• Journal of Animal Environmental Science
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• v.19 no.2
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• pp.155-162
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• 2013

The objective of this study is to investigate the effects of Cynanchum wilfordii (CW) on cell viability, anti-oxidant activity, volatile fatty acid (VFA) production and methane gas production. Collected rumen fluid incubated with CW powder (1% w/v) for 12 and 24 hours were analyzed for pH, VFAs and methane. Alamar blue assay showed no significant difference on the viability of 3T3-L1 and C2C12 cells treated with CW for 24 hours. TBARS data showed a dose dependent increase on the antioxidant activity of CW. VFAs increased in the CW-treated groups compared to the control group. In addition, propionate increased more than other VFAs by the treatment with CW. There was a significant decrease in methane gas production in batch culture treated with CW in 12hrs. In conclusion, it was suggested that Cynanchum wilfordii could manipulate rumen fermentation considered by increasing VFA production and inhibition of methanogenesis.

• Journal of Microbiology and Biotechnology
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• v.15 no.3
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• pp.461-465
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• 2005

Characteristics of biological $CO_2$ fixation by Chlorella sp. HA-1 were investigated in a semi-continuous and series reactor system using an internally illuminated photobioreactor to overcome shortcomings of physicochemical technologies such as adsorption and membrane separation. High $CO_2$ fixation rate was achieved in the semi-continuous reactor system, in which the dilution ratios of the culture medium were controlled. The average $CO_2$ fixation rate was maintained almost constantly when the dilution ratio increased by 0.1 increment from the initial value of 0.5. The total removal efficiency of $CO_2$ was enhanced by employing a series reactor system. The average $CO_2$ fixation rate increased until 4.013 g $CO_2\;day^{-1}$ in a series operation of four reactors, compared to 0.986 g $CO_2\;day^{-1}$ in a batch operation mode. The total $CO_2$ fixation rate was proportional to the number of reactors used in the series reactor system. In the series reactor system of semi-continuous operation, a large amount of $CO_2$ was removed continuously for 30 days. These results showed that the present reactor systems are efficient and economically feasible for a biological $CO_2$ fixation.

• Applied Chemistry for Engineering
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• v.25 no.4
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• pp.386-391
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• 2014

Atmospheric-pressure nonthermal corona discharge plasma was applied to the sterilization of biologically contaminated scoria powder. Escherichia coli (E. coli) culture solution was uniformly sprayed throughout the scoria powder for artificial inoculation, which was well mixed to ensure uniformity of the batch. The effect of the key parameters such as discharge power, treatment time, type of gas and electrode distance on the sterilization efficiency was examined and discussed. The experimental results revealed that the plasma treatment was very effective for the sterilization of scoria powder; 5-min treatment at 15 W could sterilize more than 99.9% of E. coli inoculated into the scoria powder. Increasing the discharge power, treatment time or applied voltage led to an improvement in the sterilization efficiency. The effect of type of gas on the sterilization efficiency was in order of oxygen, synthetic air (20% oxygen) and nitrogen from high to low. The inactivation of E. coli under the influence of corona discharge plasma can be explained by cell membrane erosion or etching resulting from UV and reactive oxidizing species (oxygen radical, OH radical, ozone, etc.), and the destruction of E. coli cell membrane by the physical action of numerous corona streamers.

• Applied Biological Chemistry
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• v.34 no.3
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• pp.273-278
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• 1991

Methanol assimilating bacterium, Pseudomonas sp. ILS-003 was used to investigate the optimum conditions for the production of $poly-{\beta}-hydroxybutyric$ acid from methanol. For PHB production, the optimum initial pH was 6.4 and the optimum temperature was $30^{\circ}C$. Also the optimum methanol concentration was found to be 1.0%(v/v). In the PHB production, $(NH_4)_2SO_4$ was the most effective nitrogen source and the optimum concentration was 0.8 g/l, which was eqivalent to 17.4 in C/N ratio. Also, deficiency of the 2 valence metal ions in the medium had stimulating effect on PHB accumulation. Under the optimum substrate concentration, successive feeding of 0.25%(v/v) methanol was the most effective on PHB production. Under the optimum conditions, 1.94 g/l of PHB and 2.78 g/l of dry biomass were produced in 96 hours, and the yield was 69.8%(w/w).

• Journal of Microbiology and Biotechnology
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• v.18 no.11
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• pp.1773-1784
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• 2008

Effects of various industrially important carbon sources (glucose, sucrose, xylose, gluconate, and glycerol) on shikimic acid (SA) biosynthesis in Escherichia coli were investigated to gain new insight into the metabolic capability for overproducing SA. At the outset, constraints-based flux analysis using the genome-scale in silico model of E. coli was conducted to quantify the theoretical maximum SA yield. The corresponding flux distributions fueled by different carbon sources under investigation were compared with respect to theoretical yield and energy utilization, thereby identifying the indispensable pathways for achieving optimal SA production on each carbon source. Subsequently, a shikimate-kinase-deficient E. coli mutant was developed by blocking the aromatic amino acid pathway, and the production of SA on various carbon sources was experimentally examined during 51 batch culture. As a result, the highest production rate, 1.92 mmol SA/h, was obtained when glucose was utilized as a carbon source, whereas the efficient SA production from glycerol was obtained with the highest yield, 0.21 mol SA formed per mol carbon atom of carbon source consumed. The current strain can be further improved to satisfy the theoretically achievable SA production that was predicted by in silico analysis.

• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1516-1524
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• 2014

Flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) can utilize a variety of external electron acceptors and also has stricter substrate specificity than any other glucose oxidoreductases, which makes it the ideal diagnostic enzyme in the field of glucose biosensors. A gene coding for a hypothetical protein, similar to glucose oxidase and derived from Aspergillus terreus NIH2624, was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 260,000 U/l in the culture supernatant after fed-batch cultivation for 84 h. After a three-step purification protocol that included isopropanol precipitation, affinity chromatography, and a second isopropanol precipitation, recombinant FAD-GDH was purified with a recovery of 65%. This is the first time that isopropanol precipitation has been used to concentrate a fermentation supernatant and exchange buffers after affinity chromatography purification. The purified FAD-GDH exhibited a broad and diffuse band between 83 and 150 kDa. The recombinant FAD-GDH was stable across a wide pH range (3.5 to 9.0) with maximum activity at pH 7.5 and $55^{\circ}C$. In addition, it displayed very high thermal stability, with a half-life of 82 min at $60^{\circ}C$. These characteristics indicate that FAD-GDH will be useful in the field of glucose biosensors.

• Journal of Microbiology and Biotechnology
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• v.23 no.3
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• pp.351-356
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• 2013

A high ${\beta}$-glucosidase (BGL)-producing strain, Stereum hirsutum, was identified and isolated and showed a maximum BGL activity (10.4 U/ml) when cultured with Avicel and tryptone as the carbon and nitrogen sources, respectively. In comparison with other BGLs, BGL obtained from S. hirsutum showed a higher level of activity to cellobiose ($V_{max}$ = 172 U/mg, and $k_{cat}$ = 281/s). Under the optimum conditions (600 rpm, $30^{\circ}C$, and pH 6.0), the maximum BGL activity of 10.4 U/ml with the overall productivity of 74.5 U/l/h was observed. BGL production was scaled up from a laboratory scale (7-L fermenter) to a pilot scale (70-L fermenter). When S. hirsutum was cultured in fed-batch culture with rice straw as the carbon source in a 70-L fermenter, a comparable productivity of 78.6 U/l/h was obtained. Furthermore, S. hirsutum showed high levels of activity of other lignocellulases (cellobiohydrolase, endoglucanase, xylanase, and laccase) that are involved in the saccharification of biomasses. Application of S. hirsutum lignocellulases in the hydrolysis of Pinus densiflora and Catalpa ovata showed saccharification yields of 49.7% and 43.0%, respectively, which were higher than the yield obtained using commercial enzymes.

• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1281-1287
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• 2017

Bone morphogenetic protein-4 (BMP-4) is considered to have therapeutic potential for various diseases, including cancers; however, the high expression of biologically active recombinant human BMP-4 (rhBMP-4) needed for its manufacture for therapeutic purposes has yet to be established. In the current study, we established a recombinant Chinese hamster ovary (rCHO) cell line overexpressing rhBMP-4 as well as a production process using 7.5-l bioreactor (5 L working volume). The expression of the mature rhBMP-4 was significantly enhanced by recombinant furin expression. The combination of a chemically defined medium and a nutrient supplement solution for high expression of rhBMP-4 was selected and used for bioreactor cultures. The 11-day fed-batch cultures of the established rhBMP-4-expressing rCHO cells in the 7.5-L bioreactor produced approximately 32 mg/l of rhBMP-4. The mature rhBMP-4 was purified to homogeneity from the culture supernatant using a two-step chromatographic procedure, resulting in a recovery rate of approximately 55% and a protein purity greater than 95%. The N-terminal amino acid sequences and N-linked glycosylation of the purified rhBMP-4 were confirmed by N-terminal sequencing and de-N-glycosylation analysis, respectively. The mature purified rhBMP-4 has been proved to be functionally active, with an effective dose concentration of $EC_{50}$ of 2.93 ng/ml.

• Journal of the korean academy of Pediatric Dentistry
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• v.33 no.1
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• pp.77-84
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• 2006

Since the long-term exposure of mutans streptococci to xylitol is known to select for xylitol-resistant $(X^R)$ natural mutants, the occurrence and survival of such $(X^R)$ strains were performed in batch culture methods. The aim of the study was to compare the differentiation and quantification of mRNA expression of the gtf genes of $X^R\;and\;X^S$ mutans streptococci. Using a real-time reverse-transcription polymerase chain reaction, the expression of each gtf was determined. In $X^R$ strains, the relative levels of transcription of gtfB and gtfC were decreased while that of gtfD was increased, suggesting the presence of independent promoters. It also suggested that mutation related to production of glucosyltransferase occurred under the exposure of xylitol could explain the caries-preventive mechanisms of xylitol.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.2
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• pp.144-149
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• 2001

For the production and purification of a single chain human insulin precursor, four types of fusion peptides $\beta$-galactosidase (LacZ), maltose binding protein (MBP), glutathione-S-transferase (GST), and (His)(sub)6-tagged sequence (HTS) were investigated. Recombinant E. coli harboring hybrid genes was cultivated at 37$\^{C}$ for 1h, and gene induction occurred when 0.2mM of isopropyl-D-thiogalactoside (IPTG) was added to the culture broth, except for E. coli BL21 (DE3) pLysS harboring a pET-BA cultivation with 1.0mM IPTG, followed by a longer than 4h batch fermentation respectively. DEAE-Sphacel and Sephadex G-200 gel filtration chromatography, amylose affinity chromatography, glutathione-sepharose 4B affinity chromatography, and a nickel chelating affinity chromatography system as a kind of immobilized metal ion affinity chromatography (IMAC) were all employed for the purification of a single chain human insulin precursor. The recovery yields of the HTS-fused, GST-fused, MBP-fused, and LacZ-fused single chain human insulin precursors resulted in 47%, 20%, 20%, and 18% as the total protein amounts respectively. These results show that a higher recovery yield of the finally purified recombinant peptides was achieved when affinity column chromatography was employed and when the fused peptide had a smaller molecular weight. In addition the pET expression system gave the highest productivity of a fused insulin precursor due to a two-step regulation of the gene expression, and the HTS-fused system provided the highest recovery of a fused insulin precursor based on a simple and specific separation using the IMAC technique.