitor. Monodansyl cadaverine (MDC) staining detected numerous autophagic vacuoles (AVs) at 18 hr post-infection. Ultrastructural observations showed T. gondii proliferation in parasitophorous vacuoles (PVs) coinciding with a decline in the numbers of AVs by 18 hr. FACS analysis failed to confirm the presence of cell apoptosis after exposure to T. gondii and rapamycin. We concluded that T. gondii proliferation may inhibit host cell autophagy and has an impact on cell survival.

• 미생물학회지
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• 제55권3호
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• pp.258-267
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• 2019

본 연구의 목적은 탈지유와 두유 내에서 Enterobacter aerogenes CIH05의 증식과 바이오제닉 아민(biogenic amines, BA) 생성량에 대한 유산균의 영향을 조사하는 것이다. 갓김치로부터 분리된 Lactobacillus acidophilus GK20, Lactobacillus paracasei GK74 및 Lactobacillus planatrum GK81은 탈카르복시화 배지 상에서 BA를 생성하지 않았다. 탈지유와 두유 내에서 L. paracasei GK74에 의해 가장 많은 생균수와 항균 물질 생성량이 측정되었고, 반면 L. plantarum GK81은 다른 균주들에 비해 유의하게 낮은 균수와 항균 물질을 생산하였다. 유산, 과산화수소 및 박테리오신의 생산량은 Lactobacillus의 균종과 배지 종류에 의존하였다. 유산균과 유사하게 E. aerogenes CIH05도 단독 배양 시 두유보다 탈지유에서 더 많은 균수가 측정되었다. 탈지유와 두유 내에서 L. acidophilus GK20이나 L. paracasei GK74와 혼합 배양하거나 박테리오신 용액(300 AU/ml)을 처리했을 때 E. aerogenes CIH05의 균수가 유의하게 감소되었고 히스타민과 카다베린의 생성도 효과적으로 억제되었다.

• 한국수산과학회지
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• 제51권6호
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• pp.667-672
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• 2018

We investigated the quality of 12 commercial salted and fermented anchovy Engraulis japonicas sauces by measuring their chemical compositions, bacteria concentrations, and biogenic amine contents. The sauces had a moisture content of 67.13-69.83% (mean: 68.17%), salinity of 20.00-25.84% (mean: 22.29%), pH of 5.14-6.28 (mean: 5.63), volatile basic nitrogen content of 119.12-273.37 mg/100 g (mean: 199.19 mg/100 g), total nitrogen content of 0.82-1.50% (mean: 1.30%) and amino nitrogen content of 550.17-1,086.62 mg/100 g (mean: 774.99 mg/100 g). The viable cell counts ranged from not detected to $1.6{\times}10^3CFU/mL$, and the number of biogenic amine-forming bacteria was very low or undetectable. The tested samples contained 372.32-2,111,61 mg/kg (mean 813.48 mg/kg) histamine, 29.62-144.29 mg/kg (mean 98.14 mg/kg) cadaverine, 87.89-530.84 mg/kg (mean 329.91 mg/kg) tryptamine, 20.89-127.17 mg/kg (mean 60.49 mg/kg) putrescine, and 13.08-109.91 mg/kg (mean 57.74 mg/kg) tyramine. Whereas no spermidine or spermine was detected in any sample. These results strongly suggest the necessary of monitoring the biogenic amine contents of commercial salted and fermented anchovy sauces carefully to ensure consumer health.

• Journal of Microbiology and Biotechnology
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• 제31권7호
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• pp.949-955
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• 2021

Previously, our research group isolated Bifidobacterium breve IDCC4401 from infant feces as a potential probiotic. For this study, we evaluated the safety of B. breve IDCC4401 using genomic and phenotypic analyses. Whole genome sequencing was performed to identify genomic characteristics and investigate the potential presence of genes encoding virulence, antibiotic resistance, and mobile genetic elements. Phenotypic analyses including antibiotic susceptibility, enzyme activity, production of biogenic amines (BAs), and proportion of D-/L-lactate were evaluated using E-test, API ZYM test, high-performance liquid chromatography (HPLC), and D-/L-lactic acid assay respectively. The genome of B. breve IDCC4401 consists of 2,426,499 bp with a GC content of 58.70% and 2,016 coding regions. Confirmation of the genome as B. breve was provided by its 98.93% similarity with B. breve DSM20213. Furthermore, B. breve IDCC4401 genes encoding virulence and antibiotic resistance were not identified. Although B. breve IDCC4401 showed antibiotic resistance against vancomycin, we confirmed that this was an intrinsic feature since the antibiotic resistance gene was not present. B. breve IDCC4401 showed leucine arylamidase, cystine arylamidase, α-galactosidase, β-galactosidase, and α-glucosidase activities, whereas it did not show production of harmful enzymes such as β-glucosidase and β-glucuronidase. In addition, B. breve IDCC4401 did not produce any tyramine, histamine, putrescine, cadaverine, or 2-phenethylamine, which are frequently detected BAs during fermentation. B. breve IDCC4401 produced 95.08% of L-lactate and 4.92% of D-lactate. Therefore, our findings demonstrate the safety of B. breve IDCC 4401 as a potential probiotic for use in the food industry.

• Journal of Applied Biological Chemistry
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• 제63권4호
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• pp.375-385
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• 2020

본 연구의 목적은 발효 기간 동안 멸치 액젓과 새우젓으로 담근 배추 김치로부터 분리 동정된 바이오제닉 아민(biogenic amines, BA) 생성 유산균 단독 스타터 혹은 BA 분해 유산균과의 혼합 스타터로 제조한 김치의 품질 특성을 조사하고자 하였다. 발효가 진행될수록 새우젓 보다 멸치 액젓을 첨가하여 제조한 김치에서 유산균수, 산도 및 BA 함량이 높게 나타났다. 유산균이 생산하는 BA의 종류 및 생성량은 균종 보다는 균주에 의존적이었다. 분리 균주 중에서 가장 많은 양의 카다베린, 히스타민, 푸트레신 및 티라민은 각각 Leuconostoc mesenteroides MBK32, Lactobacillus brevis MBK34, Lactobacillus curvatus MBK31 및 Enterococcus faecalis SBK31로부터 생산되었다. BA 생성능과 분해능이 있는 스타터는 김치 내 유산균의 증식 속도와 산 생성능에 중요한 역할을 하였다. BA 생성균 단독 스타터에 의해 증가된 김치의 BA 함량은 BA 분해균과의 혼합 스타터에 의해 효과적으로 감소되었다.

• 한국응용과학기술학회지
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• 제37권6호
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• pp.1535-1544
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• 2020

본 연구의 목적은 3년 이상 숙성된 재래식 된장으로부터 아민 산화 효소를 생산하는 프로바이오틱 바실러스균을 분리 동정하는 것이다. 시료로부터 분리된 바이오제닉 아민(biogenic amines, BA) 생성균은 Bacillus sp. TS09, Bacillus licheniformis TS17, Bacillus subtilis TS19, Bacillus cereus TS23, Bacillus sp. TS30, Bacillus megaterium TS31, B. subtilis TS44, Bacillus coagulans TS46 및 Bacillus amyloliquefaciens TS59 등으로 동정되었다. 한편 동일한 시료로부터 분리된 B. subtilis TS04와 TS50은 인공 위액 및 담즙액에 대한 저항성, 장내 상피세포에 대한 부착능 및 BA 생성균(Bacillus sp. TS30 및 B. subtilis TS44)에 대한 박테리오신 생산 등의 프로바이오틱 활성을 나타내었다. 게다가 B. subtilis TS04와 TS50 균주가 생산한 아민 산화 효소에 의하여 카다베린, 푸트레신 및 티라민의 생성량을 감소시킬 수 있었으므로 이들은 BA 중독 위험을 낮출 수 있는 프로바이오틱스 소재로서의 활용 가치가 높을 것으로 판단된다.

• Journal of Microbiology and Biotechnology
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• 제31권1호
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• pp.25-32
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• 2021

Inflammatory reactions activated by lipopolysaccharide (LPS) of gram-negative bacteria can lead to severe septic shock. With the recent emergence of multidrug-resistant gram-negative bacteria and a lack of efficient ways to treat resulting infections, there is a need to develop novel anti-endotoxin agents. Antimicrobial peptides have been noticed as potential therapeutic molecules for bacterial infection and as candidates for new antibiotic drugs. We previously designed the 9-meric antimicrobial peptide Pro9-3 and it showed high antimicrobial activity against gram-negative bacteria. Here, to further examine its potency as an anti-endotoxin agent, we examined the anti-endotoxin activities of Pro9-3 and elucidated its mechanism of action. We performed a dye-leakage experiment and BODIPY-TR cadaverine and limulus amebocyte lysate assays for Pro9-3 as well as its lysine-substituted analogue and their enantiomers. The results confirmed that Pro9-3 targets the bacterial membrane and the arginine residues play key roles in its antimicrobial activity. Pro9-3 showed excellent LPS-neutralizing activity and LPS-binding properties, which were superior to those of other peptides. Saturation transfer difference-nuclear magnetic resonance experiments to explore the interaction between LPS and Pro9-3 revealed that Trp3 and Tlr7 in Pro9-3 are critical for attracting Pro9-3 to the LPS in the gram-negative bacterial membrane. Moreover, the anti-septic effect of Pro9-3 in vivo was investigated using an LPS-induced endotoxemia mouse model, demonstrating its dual activities: antibacterial activity against gram-negative bacteria and immunosuppressive effect preventing LPS-induced endotoxemia. Collectively, these results confirmed the therapeutic potential of Pro9-3 against infection of gram-negative bacteria.