• Title/Summary/Keyword: 16S-rRNA

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Isolation and Characterization of Plant-Derived Lactic Acid Bacteria as Potential Probiotic (잠재적 생균제로서 식물 젖산균의 분리 및 특성)

  • Kim, Jeong-Do;Park, Sung-Bo;Lee, Na-Ri;Jeong, Jin-Ha;Lee, Hee-Seob;Hwang, Dae-Youn;Lee, Jong-Sup;Jeong, Seong-Yun;Son, Hong-Joo
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.308-312
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    • 2011
  • Plant lactic acid bacteria were isolated from plant-associated fermentative foods and crops, and their probiotic properties were investigated. Isolates K27 and O2 were isolated from Kimchi and onion, and identified as Lactobacillus plantarum on the basis of 16S rRNA gene analysis. The two strains were highly resistant to acid (an MRS broth at pH 2.5), where the survival rates of L. plantarum K27 and L. plantarum O2 were 90.2% and 97.3%, respectively. L. plantarum K27 and L. plantarum O2 also showed high bile resistance to 0.5% oxgall, with a more than 70% survival rate. They showed an inhibitory effect against pathogenic strains of Escherichia coli KCCM 40880 and Pseudomonas aeruginosa ATCC 10145. The antibacterial effect of the two strains was probably due to the presence of lactic acid. ACE inhibitory activities of the two strains ranged from 72.8% to 80.6% in MRS broth. Notably, the two strains showed high ACE inhibitory activity (89.2~98.2%) in MRS broth containing 10% skim milk. Antioxidant activity was tested by DPPH radical scavenging activity, with antioxidant activities of the strains being in the range of 56.8~61.5%. The results obtained in this study suggest that L. plantarum K27 and L. plantarum O2 may be potential probiotic starter cultures with applications with fermentative products.

Antibacterial Activity and Probiotic Potential of Lactobacillus plantarum HKN01: A New Insight into the Morphological Changes of Antibacterial Compound-Treated Escherichia coli by Electron Microscopy

  • Sharafi, Hakimeh;Maleki, Hadi;Ahmadian, Gholamreza;Zahiri, Hossein Shahbani;Sajedinejad, Neda;Houshmand, Behzad;Vali, Hojatollah;Noghabi, Kambiz Akbari
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.225-236
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    • 2013
  • Among several bacteria examined, an antibacterial-producing Lactobacillus strain with probiotic characteristics was selected and identified based on 16S rRNA gene sequencing. Subsequent purification and mode of action of the antibacterial compounds on target cells including E. coli were investigated. Maximum production of the antibacterial compound was recorded at 18 h incubation at $30^{\circ}C$. Interestingly, antibacterial activity remained unchanged after heating at $121^{\circ}C$ for 45 min, 24 h storage in temperature range of $70^{\circ}C$ to room temperature, and 15 min exposure to UV light, and it was stable in the pH of range 2-10. The active compounds were inactivated by proteolytic enzymes, indicating their proteinaceous nature, and, therefore, referred to as bacteriocin-like inhibitory substances. Isolation and partial purification of the effective agent was done by performing ammonium sulfate precipitation and gel filtration chromatography. The molecular mass of the GFC-purified active compound (~3 kDa) was determined by Tris-Tricine SDS-PAGE. To predict the mechanisms of action, transmission electron microscopy (TEM) analysis of ultrathin sections of E. coli before and after antibacterial treatment was carried out. TEM analysis of antibacterial compounds-treated E. coli demonstrated that the completely altered bacteria appear much darker compared with the less altered bacteria, suggesting a change in the cytoplasmic composition. There were also some membrane-bound convoluted structures visible within the completely altered bacteria, which could be attributed to the response of the E. coli to the treatment with the antibacterial compound. According to the in vivo experiments oral administration of L. plantarum HKN01 resulted in recovery of infected BALB/c mice with Salmonella enterica ser. Typhimurium.

Fermentation of Cucurbita maxima Extracts with Microganisms from Kimchi (김치 유래 유산균을 이용한 단호박 발효음료 제조 기술 개발)

  • Roh, Hyun-Ji;Kim, Gi-Eun
    • KSBB Journal
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    • v.24 no.2
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    • pp.149-155
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    • 2009
  • 19 strains, which could be identified as Lactobacillus sp. were isolated. The Cucurbita maxima has been known as a traditional healthy food and variable positive effects on the human body were already reported. In this study we tried to develop a production process for a healthy fermented drink with Cucurbita maxima and strains originated from Kimchi. Many kinds of lacctobacci species existed in the fermented food cannot survive in the acidic conditions in the stomach. So we tried to search and select a strain, which can arrive to the small intestine. A species of a Lactobacillus named as C332 was identifed as Lactobacillus plantarum and selected for the fermentation process. With the treatment with artificial gastric juice and artificial bile the survival rate of the cells could be calculated. The physiological characteristics at the variable conditions have been tested. After fermentation process the sensoric tests on the product with panels were tried. The most of the cells could survive in the acidic conditions and falcultive anaerobe. Especially some antibacterial effects aganinst E.coli were also found. With all kinds of the results from our research the fermented Cucurbita maxima drink can be a successful item in the market.

Isolation and Characterization of Comamonase sp. and Microbacterium sp. from Deep Blue Sediment Dye of Polygoum tinctoria, Niram (쪽 염료 니람으로부터 Comamonas sp.와 Microbacterium sp.의 분리 및 특성분석)

  • Jang, Seong Eun;Lee, Nam Keun;Lee, Yuri;Choi, Mee-Sung;Jeong, Yong-Seob
    • KSBB Journal
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    • v.28 no.1
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    • pp.60-64
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    • 2013
  • Two strains were isolated from the traditional Deep Blue Sediment Dye of Polygoum tinctoria, Niram, and temporarily named Niram A and Niram B, respectively. The phylogenetic analysis revealed that strain Niram A and B were closely related to the members of the genus Comamonas and Microbacterium, respectively. Strain Niram A exhibited the highest 16S rRNA gene sequence similarity to C. aquatica LMG $2370^T$ (98.06%). Strain Niram B showed 100% homology with M. oxydans DSM 20578T and M. maritypicum DSM $12512^T$. The growth of the strain Niram A and B was not inhibited in Niram medium containing high calcium concentration without free sugar as carbon source. The reducing Niram is greenish. Therefore, the reducing ability on the Niram of the strains Niram A and B were determined with the color difference of the $a^*$ values of Niram fermented-fluids. The $a^*$ value indicates the level of redness (positive value) or greenness (negative value). The green color is increasing towards the negative value. In all samples fermented for 10 days, the $a^*$ values among samples were no significant difference. However, samples fermented for 15 days have an appreciable change. After fermentation for 15 days, the control Niram sample had $-3.96{\pm}0.02$ of the $a^*$ value. On the other hand, the Niram samples fermented with the strain Niram A and B showed $-4.20{\pm}0.02$ of the $a^*$ value and $-7.86{\pm}0.03$ of the $a^*$ value, respectively. In the reducing ability on the Niram, the strain Niram B was significantly better than the strain Niram A.

Growth Suppression of Microcystis aeruginosa by Pseudomonas aeruginosa AJ1 (Pseudomonas aeruginosa AJ1에 의한 Microcystis aeruginosa의 성장제어)

  • Kim, Sun-Jung;Lee, Sang-Seob
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.362-367
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    • 2009
  • Among total 176 strains with antialgal effects isolated from So-ok stream in Korea, Pseudomonas aeruginosa AJ1 showed the highest removal efficiency for an algal species Microcystis aeruginosa (clear zone of diameter 50.0 mm on algal lawn after 20 days). The algal growth was suppressed even when the supernatant of AJ1 culture was applied, suggesting that extracellular substances are responsible for its antialgal activity. The removal activity of AJ1 was optimal under the following condition: pH 8, $30^{\circ}C$, and mannitol as a carbon source. The antialgal activity of AJ1 appeared to be dependent of the growth phase of M. aeruginosa, i.e., the highest at the early phase, but not its own phase. As expected, the algicidal effect was improved as the amount of the treated supernatant was increased; the highest removal efficiency (80.3%) was achieved when 40 ml/L of the supernatant was used. Interestingly, however, the removal rate was opposite. The highest removal rate ($8.2{\mu}g$ chl-a/ml supernatant/day) was achieved when low concentration (10 ml/L) was applied. These results suggest that P. aeruginosa AJ1 is a promising biological agent to control the problematic algal bloom.

Comparison of Conventional Culture Method and Real-time PCR for Detection of Yersinia enterocolitica in Sausage and Vegetable Salad (소시지와 야채 샐러드에서 Yersinia enterocolitica 검출을 위한 배지법과 real-time PCR법의 비교)

  • Kim, Yun-Gyeong;Chon, Jung-Whan;Lee, Jae-Hoon;Kwak, Hyo-Sun;Hwang, In-Gyun;Seo, Kun-Ho
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.133-136
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    • 2013
  • The purpose of this study was to compare a conventional culture method and real-time PCR for the detection of Yersinia enterocolitica (Y. enterocolitica) in sausage and in vegetable salad. Food samples inoculated with Y. enterocolitica were enriched in peptone-sorbitol bile-broth, and swabs were then streaked onto cefsulodin-irgasan-novobiocin agar. Biochemical tests for suspected colonies were performed with an API 20E strip. In parallel, real-time PCR was performed, targeting the 16S rRNA gene using 1 mL of enrichment broth. In sausage, the number of positive samples detected by culture method (49 out of 60) was similar (p>0.05) with that of real-time PCR (50 out of 60). However, the number of positive samples of real-time PCR (26 out of 60) was significantly higher (p<0.05) than that of the conventional culture method (6 out of 60) in vegetable salad. Real-time PCR could be an effective screening tool for detecting Y. enterocolitica, particularly in food samples with high levels of background flora, such as a vegetable salad.

Optimal Culture Conditions and Food Waste Decomposition Effects of Mixed Strains Separated from Traditional Fermented Food and Soils (전통발효식품과 토양으로부터 분리된 혼합균주의 최적생육조건 및 음식물쓰레기 분해 효과)

  • Kim, Min-Sun;Kim, Hee-Jeong;Jung, Eun-Seon;Park, Ju-Yong;Chae, Jong-Chan;Hwang, Kwontack;Lee, Seung-Je
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.285-292
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    • 2018
  • In this study, for the purpose of decomposing food waste, the strain was screened from traditional fermented food and soils. The enzyme activity (protease, amylase, cellulase, lipase) experiment was carried out using the paper disc method in 212 strains isolated from 5% NaCl media. Among them, only the strains having enzyme activity of more than 2 (soil) or more than 4 (traditional fermented food) with the halozone of enzyme activity of 15 mm or more were selected first, and microorganism identification through 16S rRNA sequencing was performed. Finally, were identified such as Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus siamensis, Bacillus licheniformis, Bacillus aquimaris, Bacillus megaterium, Bacillus koreensis, Bacillus stratoshericus, Bacillus aryabhattai, Bacillus safensis, Marinobacter hydrocarbonoclasticus. 11 species of mixed strains were confirmed that the culture time was 24 hours, the incubation temperature was $30^{\circ}C$ and the optimum pH was 7.0. In order to confirm the degree of decomposition of standard food wastes (100 g) by treating 11 kinds of mixed strains (25%), solid content of more than $2000{\mu}m$ was determined to be 103 g for the sterilized water group and 18 g for the mixed strains group. And the rest was decomposed to a size of less than $2000{\mu}m$.

Microbial Diversity and Community Analysis in Lettuce or Cucumber Cultivated Greenhouse Soil in Korea (상추 및 오이 시설재배 토양의 미생물 다양성 분석)

  • Kim, Byung-Yong;Weon, Hang-Yeon;Park, In-Cheol;Lee, Sang-Yeob;Kim, Wan-Gyu;Song, Jae-Kyeong
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.6
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    • pp.1169-1175
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    • 2011
  • The soil chemical properties, microbial community structures and biochemical properties of lettuce or cucumber-cultivated greenhouse soil samples were analyzed to assess soil health and characterize microbial distribution in 8 locations in Korea. Although most of chemical properties were within the soil management guidelines, the available phosphate, and the contents of exchangeable potassium and calcium were higher than those of recommended levels. In the culture-dependent analysis, 841 bacterial strains were isolated from the greenhouse soils and were identified at the genus level by 16S rRNA gene sequences analysis. The dominant bacterial genera were Bacillus (35.7%), Microbacterium (9.3%), Arthrobacter (5.7%) and Lysobacter (5.1%). The abundance of pseudomonads was highly variable depending on the soil samples. In the culture-independent analysis, soil microbial community was investigated by using phospholipid fatty acid (PLFA) method. Principal component analysis (PCA) showed that a specific grouping for microbial community structure in the greenhouse soils was not observed based on cultivated crops and investigated sites. The results revealed that the greenhouses soils examined are relatively sound managed in terms of soil chemical contents and microbial properties.

Enhancing Butyrate Production, Ruminal Fermentation and Microbial Population through Supplementation with Clostridium saccharobutylicum

  • Miguel, Michelle A.;Lee, Sung Sill;Mamuad, Lovelia L.;Choi, Yeon Jae;Jeong, Chang Dae;Son, Arang;Cho, Kwang Keun;Kim, Eun Tae;Kim, Sang Bum;Lee, Sang Suk
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1083-1095
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    • 2019
  • Butyrate is known to play a significant role in energy metabolism and regulating genomic activities that influence rumen nutrition utilization and function. Thus, this study investigated the effects of an isolated butyrate-producing bacteria, Clostridium saccharobutylicum, in rumen butyrate production, fermentation parameters and microbial population in Holstein-Friesian cow. An isolated butyrate-producing bacterium from the ruminal fluid of a Holstein-Friesian cow was identified and characterized as Clostridium saccharobutylicum RNAL841125 using 16S rRNA gene sequencing and phylogenetic analyses. The bacterium was evaluated on its effects as supplement on in vitro rumen fermentation and microbial population. Supplementation with $10^6CFU/ml$ Clostridium saccharobutylicum increased (p < 0.05) microbial crude protein, butyrate and total volatile fatty acids concentration but had no significant effect on $NH_3-N$ at 24 h incubation. Butyrate and total VFA concentrations were higher (p < 0.05) in supplementation with $10^6CFU/ml$ Clostridium saccharobutylicum compared with control, with no differences observed for total gas production, $NH_3-N$ and propionate concentration. However, as the inclusion rate (CFU/ml) of C. saccharobutylicum was increased, reduction of rumen fermentation values was observed. Furthermore, butyrate-producing bacteria and Fibrobacter succinogenes population in the rumen increased in response with supplementation of C. saccharobutylicum, while no differences in the population in total bacteria, protozoa and fungi were observed among treatments. Overall, our study suggests that supplementation with $10^6CFU/ml$ C. saccharobutylicum has the potential to improve ruminal fermentation through increased concentrations of butyrate and total volatile fatty acid, and enhanced population of butyrate-producing bacteria and cellulolytic bacteria F. succinogenes.

Effect of commercially purified deoxynivalenol and zearalenone mycotoxins on microbial diversity of pig cecum contents

  • Reddy, Kondreddy Eswar;Kim, Minji;Kim, Ki Hyun;Ji, Sang Yun;Baek, Youlchang;Chun, Ju Lan;Jung, Hyun Jung;Choe, Changyong;Lee, Hyun Jeong;Kim, Minseok;Lee, Sung Dae
    • Animal Bioscience
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    • v.34 no.2
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    • pp.243-255
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    • 2021
  • Objective: Deoxynivalenol (DON) and zearalenone (ZEN) are mycotoxins that frequently contaminate maize and grain cereals, imposing risks to the health of both humans and animals and leading to economic losses. The gut microbiome has been shown to help combat the effects of such toxins, with certain microorganisms reported to contribute significantly to the detoxification process. Methods: We examined the cecum contents of three different dietary groups of pigs (control, as well as diets contaminated with 8 mg DON/kg feed or 0.8 mg ZEN/kg feed). Bacterial 16S rRNA gene amplicons were acquired from the cecum contents and evaluated by next-generation sequencing. Results: A total of 2,539,288 sequences were generated with ~500 nucleotide read lengths. Firmicutes, Bacteroidetes, and Proteobacteria were the dominant phyla, occupying more than 96% of all three groups. Lactobacillus, Bacteroides, Megasphaera, and Campylobacter showed potential as biomarkers for each group. Particularly, Lactobacillus and Bacteroides were more abundant in the DON and ZEN groups than in the control. Additionally, 52,414 operational taxonomic units were detected in the three groups; those of Bacteroides, Lactobacillus, Campylobacter, and Prevotella were most dominant and significantly varied between groups. Hence, contamination of feed by DON and ZEN affected the cecum microbiota, while Lactobacillus and Bacteroides were highly abundant and positively influenced the host physiology. Conclusion: Lactobacillus and Bacteroides play key roles in the process of detoxification and improving the immune response. We, therefore, believe that these results may be useful for determining whether disturbances in the intestinal microflora, such as the toxic effects of DON and ZEN, can be treated by modulating the intestinal bacterial flora.