• 제목/요약/키워드: Microbial culture

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Composting of Organic Wastes by solid State Fermentation Reactor (Solid State Fermentation Reactor를 이용한 유기성 폐기물의 발효)

  • 홍운표;이신영
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.311-319
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    • 1999
  • Leaves of Aloe vera Linne and bloods of domestic animal were composted in a soild state fermentation reactor (SSFR) by using microbial additive including a bulking and moisture controlling agent. From solid-culture of microbial additive, 10 species of bacteria and 10 species of fungi were isolated and, their enzyme activities including amylase, carboxy methyl cellulase CMCase, lipase and protease were detected. Optimum fermentation conditions of Aloe leaves and domestic animal bloods in SSFR were obtained from the studies of response surface analysis employing microbial additive content, initial moisture content, and fermentation temperature as the independent variables. The optimum conditions for SSFR using Aloe leaves were obtained at 9.45$\pm$73%(w/w) of microbial additives, 62.73$\pm$4.54%(w/w) of initial moisture content and 55.32$\pm$3.14$^{\circ}C$ of fermentation temperature while those for SSFR using domestic animal bloods were obtained at 10.25$\pm$2.04%, 58.68$\pm$4.97% and 57.85$\pm$5.$65^{\circ}C$, respectively. Composting process in SSFR was initially proceeded through fermentation and solid materials were decomposed within 24 hours by maintaining higher moisture level, and maturing and drying steps are followed later. After the fermentation step, the concentrations of solid phase inorganic components were increased while that of organic components were decreased. Also, concentrations of total organic carbon(TOC), peptides, amino acids, polysaccharides, and low fatty acids in water extracts were increased. As fermentation in composting process depends on initial C/N ratios in water extracts of two samples were increased because of increased water-soluble TOC. From these results, it was revealed that solid state fermentation reactor using microbial additives can be used in composting process of organic wastes with broad C/N ratio.

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The microbial diversity analysis of the Korea traditional post-fermented tea (Chungtaejeon) (한국 전통 미생물발효차(청태전)의 미생물 군집분석)

  • Kim, Byung-Hyuk;Jang, Jong-Ok;Kang, Zion;Joa, Jae Ho;Moon, Doo-Gyung
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.170-179
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    • 2017
  • Tea is the most popular beverage in the world. In fact, there are mainly three different kinds of tea (Green tea, black tea, and post-fermented tea). Post-fermented tea is produced by the microbial fermentation process using sun-dried green tea leaves (Camellia sinensis) as the raw material. Chungtaejeon was a traditional tea introduced in the age of the ancient three states and is the only "Ddeok-cha or Don-cha" culture in the world that survived on the southwestern shore of Republic of Korea. In this study, the structures of the bacterial community involved in the production of oriental traditional post-fermented tea (Chungtaejeon) were investigated using 16S rRNA gene analysis. The 16S rRNA gene analysis of dominant microbial bacteria in post-fermented tea confirmed the presence of Pantoea sp., and Klebsiella oxytoca. Phylogenetic analysis suggested that the taxonomic affiliation of the dominant species in the post-fermented tea was ${\gamma}$-proteobacteria. As a result of the microbial community size analysis, it was confirmed that the size of the microbial communities of Chungtaejeon was the largest compared to other teas

Fermentative characteristics of wheat bran direct-fed microbes inoculated with starter culture

  • Kim, Jo Eun;Kim, Ki Hyun;Kim, Kwang-Sik;Kim, Young Hwa;Kim, Dong Woon;Park, Jun-Cheol;Kim, Sam-Chul;Seol, Kuk-Hwan
    • Korean Journal of Agricultural Science
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    • v.43 no.3
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    • pp.387-393
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    • 2016
  • This study was conducted to determine the fermentative characteristics of wheat bran inoculated with a starter culture of direct-fed microbes as a microbial wheat bran (DMWB) feed additive. Wheat bran was prepared with 1% (w/w, 0.5% Lactobacillus plantarum and 0.5% of Saccharomyces cerevisiae) starter culture treatment (TW) or without starter culture as a control (CW). Those were fermented under anaerobic conditions at $30^{\circ}C$ incubation for 3 days. Samples were taken at 0, 1, 2, and 3 days to analyze chemical composition, microbial growth, pH, and organic acid content. Chemical composition was not significantly different between CW and TW (p > 0.05). In TW, the number of lactic acid bacteria and yeast increased during the 3 days of fermentation (p < 0.05) and the population of lactic acid bacteria was significantly higher than in CW (p < 0.05). After 3 days, the number of yeast in TW was $7.50{\pm}0.07log\;CFU/g$, however, no yeast was detected in CW (p < 0.05). The pH values of both wheat bran samples decreased during the 3 days of fermentation (p < 0.05), and TW showed significantly lower pH than CW after 3 days of fermentation (p < 0.05). Contents of lactic acid and acetic acid increased significantly at 3rd day of fermentation in TW. However, no organic acids were generated in CW during testing period. These results suggest that 3 days of fermentation at $37^{\circ}C$ incubation after the inoculation wheat bran with starter culture makes it possible to produce a direct-feed with a high population of lactic acid bacteria at more than $10^{11}CFU/g$.

Change of Insulin-like Growth Factor Gene Expression in Chinese Hamster Ovary Cells Cultured in Serum-free Media

  • Park, Hong-Woo;An, Sung-Kwan;Choe, Tae-Boo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.4
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    • pp.319-324
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    • 2006
  • Although the sera used in animal cell culture media provide the macromolecules, nutrients, hormones, and growth factors necessary to support cell growth, it could also be an obstacle to the production of recombinant proteins in animal cell culture systems used in many sectors of the biotechnology industry. For this reason, many research groups, including our laboratory, have been trying to develop serum-free media (SFM) or serum-supplemented media (SSM) for special or multi-purpose cell lines. The Chinese hamster ovary (CHO) cell, for example, is frequently used to produce proteins and is especially valuable in the large-scale production of pharmaceutically important proteins, yet information about its genome is lacking. Also, SFMs have only been evaluated by comparing growth patterns for cells grown in SFMs with those grown in SSM or by measuring the titer of the target protein obtained from cells grown in each type of medium. These are not reliable methods of obtaining the type of information needed to determine whether an SFM should be replaced with an SSM. We carried out a cDNA microarray analysis to evaluate MED-3, an SFM developed in our laboratory, as a CHO culture medium When CHO cells were cultured in MED-3 instead of an SSM, several genes associated with cell growth were down-regulated, although this change diminished over time. We found that the insulin-like growth factor (IGF) gene was representative of the proteins that were down-regulated in cells cultured in MED-3. When several key supplements - including insulin, transferrin, ethanolamine, and selenium - were removed from MED-3, the IGF expression was consistently down- regulated and cell growth decreased proportionately. Based on these results, we concluded that when an SFM is used as a culture medium, it is important to supplement it with substances that can help the cells maintain a high level of IGF expression. The data presented in this study, therefore, might provide useful information for the design and development of SFM or SSM, as well as for the design of genome-based studies of CHO cells to determine how they can be used optimally for protein production in pharmaceutical and biomedical research.

Overexpression of Thermoalkalophilic Lipase from Bacillus stearothermophilus L1 in Saccharomyces cerevisiae

  • Ahn, Jung-Oh;Jang, Hyung-Wook;Lee, Hong-Weon;Choi, Eui-Sung;Haam, Seung-Joo;Oh, Tae-Kwang;Jung, Joon-Ki
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.451-456
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    • 2003
  • An expression vector system was developed for the secretory production of recombinant Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae. The mature L1 lipase gene was fused to ${\alpha}-amylase$ signal sequence from Aspergillus oryzae for the effective secretion into the culture broth and the expression was controlled under GAL10 (the gene coding UDP-galactose epimerase of S. cerevisiae) promoter. S. cerevisiae harboring the resulting plasmid successfully secreted L1 lipase into the culture broth. To examine an optimum condition for L1 lipase expression in the fed-batch culture, L1 lipase expression was induced at three different growth phases (early, mid, and late-exponential growth phases). Maximum product on of L1 lipase (1,254,000 U/l, corresponding to 0.65/1) was found when the culture was induced at an early growth phase. Secreted recombinant L1 lipase was purified only through CM-Sepharose chromatography, and the purified enzyme showed 1,963 U/mg of specific activity and thermoalkalophilic properties similar to those reported for the enzyme expressed in Escherichia coli.

Optimization of Culture Conditions for Erythritol Production by Torula sp.

  • Kim, Kyung-Ah;Noh, Bohg-Soo;Lee, Jung-Kul;Kim, Sang-Yong;Park, Yong-Cheol;Oh, Deok-Kun
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.69-74
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    • 2000
  • The medium for erythritol production by Torula sp. in a 500-ml baffled flask was optimized to be 300 g/I sucrose, 10 g/I yeast extract, 3 g/I $KH_2PO_4$, and 10 mg/I $CuSO_4{\cdot}5H_2O{\;}at{\;}34^{\circ}C$ with initial pH of 5.5. Using this optimal medium, erythritol of 166 g/I was obtained after 140 h of cultivation, corresponding to 55.3% of the erythritol yield from sucrose with a productivity of 1.11 g/I/h. Optimal concentrations of carbbon and nitrogen sources in a fermentor were higher than that in a flask due to the higher oxygen supply of the fermentor. Employing the medium containing 300 g/I or 400 g/I sucrose for the determination of optimal C/N ratio, the C/N ratio was found to be more important than the nitrogen concentration for effective erythritol production, The optimal ratio of yeast extract to sucrose (g/g) was 20. The yield and productivity of erythritol were maximal in the medium containing 400 g/I sucrose and 20 g/I yeast extract. when dissolved oxygen in the culture was increased, the cell mass increased but the erythritol production was manimal in the range of 5 to 10% of dissolved oxygen. Under the optimal the rane of 5 to 10% of dissolved oxygen. Under the optimal culture condition of the fermentor, a final erythritol concentration of 200 gI was obtained after 120 h with a yield of 50% and the productivity was 1.67 g/I/h. The yield was the highest among erythritol-producting microorganisms

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Effect of Glasswort (Salicornia herbacea L.) on Microbial Community Variations in the Vinegar-making Process and Vinegar Characteristics

  • Seo, Ha-Na;Jeon, Bo-Young;Yun, A-Ram;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.20 no.9
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    • pp.1322-1330
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    • 2010
  • Three types of nuruk were made from rice, wheat, and a rice-glasswort (6:4) mixture. Nuruk, makgeolli, and vinegar were manufactured with rice nuruk (RN), wheat nuruk (WN), and rice-glasswort nuruk (RGN). The variable region of 18S or 16S rDNA amplified with genomic DNA extracted directly from nuruk-, makgeolli-, and vinegar-making cultures was analyzed via temperature gradient gel electrophoresis (TGGE). The sequence of the 18S rDNA variable region extracted from the TGGE gel for nuruk was 99% homologous with Aspergillus sp. and that for the makgeolli-making culture was 99% homologous with Saccharomyces sp. and Saccharomycodes sp. The sequence of the 16S rDNA variable region extracted from TGGE gel for the vinegar-making culture was 98% homologous, primarily with the Acetobacter sp. The eukaryotic and prokaryotic diversities in the nuruk-, makgeolli-, and vinegar-making cultures was not significantly altered by the addition of glasswort. Prokaryotic diversity was higher than eukaryotic diversity in the nuruk, but eukaryotic diversity was higher than prokaryotic diversity in the makgeolli-making culture, on the basis of the TGGE patterns. No 18S rDNA was amplified from the DNA extracted from the vinegar-making culture. The diversity of the microbial community in the process from nuruk to vinegar was slightly affected by the type of raw material utilized for nuruk-making. The saccharifying activity and ethanol productivity of nuruk, polyphenol content in makgeolli, and acetic acid and polyphenol content in the vinegar were increased as a result of the addition of glasswort. In conclusion, the glasswort may be not simply an activator for the growth of microorganisms during the fermentation of nuruk, makgeolli, or vinegar, but also a nutritional supplement that improves the quality of vinegar.

Nitrogen Removals according to Aeration/Non-aeration Periods in the Intermittent Aeration Reactor and Analysis of Microbial Community (간헐포기공정에서 포기/비포기 구간에 따른 질소제거 및 미생물 군집분석)

  • Choi, Moon-Su;Lee, Tae-Jin
    • Journal of Korean Society of Environmental Engineers
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    • v.36 no.1
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    • pp.42-48
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    • 2014
  • In this study, variations of the organic and nitrogenous compounds in wastewater were investigated in a single reactor with intermittent aeration. Over 90% of organic and nitrogen removals are accomplished with C/N ratio of 3 : 1 and 20/20 min of aeration/non-aeration period. Longer non-aeration period on the aeration/non-aeration cycle showed more stable nitrogen removal, showing various microbial community in the reactor. From PCR-DGGE analysis, it is conclusive that Dysgonomonas mossii strain Melo40, Eubacterium sp. oral clone JN088, Uncultured bacterium clone SPESB2_718, and Bacterium enrichment culture clone LE are related with the organics and nitrogen oxidation. Uncultured Acidobacteria bacterium clone AKYG487, Lactobacillus harbinensis strain FQ003, Erythrobacter litoralis strain Gi-3, Phytobacter diazotrophicus strain Ls8, and Mycobacterium sp. enrichment culture clone GE10037biofNNA are distinctly appeared under denitrification condition.

Effect of Multifaceted Interventions for Ward Nurses on the Storage, Conditions, and Transportation of Specimens for Microbial Culture (병동 간호사 대상의 다각적 중재가 배양검사 검체의 보관과 상태 및 운송에 미치는 영향)

  • Cho, Min Jung;Jeong, Jae Sim;Kim, Yoon Hee
    • Journal of Korean Biological Nursing Science
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    • v.24 no.2
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    • pp.95-103
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    • 2022
  • Purpose: The purpose of this study was to provide multifaceted interventions for nurses and to confirm changes in their knowledge, perception, and actual practice on the adequate storage, conditions, and transportation of culture specimens. Methods: A one-group pretest-posttest experimental design was conducted with 41 nurses in two general wards of a tertiary acute care hospital in Seoul. Multifaceted interventions including education, feedback, posting guidelines and reminders, and improvement in specimen management accessibility were provided from May 2019 to January 2020. Outcomes were measured before and after the interventions. Knowledge and perception of the nurses were evaluated using self-reported questionnaires and actual practice by observation. Results: After the interventions, the average knowledge score on transportation time was significantly increased (Z= -4.89, p< .001). However, the knowledge score on storage methods was not significantly increased. The perception score was significantly increased (t= -3.19, p= .003). The proportion of specimen storage times, places, and conditions managed properly was significantly increased from 43.0% (46/107) to 77.1% (84/109) (p< .001). The average transportation time of blood samples to the laboratory significantly decreased from 3 hours 36 minutes (± 1 hour 52 minutes) to 3 hours 1 minute (± 1 hour 41 minutes) (t= 2.51, p= .013). The percentage of blood culture specimens arriving within 2 hours was increased significantly from 22.9% to 39.2% (χ2= 6.90, p= .009). Conclusion: The interventions were effective. However, some specimens remained in the ward longer than expected after the interventions. This requires further interventions.