• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1559-1565
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• 2014

Cellulase and xylanase are main hydrolysis enzymes for the degradation of cellulosic and hemicellulosic biomass, respectively. In this study, our aim was to develop and test the efficacy of a rapid, high-throughput method to screen hydrolytic-enzyme-producing microbes. To accomplish this, we modified the 3,5-dinitrosalicylic acid (DNS) method for microwell plate-based screening. Targeted microbial samples were initially cultured on agar plates with both cellulose and xylan as substrates. Then, isolated colonies were subcultured in broth media containing yeast extract and either cellulose or xylan. The supernatants of the culture broth were tested with our modified DNS screening method in a 96-microwell plate, with a $200{\mu}l$ total reaction volume. In addition, the stability and reliability of glucose and xylose standards, which were used to determine the enzymatic activity, were studied at $100^{\circ}C$ for different time intervals in a dry oven. It was concluded that the minimum incubation time required for stable color development of the standard solution is 20 min. With this technique, we successfully screened 21 and 31 cellulase- and xylanase-producing strains, respectively, in a single experimental trial. Among the identified strains, 19 showed both cellulose and xylan hydrolyzing activities. These microbes can be applied to bioethanol production from cellulosic and hemicellulosic biomass.

• Journal of Microbiology and Biotechnology
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• v.24 no.1
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• pp.87-96
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• 2014

In the present study, a yeast species isolated from CETP, Vellore, Tamilnadu was identified as Cryptococcus sp. VITGBN2 based on molecular techniques and was found to be a potent producer of acidic diacetate sophorolipid in mineral salt media containing vegetable oil as additional carbon source. The chemical structure of the purified biosurfactant was identified as acidic diacetate sophorolipid through GC-MS analysis. This sophorolipid was used as a stabilizer for synthesis of zinc oxide nanoparticles (ZON). The formation of biofunctionalized ZON was characterized using UV-visible spectroscopy, XRD, scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy. The antimicrobial activities of naked ZON and sophorolipid functionalized ZON were tested based on the diameter of inhibition zone in agar well diffusion assay, microbial growth rate determination, protein leakage analysis, and lactate dehydrogenase assay. Bacterial pathogen Salmonella enterica and fungal pathogen Candida albicans showed more sensitivity to sophorolipid biofunctionalized ZON compared with naked ZON. Among the two pathogens, S. enterica showed higher sensitivity towards sophorolipid biofunctionalized ZON. SEM analysis showed that cell damage occurred through cell elongation in the case of S. enterica, whereas cell rupture was found to occur predominantly in the case of C. albicans. This is the first report on the dual role of yeast-mediated sophorolipid used as a biostabilizer for ZON synthesis as well as a novel functionalizing agent showing antimicrobial property.

• Journal of Microbiology and Biotechnology
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• v.27 no.2
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• pp.342-349
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• 2017

Polylactic acid (PLA) has been highlighted as an alternative renewable polymer for the replacement of petroleum-based plastic materials, and is considered to be biodegradable. On the other hand, the biodegradation of PLA by terminal degraders, such as microorganisms, requires a lengthy period in the natural environment, and its mechanism is not completely understood. PLA biodegradation studies have been conducted using mainly undefined mixed cultures, but only a few bacterial strains have been isolated and examined. For further characterization of PLA biodegradation, in this study, the PLA-degrading bacteria from digester sludge were isolated and identified using a polymer film-based screening method. The enrichment of sludge on PLA granules was conducted with the serial transference of a subculture into fresh media for 40 days, and the attached biofilm was inoculated on a PLA film on an agar plate. 3D optical microscopy showed that the isolates physically degraded the PLA film due to bacterial degradation. 16S rRNA gene sequencing identified the microbial colonies to be Pseudomonas sp. MYK1 and Bacillus sp. MYK2. The two isolates exhibited significantly higher specific gas production rates from PLA biodegradation compared with that of the initial sludge inoculum.

• KSBB Journal
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• v.19 no.2
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• pp.148-153
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• 2004

Lipase catalyzes the hydrolysis of glycerides into fatty acids and glycerol. The study of microbial lipases has been stimulated in resent years. It is due to the potential uses of lipases in esterification of oils to glycerol, alcohols and carbohydrates. Development of lipase producing yeast has been focused concerning to the utilization of yeast culture for animal feed. In this study, yeast like cells was isolated from a waste oil and sludge. A strain having higher lipase activity was selected by random mutagenesis using UV-radiation. The optimal cultivation conditions in submerged culture were examined in terms of lipase production. 2.0％ of high fructose syrup, 1,0％ of CSL, and 1.0％ of olive oil were selected as the nutritional media for the production of lipase. The maximum lipase activity of 1.12 U/ml and viable cell number of 8.8${\times}$10$\^$7/ cells/mL were obtained at 27$^{\circ}C$ with an initial pH of 5.0.

• Journal of Microbiology and Biotechnology
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• v.21 no.11
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• pp.1174-1178
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• 2011

Exopolysaccharides (EPSs) are microbial polysaccharides that are released outside of the bacterial cell wall. There have been few studies on EPS-producing lactic acid bacteria that can enhance macrophage activity and the underlying signaling mechanism for cytokine expression. In the current study, EPS-overproducing Lactobacillus (L.) paracasei KB28 was isolated from kimchi and cultivated in conditioned media containing glucose, sucrose, and lactose. The whole bacterial cells were obtained with their EPS being attached, and the cytokine-inducing activities of these cells were investigated. Gas chromatography analysis showed the presence of glucose, galactose, mannose, xylose, arabinose, and rhamnose in EPS composition. EPS-producing L. paracasei KB28 induced the expression of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, and IL-12 in mouse macrophages. This strain also caused the degradation of $I{\kappa}B{\alpha}$ and phosphorylation of the major MAPKs: Jun N-terminal kinase (JNK), p38, and extracellular signal-regulated kinase (ERK)1/2. The use of pharmacological inhibitors showed that different signaling pathways were involved in the induction of TNF-${\alpha}$, IL-6 and IL-12 by L. paracasei KB28. Our results provide information for a better understanding of the molecular mechanisms of the immunomodulatory effect of food-derived EPS-producing lactic acid bacteria.

• Korean Journal of Microbiology
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• v.36 no.3
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• pp.209-215
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• 2000

The purpose of U7is work was to iilvestigate the degradation of s-h~azine hel-hicidcs, ahilzine and simazine by TNT-degrader under several relevaut physicochemical environ~nental parameters. TNT-degrader showed effective degradability of atrazine and snnazine as well. Both atrazme (GO 1i1~11) and simazine ( 4 5 rng//) were completely degraded within 30 hrs and 4 days of incubation, respectively. As d ~ e concentrations of atrazine and sunazine increased in the media, the degradation ofthose compounds were delayed. Additional caubans were essential to degrade atrazine and simazule, and no degradation was achieved in the absence of additional carbons. The effect of supplemented nitrogens on the degradation of atrazine and sunazine was evalualed. Addition of a suppleinented nitrogen in he growth medium containing ah-azine or siinazine showed partial degr-adation olihose herbicides duriug the incubation period. However, complete degradation of atrazine and simazu~e was examined ul the absence or any supplemented nitrogens. Addltion of yeast extract in this study was inhibilory to atrazine aud siinazine degradations, respectively. TNT-degrader was a small Gram-negative cocco-bacillus. Physiological analysis using BIOLOG sysleln revealed that this strain was Ste~~ol~~opl~orno~~ns rrialtophilia.

• Microbiology and Biotechnology Letters
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• v.46 no.4
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• pp.390-394
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• 2018

For the coexpression of endoxylanase and endoglucanase genes in yeast Saccharomyces cerevisiae, the genes were separately inserted downstream of the yeast ADH1 promoters, resulting the plasmid pAGX3 (9.83 kb). In the batch culture on YPD medium of the yeast transformant, S. cerevisiae SEY2102/pAGX3, the total activities of the enzymes reached about 7.91 units/ml for endoxylanase and 0.43 units/ml for endoglucanase. In the fed-batch culture with intermittent feeding of yeast extract and glucose, the total activities of 24.9 units/ml for endoxylanase and 0.84 units/ml for endoglucanase were produced which were about 3.1-fold and 2.0-fold increased levels, respectively, compared to those of the batch culture. Most of endoxylanase and endoglucanase activities were found in the extracellular media. This recombinant yeast could be useful for the development of simultaneous saccharification bioprocess of the cellulose and xylan mixture.

• The Journal of Advanced Prosthodontics
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• v.11 no.2
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• pp.105-111
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• 2019

PURPOSE. Limited data is available regarding the differences for possible microleakage problems and fitting accuracy of zirconia versus titanium abutments with various connection designs. The purpose of this in vitro study was to investigate the effect of connection design and abutment material on the sealing capability and fitting accuracy of abutments. MATERIALS AND METHODS. A total of 42 abutments with different connection designs [internal conical (IC), internal tri-channel (IT), and external hexagonal (EH)] and abutment materials [titanium (Ti) and zirconia (Zr)] were evaluated. The inner parts of implants were inoculated with $0.7{\mu}L$ of polymicrobial culture (P. gingivalis, T. forsythia, T. denticola and F. nucleatum) and connected with their respective abutments under sterile conditions. The penetration of bacteria into the surrounding media was assessed by the visual evaluation of turbidity at each time point and the number of colony forming units (CFUs) was counted. The marginal gap at the implant- abutment interface (IAI) was measured by scanning electron microscope. The data sets were statistically analyzed using Kruskal-Wallis followed by Mann-Whitney U tests with the Bonferroni-Holm correction (${\alpha}=.05$). RESULTS. Statistically significant difference was found among the groups based on the results of leaked colonies (P<.05). The EH-Ti group characterized by an external hexagonal connection were less resistant to bacterial leakage than the groups EH-Zr, IT-Zr, IT-Ti, IC-Zr, and IC-Ti (P<.05). The marginal misfit (in ${\mu}m$) of the groups were in the range of 2.7-4.0 (IC-Zr), 1.8-5.3 (IC-Ti), 6.5-17.1 (IT-Zr), 5.4-12.0 (IT-Ti), 16.8-22.7 (EH-Zr), and 10.3-15.4 (EH-Ti). CONCLUSION. The sealing capability and marginal fit of abutments were affected by the type of abutment material and connection design.

• Journal of Microbiology and Biotechnology
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• v.32 no.3
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• pp.378-386
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• 2022

Scenedesmus obliquus ABC-009 is a microalgal strain that accumulates large amounts of lutein, particularly when subjected to growth-limiting conditions. Here, the performance of this strain was evaluated for the simultaneous production of lutein and biofuels under three different modes of cultivation - photoautotrophic mode using BG-11 medium with air or 2% CO₂ and heterotrophic mode using YM medium. While it was found that the highest fatty acid methyl ester (FAME) level and lutein content per biomass (%) were achieved in BG-11 medium with CO₂ and air, respectively, heterotrophic cultivation resulted in much higher biomass productivity. While the cell concentrations of the cultures grown under BG-11 and CO₂ were largely similar to those grown in YM medium, the disparity in the biomass yield was largely attributed to the larger cell volume in heterotrophically cultivated cells. Post-cultivation light treatment was found to further enhance the biomass productivity in all three cases and lutein content in heterotrophic conditions. Consequently, the maximum biomass (757.14 ± 20.20 mg/l/d), FAME (92.78 ± 0.08 mg/l/d), and lutein (1.006 ± 0.23 mg/l/d) productivities were obtained under heterotrophic cultivation. Next, large-scale lutein production using microalgae was demonstrated using a 1-ton open raceway pond cultivation system and a low-cost fertilizer (Eco-Sol). The overall biomass yields were similar in both media, while slightly higher lutein content was obtained using the fertilizer owing to the higher nitrogen content.

• 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.