• Title/Summary/Keyword: 16S rRNA Gene

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Isolation and Characterization of Kimchi Lactic Acid Bacteria Showing Anti-Helicobacter pylori Activity (Helicobacter pylori 억제능 김치 유산균의 분리와 특성 규명)

  • Lee, Youl;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.36 no.2
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    • pp.106-114
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    • 2008
  • One bacterium, which showed strong antagonistic activity against H. pylori KCCM 41756, was isolated from kimchi. The strain NO1 was designated as Lactobacillus plantarum NO1 based on Gram staining, biochemical properties, and 16S rRNA gene sequencing. The culture medium $(2{\sim}4{\mu}g/ml)$ of Lb. plantarum NO1 reduced $(40{\sim}60%)$ the urease activity of H. pylori KCCM 41756. Lb. plantarum NO1 inhibited the binding of H. pylori to human gastric cancer cell line, AGS cells, by more than 33%. Lb. plantarum NO1 exhibited high viability (maintained initial viable cell count of $10^9CFU/ml$) in 0.05 M sodium phosphate buffer (pH 3.0) for 2 h, in artificial gastricjuice for 2 h and in 0.3%, 0.5% oxgall for 24 h. Hemolysis phenomena did not observed when Lb. plantarum NO1 was incubated in the blood agar media. We concluded that Lb. plantarum NO1 can be a good candidate as a probiotic, harboring anti-H. pylori activity.

Bacterial core community in soybean rhizosphere (콩 근권의 핵심 세균 군집)

  • Lee, Youngmi;Ahn, Jae-Hyung;Choi, Yu-Mi;Weon, Hang-Yeon;Yoon, Jung-Hoon;Song, Jaekyeong
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.347-354
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    • 2015
  • Soybean is well known to be originated from Korea and far-east Asian countries, and studies of many root nodule bacteria associated with soybean have mainly-focused on nitrogen fixation, but much less study was carried out on bacterial community in the rhizosphere of soybean. In this study, we analyzed the bacterial community in rhizosphere of Korean soybean, Daepungkong using the pyrosequencing method based on the 16S rRNA gene to characterize the change of the rhizosphere community structure according to the growth stages of soybeans and to elucidate bacterial core community in rhizosphere of soybean. Our results revealed that bacterial community of rhizosphere soil differed from that of bulk soil and was composed of a total of 21 bacterial phyla. The predominant phylum in the rhizosphere of soybean was Proteobacteria (36.6-42.5%) and followed by Acidobacteria (8.6-9.4%), Bacteroidetes (6.1-10.9%), Actinobacteria (6.4-9.8%), and Firmicutes (5.7-6.3%). The bacterial core community in soybean rhizosphere was mainly composed of the operational taxonomic units (OTUs) belonging to the phylum Proteobacteria throughout all growth stages. The OTU00006 belonged to the genus Bradyrhizobium had the highest abundance and Steroidobacter, Streptomyces, Devosia were followed. These results show that bacterial core community in soybean rhizosphere was mainly composed of OTUs associated with plant growth promotion and nutrient cycles.

Functional evaluation of sourdough containing lactic acid bacteria isolated from sliced radish kimchi (깍두기로부터 분리된 유산균으로 제조한 사워도우의 기능성 평가)

  • Lim, Eun-Seo;Kim, Young-Mog;Lee, Eun-Woo
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.180-192
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    • 2017
  • The purpose of this study is to investigate the antioxidative and antimicrobial activities of sourdough fermented with the lactic acid bacteria (LAB) isolated from sliced radish kimchi. According to 16S rRNA gene sequence analysis, the isolated lactic acid bacteria were categorized as Leuconostoc dextranicum SRK03, Lactobacillus brevis SRK15, Pediococcus halophilus SRK22, Lactobacillus acidophilus SRK30, Lactobacillus plantarum SRK38, Leuconostoc citreum SRK 42, and Lactobacillus delbrueckii SRK60 with sequence similarity of 99%. After fermentation with L. dextranicum SRK03, L. acidophilus SRK30, L. plantarum SRK38 or L. delbrueckii SRK60 and Saccharomyces cerevisiae KCTC 7246 at $30^{\circ}C$ for 24 h LAB and yeast in sourdough were present at levels of $10^9$ and $10^7CFU/g$, respectively. In particular, the titratable acidity and ethanol and exopolysaccharide contents of sourdough fermented with L. dextranicum SRK03 were also significantly (P < 0.05) higher than those of sourdough fermented with L. acidophilus SRK30, L. plantarum SRK38, or L. delbrueckii SRK60. The sourdough fermented with L. dextranicum SRK03 and L. acidophilus SRK30 showed not only good DPPH radical-scavenging capacity but anti-lipid peroxidation activity. In addition, the viable counts of Bacillus cereus ATCC 11778 and Staphylococcus aureus ATCC 6538 in sourdough during storage for 5 days at $25^{\circ}C$ were significantly (P < 0.05) lower than those of pathogenic bacteria in the control group due to the organic acids and bacteriocin produced by L. acidophilus SRK30 strain.

Probiotic Potential of Plant-Derived Lactic Acid Bacteria with Antihypertensive Activity (항고혈압 활성을 가진 식물유래 젖산균의 생균제 특성)

  • Lee, Ye-Ram;Son, Young-Jun;Park, Soo-Yun;Jang, Eun-Young;Yoo, Ji-Yeon;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.25 no.6
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    • pp.789-798
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    • 2016
  • Lactic acid bacteria (LAB) are industrially important microorganisms for probiotics. The recent widespread application of LAB for preparation of functional food is attributable to the accumulating scientific evidence showing their beneficial effects on human health. In this study, we isolated and characterized plant-derived LAB that show angiotensin-converting enzyme (ACE) inhibitory and antioxidant activities. The selected strain K2 was isolated from Kimchi, and identified as Lactobacillus plantarum by 16S rRNA gene analysis. The strain grew under static and shaking culture systems. They were also able to grow in different culture conditions like $25^{\circ}C{\sim}37^{\circ}C$ temperature, 4~10 pH range and ~6% NaCl concentration. L. plantarum K2 was highly resistant to acid stress; survival rate of the strain at pH 2.5 and 3 were 80% and 91.6%, respectively. The strain K2 also showed high bile resistance to 0.3% bile bovine and 0.3% bile extract with more than 74% of survival rate. The cell grown on MRS agar plate containing bile extract formed opaque precipitate zones around the colonies, indicating they have bile salt hydrolase activity. The strain showed an inhibitory activity against pathogenic bacteria such as Escherichia coli, Staphylococcus aureus and Listeria monocytogenes; antibacterial activity was probably due to the lactic acid. The K2 strain showed relatively higher autoaggregation values, antihypertensive and antioxidant activities. These results suggest that L. plantarum K2 could be not only applied as a pharmabiotic for human health but also is also starter culture applicable to fermentative products.

Comparison of bacterial communities in leachate from decomposing bovine carcasses

  • Yang, Seung Hak;Ahn, Hee Kwon;Kim, Bong Soo;Chang, Sun Sik;Chung, Ki Yong;Lee, Eun Mi;Ki, Kwang Seok;Kwon, Eung Gi
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.11
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    • pp.1660-1666
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    • 2017
  • Objective: Burial is associated with environmental effects such as the contamination of ground or surface water with biological materials generated during the decomposition process. Therefore, bacterial communities in leachates originating from the decomposing bovine carcasses were investigated. Methods: To understand the process of bovine (Hanwoo) carcass decomposition, we simulated burial using a lab-scale reactor with a volume of $5.15m^3$. Leachate samples from 3 carcasses were collected using a peristaltic pump once a month for a period of 5 months, and bacterial communities in samples were identified by pyrosequencing of the 16S rRNA gene. Results: We obtained a total of 110,442 reads from the triplicate samples of various sampling time points (total of 15 samples), and found that the phylum Firmicutes was dominant at most sampling times. Differences in the bacterial communities at the various time points were observed among the triplicate samples. The bacterial communities sampled at 4 months showed the most different compositions. The genera Pseudomonas and Psychrobacter in the phylum Proteobacteria were dominant in all of the samples obtained after 3 months. Bacillaceae, Clostridium, and Clostridiales were found to be predominant after 4 months in the leachate from one carcass, whereas Planococcaceae was found to be a dominant in samples obtained at the first and second months from the other two carcasses. The results showed that potentially pathogenic microbes such as Clostridium derived from bovine leachate could dominate the soil environment of a burial site. Conclusion: Our results indicated that the composition of bacterial communities in leachates of a decomposing bovine shifted continuously during the experimental period, with significant changes detected after 4 months of burial.

Microbial Floral Dynamics of Chinese Traditional Soybean Paste (Doujiang) and Commercial Soybean Paste

  • Gao, Xiuzhi;Liu, Hui;Yi, Xinxin;Liu, Yiqian;Wang, Xiaodong;Xu, Wensheng;Tong, Qigen;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.12
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    • pp.1717-1725
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    • 2013
  • Traditional soybean paste from Shandong Liangshan and Tianyuan Jiangyuan commercial soybean paste were chosen for analysis and comparison of their bacterial and fungal dynamics using denaturing gel gradient electrophoresis and 16S rRNA gene clone libraries. The bacterial diversity results showed that more than 20 types of bacteria were present in traditional Shandong soybean paste during its fermentation process, whereas only six types of bacteria were present in the commercial soybean paste. The predominant bacteria in the Shandong soybean paste were most closely related to Leuconostoc spp., an uncultured bacterium, Lactococcus lactis, Bacillus licheniformis, Bacillus spp., and Citrobacter freundii. The predominant bacteria in the Tianyuan Jiangyuan soybean paste were most closely related to an uncultured bacterium, Bacillus licheniformis, and an uncultured Leuconostoc spp. The fungal diversity results showed that 10 types of fungi were present in the Shandong soybean paste during the fermentation process, with the predominant fungi being most closely related to Geotrichum spp., an uncultured fungal clone, Aspergillus oryzae, and yeast species. The predominant fungus in the commercial soybean paste was Aspergillus oryzae.

Bacillus oryzicola sp. nov., an Endophytic Bacterium Isolated from the Roots of Rice with Antimicrobial, Plant Growth Promoting, and Systemic Resistance Inducing Activities in Rice

  • Chung, Eu Jin;Hossain, Mohammad Tofajjal;Khan, Ajmal;Kim, Kyung Hyun;Jeon, Che Ok;Chung, Young Ryun
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.152-164
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    • 2015
  • Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and $YC7010^T$, with antimicrobial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension ($10^7cfu/ml$) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC $15859^T$ (99.67%), Bacillus methylotrophicus KACC $13105^T$ (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC $17177^T$ (99.60%), and Bacillus tequilensis KACC $15944^T$ (99.45%). The DNA-DNA relatedness value between strain $YC7010^T$ and the most closely related strain, B. siamensis KACC $15859^T$ was $50.4{\pm}3.5%$, but it was $91.5{\pm}11.0%$ between two strains YC7007 and $YC7010^T$, indicating the same species. The major fatty acids of two strains were anteiso-$C_{15:0}$ and iso $C_{15:0}$. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and $YC7010^T$ represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is $YC7010^T$ (= KACC $18228^T$). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases.

Ecological Characteristics of Actinomycetes from Mercury and Chrome Polluted Soil (수은, 6가크롬 오염토양으로부터 분리된 방선균군의 생태학적 특성)

  • Cho Min-Hye;Han Sang-Mi;Baek Ha-Ju;Whang Kyung-Sook
    • Korean Journal of Environmental Biology
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    • v.24 no.1 s.61
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    • pp.38-45
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    • 2006
  • Ecological characteristics of microbial populations inhabiting heavy metal polluted soil were investigated. The samples were collected from 293 sites around an factory and industry at Gyeoungsangbuk-do. We measured the contents of seven heavy metal elements (Cd, Cu, As, Hg, Pb, $Cr^{6+}$, CN), seven sites have been seriously contaminated by mercury and chrome. A quantitative evaluation of microbial populations in mercury and chrome contaminated soil was examined by using plate count method. Bacterial numbers in polluted soil samples ranged from $7.4X10^5\;to\;9.3X10^7\;cfu\;g^{-1}$, about $10\sim100$ fold less than the count for the unpolluted soil. Moulds were not detected in chrome polluted soil. The log values of actinomycetes of each contaminated soil samples were log ranged from 6.18 to 7.52. The ratio of actinomycetes was similar to unpolluted soil. The investigation showed actinomycetes to be the major microbial population inhabiting the mercury and chrome polluted soil. Thirty-one isolates among the total isolates were examined for antibacterial activity. These isolates were identified based on a phylogenetic analysis using 16S rRNA gene nucleotide sequences, they were categorized in three major phylogenetic groups, belong to the Streptomyces (6 strains), Saccharopolyspora (3 strains), Nocardiodes (1 strain). On the phylogenetic tree, the clade consisting of five isolates were distantly related to all of the established Streptomycetes genera, indicating the possibility as members of new species.

Isolation of Bacillus velezensis SSH100-10 with Antifungal Activity from Korean Traditional Soysauce and Characterization of Its Antifungal Compounds (전통재래 간장으로부터 항진균 활성 B. velezensis SSH100-10의 분리와 그 항진균 물질의 특성 구명)

  • Chang, Mi;Moon, Song Hee;Chang, Hae Choon
    • Food Science and Preservation
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    • v.19 no.5
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    • pp.757-766
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    • 2012
  • The SSH100-10 bacterial strain, which exhibits strong antifungal (anti-mold and anti-yeast) activity, was isolated from traditional korean soysauce aged 100 years. The strain was identified as Bacillus velezensis based on Gram-staining, the biochemical properties and 16S rRNA gene sequence determination. B. velezensis SSH100-10 showed strong proteinase activity and NaCl tolerance, but did not produce enterotoxin. Two-antifungal compounds from B. velezensis SSH100-10 were purified using SPE, preparative HPLC, and reverse phase-HPLC. The purified antifungal compounds were identified as $C_{14}$ and $C_{15}$ iturin through MALDI-TOF-MS and amino acid composition analysis. The stability characteristics of the antifungal compounds after temperature, pH, and enzyme treatments suggested that B. velezensis SSH100-10 produced more than two antifungal compounds; pH-stable $C_{14}$ iturin A and $C_{15}$ iturin A, and unidentified pH-unstable compounds. The results suggested that B. velezensis SSH100-10 can be used in soybean fermentation as a starter. Moreover it has potential as a biopreservative in the food and feed industry and as a biocontrol agent in the field of agriculture.

Characteristic study and isolation of Bacillus subtilis SRCM 101269 for application of cow manure (우분 적용을 위한 Bacillus subtilis SRCM 101269의 분리 및 특성 연구)

  • Jeon, SaeBom;Oh, HyeonHwa;Uhm, Tai-Boong;Cho, Jae-Young;Yang, Hee-Jong;Jeong, Do-Youn
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.74-83
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    • 2016
  • Bacillus subtilis SRCM 101269 having safety and amo gene isolated from Korean traditional fermented food and their investigated characterization to apply the cow manure such as cellulase and xylanase activities, 16S rRNA sequencing, and ability of removal of livestock manure odor. Cow manure application results for the removal of livestock manure odor, the ammonia gas was reduced more than two-folder compared to the control group after 6 days, and reduced to less than 10 ppm after 9 days. In the case of cow manure added fowl droppings and other wood-based mixture components, ammonia gas maintained constant after 3 days of fermentation. However, in the case of sample inoculated B. subtilis SRCM 101269, ammonia gas reduced in course of fermentation time, and concentration of hydrogen sulfide also reduced for 65 ppm. Changes of nitrite concentration according to fermentation time no showed different for cow manure, however nitrite concentration in mixed livestock manure increased when compared to control. And then sulfate concentration in cow manure decreased, and no showed different when compared to the initial fermentation. No apparent change of sulfate concentration in mixed livestock manure detected. Through the previously studies, B. subtilis SRCM 101269 has high potential in industrial application manufacturing the cow manure as removal of livestock manure odor.