• Title/Summary/Keyword: 16S-rRNA

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Characterization of a Glutamate Decarboxylase (GAD) from Enterococcus avium M5 Isolated from Jeotgal, a Korean Fermented Seafood

  • Lee, Kang Wook;Shim, Jae Min;Yao, Zhuang;Kim, Jeong A;Kim, Hyun-Jin;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1216-1222
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    • 2017
  • To develop starters for the production of functional foods or materials, lactic acid bacteria producing ${\gamma}-aminobutyric$ acid (GABA) were screened from jeotgals, Korean fermented seafoods. One isolate producing a high amount of GABA from monosodium $\text\tiny{L}$-glutamate (MSG) was identified as Enterococcus avium by 16S rRNA gene sequencing. E. avium M5 produced $18.47{\pm}1.26mg/ml$ GABA when incubated for 48 h at $37^{\circ}C$ in MRS broth with MSG (3% (w/v)). A gadB gene encoding glutamate decarboxylase (GAD) was cloned and overexpressed in E. coli BL21 (DE3) using the pET26b (+) expression vector. Recombinant GAD was purified through a Ni-NTA column and the size was estimated to be 53 kDa by SDS-PAGE. Maximum GAD activity was observed at pH 4.5 and $55^{\circ}C$and the activity was dependent on pyridoxal 5'-phosphate. The $K_m$ and $V_{max}$ values of GAD were $3.26{\pm}0.21mM$ and $0.0120{\pm}0.0001mM/min$, respectively, when MSG was used as a substrate. Enterococcus avium M5 secretes a lot of GABA when grown on MRS with MSG, and the strain is useful for the production of fermented foods containing a high amount of GABA.

Insights into the Gut Microbiota of Freshwater Shrimp and Its Associations with the Surrounding Microbiota and Environmental Factors

  • Zhao, Yanting;Duan, Cuilan;Zhang, Xu-xiang;Chen, Huangen;Ren, Hongqiang;Yin, Ying;Ye, Lin
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.946-956
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    • 2018
  • The gut microbiota of aquatic animals plays a crucial role in host health through nutrient acquisition and outcompetition of pathogens. In this study, on the basis of the high-throughput sequencing of 16S rRNA gene amplicons, we examined the bacterial communities in the gut of freshwater shrimp (Macrobrachium nipponense) and in their living environments (sediment and pond water) and analyzed the effects of abiotic and biotic factors on the shrimp gut bacterial communities. High bacterial heterogeneity was observed in the freshwater shrimp gut samples, and the result indicated that both the surrounding bacterial community and water quality factors (particularly dissolved oxygen and temperature) could affect the shrimp gut bacterial community. Despite the observed heterogeneity, 57 genera, constituting 38-99% of the total genera in each of the 40 shrimp gut samples, were identified as the main bacterial population in the gut of M. nipponense. In addition, a high diversity and abundance of lactic acid bacteria (26 genera), which could play significant roles in the digestion process in shrimp, were observed in the shrimp gut samples. Overall, this study provides insights into the gut bacterial communities of freshwater shrimp and basic information for shrimp farming regarding the application of probiotics and disease prevention.

Zerumbone Restores Gut Microbiota Composition in ETBF Colonized AOM/DSS Mice

  • Cho, Hye-Won;Rhee, Ki-Jong;Eom, Yong-Bin
    • Journal of Microbiology and Biotechnology
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    • v.30 no.11
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    • pp.1640-1650
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    • 2020
  • Colorectal cancer (CRC) is the leading cause of common malignant neoplasm worldwide. Many studies have analyzed compositions of gut microbiota associated with various diseases such as inflammatory bowel diseases (IBD) and colon cancer. One of the most representative bacteria involved in CRC is enterotoxigenic Bacteroides fragilis (ETBF), a species belonging to phylum Bacteroidetes. We used ETBF colonized mice with azoxymethane (AOM)/dextran sulphate sodium (DSS) and zerumbone, a compound with anti-bacterial effect, to determine whether zerumbone could restore intestinal microbiota composition. Four experimental groups of mice were used: sham, ETBF colonized AOM/DSS group, ETBF colonized AOM/DSS group zerumbone 60 mg kg-1 (ETBF/AOM/DSS + Z (60)), and only zerumbone (60 mg kg-1)-treated group. We performed reversible dye terminators-based analysis of 16S rRNA gene region V3-V4 for group comparison. Microbiota compositions of ETBF/AOM/DSS + Z (60) group and ETBF colonized AOM/DSS group not given zerumbone were significantly different. There were more Bacteroides in ETBF/AOM/DSS + Z (60) group than those in ETBF colonized AOM/DSS group, suggesting that B. fragilis could be a normal flora activated by zerumbone. In addition, based on linear discriminant analysis of effect size (LEfSe) analysis, microbial diversity decreased significantly in the ETBF colonized AOM/DSS group. However, after given zerumbone, the taxonomic relative abundance was increased. These findings suggest that zerumbone not only influenced the microbial diversity and richness, but also could be helpful for enhancing the balance of gut microbial composition. In this work, we demonstrate that zerumbone could restore the composition of intestinal microbiota.

Probiotic Properties of Pediococcus pentosaceus SH-10 Isolated from the Hard Clam Meretrix meretrix Shikhae (백합(Meretrix meretrix)식해에서 분리한 Pediococcus pentosaceus SH-10의 생균제적 특성)

  • Song, Hyun-Jung;Kim, Kang-Jin;Kim, Hee-Dai;Yoo, Jung-Hee;Koo, Jae-Geun;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.6
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    • pp.605-611
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    • 2011
  • This study examined the suitability of characteristics of potential strains of probiotic bacteria. Among 25 lactic acid bacteria isolated from Korean traditional fermented food, the Hard Clam Meretrix meretrix Shikhae, the SH-10 strain, which exhibited superior resistance to low pH and bile salts, was selected as a potential probiotic bacteria. By examining carbohydrate utilization, morphological properties, and the 16S rRNA gene sequence, the SH-10 strain was identified as Pediococcus pentosaceus (hereafter, P. pentosaceus SH-10). P. pentosaceus SH-10 was resistant to amikacin, cefotetan, ciprofloxacin, gentamicin, kanamycin, nalidixic acid, streptomycin, and vancomycin. Tests of antimicrobial activities against pathogens such as Bacillus cereus, Listeria monocytogenes, Salmonella choleraesuis, and Staphylococcus aureus, indicated that P. pentosaceus SH-10 inhibited the growth of pathogenic bacteria. These results suggest that P. pentosaceus SH-10 can be developed as a probiotic bacteria.

Diagnosis of Bovine Theileriosis by Direct PCR and Electrophoresis from Whole Blood Without DNA Extraction (DNA 추출없이 전혈을 이용한 PCR-전기영동법에 의한소의 타일레리아병 진단)

  • Seong Ho Kang;Sangmin Jang;Joon-Seok Chae;Yongseong Kim
    • Journal of the Korean Chemical Society
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    • v.47 no.2
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    • pp.127-132
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    • 2003
  • We have developed a direct polymerase chain reaction (PCR) and electrophoresis method for the diagnosis of bovine theileriosis from whole blood DNA analysis without DNA extraction. The technique empolyed a FoLT (formamide, low temp.) technique and was utilized in the diagnosis of bovine theileriosis. Formamide solubilize the blood cells, and the lowered incubation temperatures reduced protein coagulation. 100-200 nL of whole blood and PCR reagents were introduced directly into a PCR tube. After the amplification, the PCR product (816-bp DNA) was introduced into the electrophoresis system. The results of this analysis were consist with those obtained using purified DNA.

High prevalence of Enterococcus spp. from dogs with otitis externa

  • Jo, Hyun-Jung;Chae, Hee-Sun;Kim, Hyun-Ju;Kim, Min-Ju;Park, Gyu-Nam;Kim, Sang-Hun;Chang, Kyung-Soo
    • Korean Journal of Veterinary Service
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    • v.35 no.2
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    • pp.99-104
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    • 2012
  • Otitis externa (OE) is a frequent disease in the ear canals of dogs. To identify the pathogens causing OE in dogs and to determine their antimicrobial resistances, specimens were collected from animal hospitals in Daejeon. The isolates were examined by morphological and biochemical tests, 16S rRNA analysis and antimicrobial susceptibility tests. We analyzed correlation between the isolated pathogens and external factors of dogs such as breed, age, gender, ear mite, hair in ears and experience with antibiotic therapy. Thirty three strains of bacteria were isolated from 26 of the 68 heads of dogs with OE. The most isolated bacteria were Enterococcus faecalis (E. faecalis) followed by Staphylococcus aureus (Sta. aureus), Sta. pseudointermedius, E. faecium, E. avium and Streptococcus canis (Strep. canis) in order of frequency of occurrence. Isolation frequency of Enterococcus spp. and Staphylococcus spp. were 51.5% and 45.5%, respectively. E. faecalis and E. faecium isolates showed VanB phenotype, which is resistant to vancomycin but sensitive to teicoplanin were 58% and 25%, respectively. Nine isolates among total twelve isolates of E. faecalis were isolated from the dogs treated with antibiotics. There was no methicillin-resistant Sta. aureus (MRSA), but were MR-Sta. pseudointermedius (MRSP) (57.1%) and vancomycin-resistant (VR)-Sta. pseudointermedius (14.3%) (VRSP) showing VanB phenotype. However, vanA, vanB and vanC genes were not detected in VR isolates from the dogs. Taken together, VR-Enterococcus spp. (VRE) is one of the major pathogens in domestic animals, as well as community-and hospital-acquired infection.

Functional Characterization of Lactobacillus sakei JK-17 Isolated from Long-term Fermented Kimchi, Muk Eun Ji (장기간 발효 김치인 묵은지에서 분리한 Lactobacillus sakei JK-17의 기능성 조사)

  • Kim, Dong-Seon;Cho, Hyeong-Woo;Kim, Dae-Han;Oh, Kye-Heon
    • KSBB Journal
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    • v.28 no.1
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    • pp.18-23
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    • 2013
  • The purpose of this work was to investigate the several functional characteristics of Lactobacillus sakei JK-17 isolated from long-term fermented kimchi, Muk Eun Ji. Initially, phylogenetic analysis using 16S rRNA sequencing was performed to identify the isolate JK-17, and the strain could be assigned to Lactobacillus sakei and designated as L. sakei JK-17. The strain was registered in GenBank as [JX841311]. The changes of bacterial growth and residual organic acids were monitored and HPLC was used to measure quantitatively two organic acids, lactic acid and acetic acid, produced in the culture during 84 hours of incubation. During the incubation period, several functional characteristics of L. sakei JK-17 were examined. L. sakei JK-17 culture depleted nitrite concentration 94.75%. Antioxidant activity of cultural supernatants of L. sakei JK-17 was approx. 53.8%, and ${\beta}$-galactosidase activities were 0.243 units/mL at pH 7.0 and 0.387 units/mL at pH 4.1, respectively. The antibacterial activities against food-poisoning causing bacteria were examined with 20-fold concentrated culture supernatants from L. sakei JK-17 and the antibacterial effects were clearly observed against all bacteria tested in this work.

Occurrence of Sweet-potato Whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae) and Its Response to Insecticide in Gyeonggi Area (경기지역에서 담배가루이의 발생 및 약제반응)

  • Lee, Young-Su;Kim, Jin-Young;Hong, Soon-Sung;Park, Jungan;Park, Hong-Hyun
    • Korean journal of applied entomology
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    • v.51 no.4
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    • pp.377-382
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    • 2012
  • This study investigated the occurrence of sweet potato whitefly, Bemisia tabaci affecting cucumber, eggplant and red pepper, as well as sweet potato species, and its response to insecticides in Gyeonggi province from 2010 to 2011. Sweet potato whitefly is widespread throughout the southern part of Gyeonggi province. Most regional populations of B. tabaci belong to biotype Q having been reported in the south Korea since 2005, but in Goyang mixed populations of two biotypes (B and Q) were found. Survey results of Tomato Yellow Leaf Curl Virus (TYLCV) disease that was vectored by B. tabaci indicated that this virus disease was not spread throughout the Gyeonggi province. Biotype Q of B. tabaci was found to be resistant to neonicotinoid insecticides, whereas biotype B was highly susceptible to them.

MOLECULAR IDENTIFICATION OF BACTERIA FROM OSTEOMYELITIS OF THE JAWS (분자생물학적 기법을 이용한 악골 골수염 병소의 세균 동정)

  • Kim, Mi-Sung;Kim, Su-Gwan;Chung, Hae-Man;Kim, Sang-Gon;Kook, Joong-Ki;Kim, Mi-Kwang;Kim, Hwa-Sook;Yoo, So-Young
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.29 no.1
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    • pp.48-55
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    • 2003
  • The purpose of this study was to isolate and identify the bacteria in osteomyelitis lesion of 3 patients. Two lesions were due to the post-infection after extraction. The other was resulted from mal-fixation of both sides of mandibular angles. Pus samples were collected by needle aspiration from the lesion and examined by culture method. Bacterial culture was performed in three culture systems (anaerobic, $CO_2$, and aerobic incubator). Identification of the bacteria was performed by 16S rRNA gene cloning and nucleotide sequencing method. Our results showed that Streptococci species was predominantly isolated in both lesions of extraction socket. Only one species (Proteus vulagris) was detected in lesion of mandibular angle. This study was not sufficient to identify the causative bacteria in those osteomyelitis. However, our data may be offered the clue to solve the problem.

Antagonistic Effects of the Bacterium Alcaligenes sp. HC12 on Browning Disease Caused by Pseudomonas agarici (버섯 세균성회색무늬병균(Pseudomonas agarici)에 대한 Alcaligenes sp. HC12의 항균활성)

  • Lee, Chan-Jung;Moon, Ji-Won;Cheong, Jong-Chun;Kong, Won-Sik
    • The Korean Journal of Mycology
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    • v.44 no.3
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    • pp.171-175
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    • 2016
  • A gram-negative bacterium was isolated from spent substrates of Agaricus bisporus and showed significant antagonistic activity against Pseudomonas agarici. The bacterium was identified as Alcaligenes sp. based on cultural, biochemical, physiological characteristics and a 16S rRNA sequence analysis. The isolate is saprophytic, but not parasitic or pathogenic on cultivated mushroom, whereas it showed strong inhibitory effects against P. agarici cells in vitro. The control efficacy of Alcaligenes sp. HC12 against brown blotch of P. agarici was up to 63% on Agaricus bisporus. The suppressive bacterium may be useful for the development of biocontrol systems.