• Title/Summary/Keyword: l6s rRNA

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Screening of High Temperature-Tolerant Oleaginous Diatoms

  • Zhang, Lingxiang;Hu, Fan;Wan, Xiu;Pan, Yufang;Hu, Hanhua
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.1072-1081
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    • 2020
  • Screening suitable strains with high temperature adaptability is of great importance for reducing the cost of temperature control in microalgae cultivation, especially in summer. To obtain high temperature-tolerant diatoms, water samples were collected in summer from 7 different regions of China across the Northeast, North and East. A total of 731 water samples was collected and from them 131 diatom strains were isolated and identified based on the 18S rRNA sequences. Forty-nine strains out of the 131 diatoms could survive at 30℃, and 6 strains with relatively high biomass and lipid content at high temperature were selected and were found to be able to grow at 35℃. Cyclotella sp. HB162 had the highest dry biomass of 0.46 g/l and relatively high triacylglycerol (TAG) content of 237.4 mg/g dry biomass. The highest TAG content of 246.4 mg/g dry biomass was obtained in Fistulifera sp. HB236, while Nitzschia palea HB170 had high dry biomass (0.33 g/l) but relatively low TAG content (105.9 mg/g dry biomass). N. palea HB170 and Fistulifera sp. HB236 presented relatively stable growth rates and lipid yields under fluctuating temperatures ranging from 28 to 35℃, while Cyclotella HB162 maintained high lipid yield at temperatures below 25℃. The percentage of saturated fatty acids and monounsaturated fatty acids in all the 6 strains was 84-91% in total lipids and 90-94% in TAGs, which makes them the ideal feedstock for biodiesel.

Changes in physicochemical property and lactic acid bacterial community during kimchi fermentation at different temperatures

  • Lee, Hee Yul;Haque, Md. Azizul;Cho, Kye Man
    • Journal of Applied Biological Chemistry
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    • v.63 no.4
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    • pp.429-437
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    • 2020
  • This study aimed to investigate the change in physicochemical properties and lactic acid bacterial communities during kimchi fermentation at different temperatures (8, 15, and 25 ℃) using two molecular genetics approaches, multiplex polymerase chain reaction and 16S rRNA gene sequencing. The pH during fermentation at 8, 15, and 25 ℃ decreased from 6.17 on the initial fermentation day to 3.92, 3.79, and 3.48 after 54, 30, and 24 days of fermentation, respectively, while the acidity increased from 0.24% to 1.12, 1.35, and 1.54%, respectively. In particular, the levels of lactic acid increased from 3.74 g/L on the initial day (day 0) to 14.43, 20.60, and 27.69 g/L during the fermentation after 24, 18, and 12 days at 8, 15, and 25 ℃, respectively, after that the lactic acid concentrations decreased slowly. The predominance of lactic acid bacteria (LAB) in the fermented kimchi was dependent on fermentation stage and temperature: Lactobacillus sakei appeared during the initial stage and Leuconsotoc mesenteroides was observed during the optimum-ripening stage at 8, 15, and 25 ℃. Lac. sakei and Lactobacillus plantarum grew rapidly in kimchi produced at 8, 15, and 25 ℃. In addition, Weissella koreensis first appeared at days 12, 9, and 6 at 8, 15, and 25 ℃ of fermentation, respectively. This result suggests that LAB population dynamics are rather sensitive to environmental conditions, such as pH, acidity, salinity, temperature, and chemical factors including free sugar and organic acids.

Effects of Acarbose Addition on Ruminal Bacterial Microbiota, Lipopolysaccharide Levels and Fermentation Characteristics In vitro

  • Yin, Yu-Yang;Liu, Yu-Jie;Zhu, Wei-Yun;Mao, Sheng-Yong
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.12
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    • pp.1726-1735
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    • 2014
  • This study investigated the effects of acarbose addition on changes in ruminal fermentation characteristics and the composition of the ruminal bacterial community in vitro using batch cultures. Rumen fluid was collected from the rumens of three cannulated Holstein cattle fed forage ad libitum that was supplemented with 6 kg of concentrate. The batch cultures consisted of 8 mL of strained rumen fluid in 40 mL of an anaerobic buffer containing 0.49 g of corn grain, 0.21 g of soybean meal, 0.15 g of alfalfa and 0.15g of Leymus chinensis. Acarbose was added to incubation bottles to achieve final concentrations of 0.1, 0.2, and 0.4 mg/mL. After incubation for 24 h, the addition of acarbose linearly decreased (p<0.05) the total gas production and the concentrations of acetate, propionate, butyrate, total volatile fatty acids, lactate and lipopolysaccharide (LPS). It also linearly increased (p<0.05) the ratio of acetate to propionate, the concentrations of isovalerate, valerate and ammonia-nitrogen and the pH value compared with the control. Pyrosequencing of the 16S rRNA gene showed that the addition of acarbose decreased (p<0.05) the proportion of Firmicutes and Proteobacteria and increased (p<0.05) the percentage of Bacteroidetes, Fibrobacteres, and Synergistetes compared with the control. A principal coordinates analysis plot based on unweighted UniFrac values and molecular variance analysis revealed that the structure of the ruminal bacterial communities in the control was different to that of the ruminal microbiota in the acarbose group. In conclusion, acarbose addition can affect the composition of the ruminal microbial community and may be potentially useful for preventing the occurrence of ruminal acidosis and the accumulation of LPS in the rumen.

Isolation of Exopolysaccharide-Producing Lactic Acid Bacteria from Pa-Kimchi and Characterization of Exopolysaccharides

  • Yun Ji Kang;Tae Jin Kim;Min Jae Kim;Ji Yeon Yoo;Jeong Hwan Kim
    • Microbiology and Biotechnology Letters
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    • v.51 no.2
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    • pp.157-166
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    • 2023
  • Three lactic acid bacteria (LAB) producing exopolysaccharides (EPSs) were isolated from Pa (green onion)-kimchi, and identified as Weissella confusa (SKP 173), Weissella cibaria (SKP 182), and Leuconostoc citreum (SKP 281), respectively by 16S rRNA gene sequencing. The yields of EPS were 21.27, 18.53, and 15.4 g/l for EPS from SKP 173, 182, and 281, respectively when grown in MRS broth containing sucrose (5%, w/v). Total sugar contents were 64.39, 62.84, and 65.16% (w/w) for EPS from SKP 173, 182, and 281, respectively while the protein contents were 0.33, 0.31, and 0.25% (w/w), respectively. EPSs from W. confusa SKP 173 and W. cibaria SKP 182 contained glucose only but EPS from L. citreum SKP 281 contained glucose and glucitol. Viscosities of the 2% (w/w) freeze-dried EPS solution were 9.60, 8.00, and 8.20 centipoise (cP) for EPS from SKP 173, 182, and 281, respectively. Viscosities of culture grown in MRS broth with 5% sucrose (no glucose) were 92.98, 57.19, and 18.8 cP, respectively. The average molecular weights of EPSs were larger than 2 × 107 Da. Fourier transform infrared spectroscopy (FT-IR) analyses of EPSs showed typical carbohydrate peaks, suggesting that the EPSs consisted of pyranose saccharides with α-(1,6) and α-(1,3) glycosidic linkages. L. citreim SKP 281 was used as the starter for yogurt fermentation, and EPS production was confirmed.

Improved Viability and Proteome Analysis of Lactobacillus fermentum KLB12 upon Heat Stress (Lactobacillus fermentum KLB12의 열 전처리에 따른 열 스트레스 내성 증진 및 프로테옴 변화)

  • 김주현;박미영;김승철;윤현식;소재성
    • KSBB Journal
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    • v.18 no.4
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    • pp.294-300
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    • 2003
  • In the previous study, we have isolated several vaginal lactobacilli from Korean woman and selected one of them (KLB12) for further study, which was indentified as Lactobacillus fermentum by sequence analysis of 16S rRNA gene. Formulated L. fermentum KLB12 can be used for ecological treatment of bacterial vaginosis. For pharmaceutical formulation, the spray-drying process is required where stress such as high temperature is routinely applied. In this study, we found that heat stress at 60$^{\circ}C$ for 20∼30min reduced the viable cell population of KLB12 by 10$\sub$6/~10$\sub$9/. However, adaptation of KLB12 cells at 52$^{\circ}C$ made them more thermotolerant upon exposure to 60$^{\circ}C$. The level of thermal protection in RSM (reconstituted skim milk) by adaptation in acid (pH 5), cold (4$^{\circ}C$), ethanol (3%), NaCI (0.3M) was also examined. Although not as efficient as the homologous stress, adaptations in both cold and NaCI gave considerable cross protection against heat stress. When chloramphenicol was added during heat adaptation, adaptation effect was abolished. This suggests that de novo protein synthesis is necessary during the adaptation process. Important changes in proteome during heat adaptation was examined with two-dimensional gel electrophoresis.

Phylogentic Position, Pigment Content and Optimal Growth Condition of the Unicellular Hydrogen-Producing Cyanobacterial Strains from Korean Coasts (한국 연안산 단세포성 수소생산 남세균 종주들의 분류계통, 색소함량 및 최적성장 환경)

  • PARK, JONG-WOO;KIM, JU HEE;CHO, AE-RA;JUNG, YUN-DUK;KIM, PYOUNG JOONG;KIM, HYUNG-SEOP;YIH, WONHO
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.20 no.3
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    • pp.131-140
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    • 2015
  • To set up unicellular cyanobacterial strains with photo-biological $H_2$ production potential, live samples were repeatedly collected from 68 stations in the coastal zone of Korea for the four years since 2005. Among 77 cyanobacterial strains established six (KNU strains, CB-MAL002, 026, 031, 054, 055 and 058) were finally chosen as the excellent strains for $H_2$ production with $H_2$ accumulation over 0.15 mL $H_2\;mL^{-1}$ under general basic $H_2$ production conditions as well as positive $H_2$ production for more than 60 hr. To explore optimum procedures for higher $H_2$ production efficiency of the six cyanobacterial strains, the inter-strain differences in the growth rate under the gradients of water temperature and salinity were investigated. The maximum daily growth rates of the six strains ranged from 1.78 to 2.08, and all of them exhibited $N_2-fixation$ ability. Based on the similarity of the 16S rRNA sequences, all the test strains were quite close to Cyanothece sp. ATCC51142 (99%). The six strains, however, were grouped into separate clades from strain ATCC51142 in the molecular phylogeny diagram. Chlorophyll- a content was 3.4~7.8% of the total dried weight, and the phycoerythrin and phycocyanin contents were half of those in the Atlantic strain, Synechococcus sp. Miami BG03511. The growth of the six strains was significantly suppressed at temperatures above the optimal range, $30{\sim}35^{\circ}C$, to be nearly stopped at $40^{\circ}C$. The growth was not inhibited by high salinities of 30 psu salinity in all the strains while strain CB055 maintained its high growth rate at low salinities down to 15 psu. The euryhaline strains like CB055 might support massive biotechnological cultivation systems using natural basal seawater in temperate latitudes. base seawater. The biological and ecophysiological characteristics of the test strains may contribute to designing the optimal procedures for photo-biological $H_2$ production by unicellular cyanobacteria.

Optimization of Soymilk Fermentation by the Protease-producing Lactobacillus paracasei (Protease를 생산하는 Lactobacillus paracasei의 분리와 이를 이용한 두유 발효 최적화)

  • Lee, Sulhee;Jang, Dong-Hun;Choi, Hyuk Jun;Park, Young-Seo
    • Korean Journal of Food Science and Technology
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    • v.45 no.5
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    • pp.571-577
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    • 2013
  • Our aim was to ferment soymilk using lactic acid bacteria that showed protease activity and to optimize the condition for fermentation. In total, 108 strains of protease-producing lactic acid bacteria were isolated from various fermented foods such as kimchi and jeotgal, and among them, 29 strains displaying the highest protease activity were selected for further study. From these 29 strains, strain MK1, whose protease activity was 126 $mU/mL{\cdot}min$, was selected as the optimal fermentation strain owing to its high ability to digest soymilk protein. It was henceforth labeled as Lactobacillus paracasei MK1. The optimum conditions for the fermentation of soymilk by using L. paracasei MK1 were determined to be as follows: 30 h of fermentation time at a temperature of $30^{\circ}C$, and at a pH of 6.0 in the initial growth medium.

Characteristic study and optimization of culture conditions for Bacillus amyloliquefaciens SRCM 100731 as probiotic resource for companion animal (Bacillus amyloliquefaciens SRCM 100731의 반려 동물용 프로바이오틱스 소재로서의 특성 규명 및 배양 조건 최적화)

  • Ryu, Myeong Seon;Yang, Hee-Jong;Jeong, Su-Ji;Seo, Ji Won;Ha, Gwangsu;Jeong, Seong-Yeop;Jeong, Do-Youn
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.384-397
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    • 2018
  • The aim of this study is to screen the strains of Bacillus spp. possessing safety, probiotic activity, and so on, which can be utilized as probiotic resource for using the feed and supplement food of companion animal. About 300 isolates were isolated from traditional Korean sauces, four isolates that did not have or produce the six kinds of B. cereus type vomiting and diarrhea toxin genes, ${\beta}$-hemolytic, and three kinds of carcinogenic enzymes were selected. Antibiotic gene retention, cell surface hydrophobicity, antibiotic sensitivity, and glucose utilization were analyzed for four isolates, and finally SRCM 100731 was selected. SRCM 100731 was named as Bacillus amyloliquefaciens SRCM 100731 16S rRNA sequencing analysis, and carried out optimization of cell growth for industrial applications such as pet food and feed. The effects of 14 different components on cell growth were investigated and three significant positive factors, molasses, sodium chloride, and potassium chloride were selected as the main factors based on a Plackett-Burman design. In order to find out optimal concentration on each constituent, we carried out central composite design. The predicted optimized concentrations were 7% molasses, 1.1% sodium chloride, 0.5% potassium chloride. Finally, an overall about 7-fold increase in dry cell weight yield ($12.6625{\pm}0.0658g/L$) was achieved using the optimized medium compared with the non-optimized medium ($1.8273{\pm}0.0214g/L$). This research is expected to be highly utilized in the growing pet industry by establishing optimal cultivation conditions for industrial application as well as screening Bacillus amyloliquefaciens SRCM 100731 as probiotic resource for companion animal.

Cr(VI) Resistance and Removal by Indigenous Bacteria Isolated from Chromium-Contaminated Soil

  • Long, Dongyan;Tang, Xianjin;Cai, Kuan;Chen, Guangcun;Shen, Chaofeng;Shi, Jiyan;Chen, Linggui;Chen, Yingxu
    • Journal of Microbiology and Biotechnology
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    • v.23 no.8
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    • pp.1123-1132
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    • 2013
  • The removal of toxic Cr(VI) by microorganisms is a promising approach for Cr(VI) pollution remediation. In the present study, four indigenous bacteria, named LY1, LY2, LY6, and LY7, were isolated from Cr(VI)-contaminated soil. Among the four Cr(VI)-resistant isolates, strain LY6 displayed the highest Cr(VI)-removing ability, with 100 mg/l Cr(VI) being completely removed within 144 h. It could effectively remove Cr(VI) over a wide pH range from 5.5 to 9.5, with the optimal pH of 8.5. The amount of Cr(VI) removed increased with initial Cr(VI) concentration. Data from the time-course analysis of Cr(VI) removal by strain LY6 followed first-order kinetics. Based on the 16S rRNA gene sequence, strain LY6 was identified as Pseudochrobactrum asaccharolyticum, a species that had never been reported for Cr(VI) removal before. Transmission electron microscopy and energy dispersive X-ray spectroscopy analysis further confirmed that strain LY6 could accumulate chromium within the cell while conducting Cr(VI) removal. The results suggested that the indigenous bacterial strain LY6 would be a new candidate for potential application in Cr(VI) pollution bioremediation.

Characterization of Agarase Produced from the Isolated Marine Bacterium Marinomonas sp. SH-2 (해양성 Marinomonas sp. SH-2 균주가 생성하는 agarase의 분리 및 특성조사)

  • Jo, Jeong-Gwon;Lee, Sol-Ji;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.26 no.2
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    • pp.198-203
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    • 2016
  • This study aimed to isolate a novel agarase-producing marine bacterium and characterize its agarase, as agarases are known to produce biofunctional agarooligosaccharides or neo-agarooligosaccharides. A novel agar-degrading bacterium, SH-2, was isolated from the seawater of Namhae in Gyeongnam Province, Korea, and cultured in Marine agar 2216 medium. The 16S rRNA gene sequence represented 99% identity with that of the members of the Marinomonas genus; hence, the isolated bacterium was named Marinomonas sp. SH-2. The crude agarase was prepared from a culture medium of Marinomonas. sp SH-2, and exhibited maximum agarase activity at 170.2 units/l. The optimum conditions were pH 6.0 and 30℃ in 20 mM Tris-HCl buffer. The agarase activity of the bacterium was highly elevated from 20℃(42% relative activity) to 30℃(100%), and 82% activity was shown at 40℃. Its relative activities were less than 40% at over 40℃ after a 0.5 hr exposure. Relative activity was 100% at pH 6.0, while it was 72% and 48% at pH 5.0 and pH 7.0, respectively. The enzyme from Marinomonas sp. SH-2 degraded agarose to neoagarohexaose and neoagarotetraose, indicating that the enzyme is β-agarase. Thus, Marinomonas sp. SH-2 and its enzyme could be practical for applications in food, cosmetic, and medical research.