• Journal of Microbiology and Biotechnology
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• v.32 no.3
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• pp.341-347
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• 2022

In this study, the bacterial community of galchi-baechu kimchi was determined using culture-based and culture-independent techniques (next generation sequencing:NGS), and showed discrepancies between results. Weissella koreensis and Pediococcus inopinatus were the dominant species according to the NGS results, while Bacillus species and P. inopinatus were dominant in the culture-dependent analysis. To identify safe starter candidates, sixty-five Bacillus strains isolated from galchi-baechu kimchi using culture-dependent methods were evaluated for their antibiotic resistance, presence of toxin genes, and hemolytic activity. Strains were then assessed for salt tolerance and protease and lipase activity. As a result, four strains-B. safensis GN5_10, B. subtilis GN5_19, B. velezensis GN5_25, and B. velezensis GT8-were selected as safe starter candidates for use in fermented foods.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.171-173
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• 2003

Suspension culture in serum-free medium is important for the efficient large-scale culture of anchorage-dependent cells that are utilized to produce therapeutic recombinant protein(e.g., insulin, antibody, vaccine) and virus vector for therapeutic gene transfer. We developed a novel method for the suspension culture of anchorage-dependent animal cells in serum-free medium using biodegradable polymer nanospheres in this study. Poly(lactic-co-glycolic acid) (PLGA) polymer nanospheres (433nm in average diameter) were used to the culture of human embryonic kidney 293 cells in serum-free medium in stirred suspension bioreactors. The use of PLGA nanospheres promoted the aggregate formation and cell growth (3.8-fold versus 1.8-fold growth), compared to culture without nanospheres. Adaptation of the anchorage-dependent cells to suspension culture or serum-free medium is time-consuming and costly. In contrast, the culture method developed in our study does not require the adaptation process. This method may be useful for the large-scale suspension culture of various types of anchorage-dependent animal cells in serum-free medium.

• Journal of Microbiology and Biotechnology
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• v.8 no.5
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• pp.461-465
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• 1998

The suspension culture of an anchorage-dependent cell line (Bok-l) from Bombina orientalis was successful in respects of cost and efficiency. The amount of cells obtained from the suspension culture was almost equivalent to that from the anchorage-dependent culture. This result shows the possibility of suspension culture for scale-up. The cells in suspension produced an antibacterial peptide as much as anchorage-dependent cells did. The cell growth ($6.0\times10^6cells/m\ell$) and viability (>80%) at 10 rpm were higher than that at 0 rpm ($1.9\times10^6cells/m\ell$, 65~80%) and 30 rpm ($1.8\times10^6cells/m\ell$ 40~76%). The size of cells became smaller at the agitation rate of 30 rpm. The antibacterial activities of cell extracts from suspension cultured cells were confirmed against gram-negative and gram-positive bacteria by the inhibition zone assay and the liquid growth inhibition assay.

• Journal of Microbiology and Biotechnology
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• v.28 no.1
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• pp.32-39
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• 2018

Samples of Laru (a fermentation starter) obtained from the upper part of Borneo Island were analyzed for their lactic acid bacteria (LAB) and fungal diversity using both a culture-independent method (PCR-DGGE) and culture-dependent methods (SDS-PAGE and MALDI-TOF MS). Pediococcus pentosaceus, Lactobacillus brevis, Saccharomycopsis fibuligera, Hyphopichia burtonii, and Kodamaea ohmeri were detected by all three methods. In addition, Weissella cibaria, Weissella paramesenteroides, Leuconostoc citreum, Leuconostoc mesenteroides, Lactococcus lactis, Rhizopus oryzae/Amylomyces rouxii, Mucor indicus, and Candida intermedia were detected by PCR-DGGE. In contrast, Lactobacillus fermentum, Lactobacillus plantarum, Pichia anomala, Candida parapsilosis, and Candida orthopsilosis were detected only by the culture-dependent methods. Our results indicate that the culture-independent method can be used to determine whether multiple laru samples originated from the same manufacturing region; however, using the culture-independent and the two culture-dependent approaches in combination provides a more comprehensive overview of the laru microbiota.

• Journal of Korean Society on Water Environment
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• v.24 no.2
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• pp.195-200
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• 2008

To analyze the riverine microbial community structure, genetic fingerprints and ecological indexes such as species abundances, diversity, evenness, dominance of targeted rivers in Gyeonggi Province were acquired and evaluated using terminal restriction fragment length polymorphism (T-RFLP) technique. Genetic fingerprinting technique such as T-RFLP, which is able to show the microbial community clearly unlike traditional culture-dependent techniques, was thought to be useful to analyse the riverine microbial ecosystem under various factors. Riverine ecosystem evaluation using visible organisms would give biased results with time, targeted organism and researcher. But, T-RFLP, which can exclude the subjected biases such as culture condition and identification, would be an option to understand natural ecosystem by including the microorganisms that defy culture but perform important functions.

• YAKHAK HOEJI
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• v.35 no.3
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• pp.245-251
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• 1991

Selective culture of actinomycetes from soil microbes and their antibiotic producing characters by agar-disk method were examined. Some of the organisms which produced antibiotics on agar disk did not produce antibiotics in liquid culture. Further examination indicated that production of antibiotic was dependent on the composition of medium. Many streptomycestes produced antibacterial substances in tryptic soy broth but others produced antifungal antibiotics in V-8 broth. Production of antibacterial substances by Streptomyces sp. was also dependent on the medium composition.

• Journal of Animal Science and Technology
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• v.63 no.6
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• pp.1423-1432
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• 2021

To elucidate the role and mechanism of microbes, we combined culture-dependent and culture-independent approaches to investigate differences in gut bacterial composition between sows and weaned pigs. Under anaerobic conditions, several nonselective and selective media were used for isolation from fecal samples. All isolated bacteria were identified and classified through 16S rRNA sequencing, and the microbiota composition of the fecal samples was analyzed by metagenomics using next generation sequencing (NGS) technology. A total of 278 and 149 colonies were acquired from the sow and weaned pig fecal samples, respectively. Culturomics analysis revealed that diverse bacterial genus and species belonged to Firmicutes, Actinobacteria, Proteobacteria, and Bacteroidetes were isolated from sow and weaned pigs. When comparing culture-dependent and culture-independent analyses, 191 bacterial species and 2 archaeal bacterial species were detected through culture-independent analysis, and a total of 23 bacteria were isolated through a culture-dependent approach, of which 65% were not detected by metagenomics. In conclusion, culturomics and metagenomics should be properly combined to fully understand the intestinal microbiota, and livestock-derived microbial resources should be informed by culturomic approaches to understand and utilize the mechanism of host-microbe interactions.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.80-104
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• 1996

The development of routine techniques for the isolation and in vitro maintenance of conducting airway epithelial cells in a differentiated state provides an ideal model to study the factors involved in the regulation of the expression of mucocilicary differentiation. Several key factors and conditions have been identified. These factors and conditions include the use of biphasic culture technique to achieve mucociliary differentiation and the use of such stimulators, the thickness of collagen gel substratum, the calcium level, and vitamin A, and such inhibitors, the growth factors EGF and insulin, and steroid hormones, for mucous cell differentiation. Using the defined culture medium, the life cycle of the mucous cell population in vitro was investigated. It was demonstrated that the majority of the mucous cell population in primary cultures is not involved in DNA replication. However, the mucous cell type is capable of self-renewal in culture and this reproduction is vitamin A dependent. furthermore, differentiation from non-mucous cell type to mucous cell type can be demonstrated by adding back a positive regulator such as vitamin A to the “starved” culture. Cell kinetics data suggest that vitamin A-dependent mucous cell differentiation in culture is a DNA replication-independent process and the process is inhibited by TGF-${\beta}$1.

• KSBB Journal
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• v.6 no.4
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• pp.389-394
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• 1991

Emulsan is an extracellular emulsifying agent produced by the hydrocarbon-degrading Acinetobacter species RAG-1. In this study emulsan production of Acinetobacter calcpaceticus RAG-1 was investigated under various culture modes such as batch, fed-batch, membrane cell recycle, and continuous culture. The productions of emulsan under both ethanol-sufficient fed-batch and membrane cell recycle cultures were all 15.0U/ml, which was 53% increase in emulsan activity compared to that of pH controlled batch culture. Emulsan production was found to be strongly dependent on the residual ethanol concentration. In continuous culture the emulsan productivity increased with dilution rate.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.4
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• pp.447-453
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• 2015

We determined the optimal culture conditions for obtaining the maximum number of intestinal bacteria from the olive flounder Paralichthys olivaceus, and studied bacterial diversity using both culture-dependent and culture-independent methods. Using six culture conditions, mean bacterial numbers were greater than $10^6$ per gram of gut mucus, regardless of the medium. However, the bacterial diversity, based on colony morphology, appeared much higher on Marine agar (MA) and Zobell 2216 agar than on other media. We found eight and 17 cultured bacterial phylotypes with 99% minimum similarity in gut mucus grown on MA and tryptic soy agar, respectively. Furthermore, we used genomic DNA extracted from gut mucus to generate 78 random clones, which were grouped into 25 phylotypes. Of these, six were affiliated with Firmicutes, Actinobacteria, and Verrucomicrobia, and were not found using our culture-dependent methods. Consequently, we believe that Marine agar and Zobell 2216 agar are optimal media for culturing diverse intestinal microbes; we also discovered several novel sequences not previously recognized as part of the gut microbiota of olive flounder.