• Journal of Life Science
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• v.17 no.6 s.86
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• pp.847-858
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• 2007

The endoplasmic reticulum (ER) is an important intracellular organelle for folding and maturation of newly synthesized transmembrane and secretory proteins. The ER provides stringent quality control systems to ensure that only correctly folded proteins exit the ER and unfolded or misfolded proteins are retained and ultimately degraded. The ER has evolved stress response both signaling pathways the unfolded protein response (UPR) to cope with the accmulation of unfolded or misfolded proteins and ER overload response (EOR). Accumulating evidence suggests that, in addition to responsibility for protein processing, ER is also an important signaling compartment and a sensor of cellular stress. In this respect, production of bio-functional recombinant-proteins requires efficient functioning of the ER secretory pathway in host cells. This review briefly summarizes our understanding of the ER signaling developed in the recent years to help of the secretion capacities of recombinant cells.

• Journal of Environmental Health Sciences
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• v.35 no.3
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• pp.214-219
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• 2009

This study examined the removal efficiency of the nitrogen and phosphorus in order to compensate for the combined process of ATAD(Autothermal Thermophilic Aaerobic Digestion) and EGSB(Expended Granular Sludge Bed), which are treatment methods for livestock wastewater, by introducing SBR(Sequencing Batch Reactor) as post-treatment process. The analysis on the treatment efficiency of each operation mode showed that, in the case of T-N, the treatment efficiency were 67.1% and 74.2% for Run-1 and Run-2, respectively, and in the case of T-P, the values were 71.2 and 74.1, respectively, which are indicating that the treatment efficacy is higher in the condition of Run-1, in which the time period of Anoxic and Aerobic segments were increased. In addition, the result of analyzing removal characteristics of nitrogen and phosphorus by Influx load showed that removal efficiency of nitrogen was better in the case of high influx load than in the case of low influx load. Regardless of Influx load, phosphorus showed constant influx concentration, so that removal efficiency of phosphorus was influenced littler by Influx load than that of nitrogen. This study also fed methanol as an external carbon source and performed experiment to induce denitrification under anoxic condition by using nitrate among nitrogen compounds of SBR reactor, and the results showed that intermittent feeding was more effective in Nitrogen Removal than composite feeding.

• KSBB Journal
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• v.10 no.3
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• pp.343-348
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• 1995

Human lipocorin-I(LCI) is a calcium ion-dependent and phospholipid-binding protein which exhibits an anti-inflammatory activity by inhibiting phospholipase A2 activity. In this study, the LCI gene containing its own terminator region was joined to GAL10 promoter-ppL (prepro-leader sequence of mating factor a). An ATG start codon of LCI gene was placed at downstream with KR endoprotease recognition site(Lys-Arg) of ppL. Recombinant S. cerevisiae harboring the LCI expression/secretion vector, pYGLPT5, was aerobicall grown on a liquid YPDG medium al $30^{\circ}C$ for 72hys. The whole cell and culture supernatant were separated after centrifugation, and the expressed LCI was analyzed by SDS-PAGE and western blotting methods. A majority fraction of the expressed LCI was found to be accumulated in the intracellular fraction, resulting in very low secretion efficiency of about 7.4%. About $500mg/\ell$ of LCI was extracellularly produced by the fed-batch culture employing the controlledfeeding of glucose and galactose. The secreted LCI was purified by ultrafiltration and hydroxylapatite column chromatography, and a purity of more than 99% was obtained.

• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.123-127
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• 2004

Pseudomonas sp. MBEL21 was newly isolated from soil samples and found to accumulate medium-chain-length Polyhydroxyalkanoates(MCL-PHAs) using oleic acid as a sole carbon source. Among the various nutrient limiting conditions examined, including nitrogen, sulfur and phosphorus, only phosphorus limitation supported the accumulation of MCL-PHAs up to 15 wt％ of dry cell weight in flask cultures. MCL-PHAs produced by Pseudomonas sp. MBEL21 was mainly composed of 3-hydroxy-5-cis-tetradecenoate. Fed-batch culture of Pseudomonas sp. MBEL21 by novel feeding strategies based on cell growth charcteristics was carried out under phosphorus limitation using oleic acid as a sole carbon source. The final cell concentration and PHA content of 82 g/L and 28 wt％, respectively, were obtained. Furthermore, PHA consisted of MCL-hydroxyalkanoates and 3-hydroxybutyrate could be produced using olive oil as a sole carbon source.

• Korean Journal of Food Science and Technology
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• v.51 no.4
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• pp.343-347
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• 2019

This study was performed to develop an economical culture method of lactic acid bacteria (LAB) for kimchi fermentation. Leuconostoc mesenteroides WiKim32 was grown to $1{\times}10^9CFU/mL$ and maintained at 8.88 log CFU/mL for four days by culturing in kimchi cabbage juice (KCJ) without supplements and sterilization. Leuconostoc mesenteroides WiKim32 was cultured in 100 mL of KCJ by inoculating with 0.1% starter culture and adding 100 mL of KCJ everyday by adjusting pH to 5.5 using 1 M NaOH at $20^{\circ}C$ for four days. KCJ was prepared by extraction of kimchi cabbage leaves after washing them with citric acid and ethanol. Adjusting pH over 6.0 was favorable for the growth of LAB in the initial stage, but LAB growth was retarded in the later stage. In contrast, adjusting pH below 5.0 was not beneficial for the growth of LAB; therefore, pH was adjusted to 5.5.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.6
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• pp.776-782
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• 2019

Objective: Fasting may lead to changes in the microbiota and activity in the rumen. In the present study, the effects of fasting on rumen microbiota and the impact of fasting on in vitro rumen fermentation were evaluated using molecular culture-independent methods. Methods: Three ruminally cannulated Holstein steers were fed rice straw and concentrates. The ruminal fluids were obtained from the same steers 2 h after the morning feeding (control) and 24 h after fasting (fasting). The ruminal fluid was filtrated through four layers of muslin, collected for a culture-independent microbial analysis, and used to determine the in vitro rumen fermentation characteristics. Total DNA was extracted from both control and fasting ruminal fluids. The rumen microbiota was assessed using denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction. Microbial activity was evaluated in control and fasting steers at various intervals using in vitro batch culture with rice straw and concentrate at a ratio of 60:40. Results: Fasting for 24 h slightly affected the microbiota structure in the rumen as determined by DGGE. Additionally, several microorganisms, including Anaerovibrio lipolytica, Eubacterium ruminantium, Prevotella albensis, Prevotella ruminicola, and Ruminobacter amylophilus, decreased in number after fasting. In addition, using the ruminal fluid as the inoculum after 24 h of fasting, the fermentation characteristics differed from those obtained using non-fasted ruminal fluid. Compared with the control, the fasting showed higher total gas production, ammonia, and microbial protein production (p<0.05). No significant differences, however, was observed in pH and dry matter digestibility. Conclusion: When in vitro techniques are used to evaluate feed, the use of the ruminal fluid from fasted animals should be used with caution.

• Journal of Environmental Science International
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• v.31 no.2
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• pp.141-148
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• 2022

In this study, the inhibition of ammonia on anaerobic digestion of butyric acid was evaluated and the potential alleviating effects of such ammonia inhibition by the addition of magnetite particles were investigated. Independent anaerobic batch tests fed with butyric acid as a sole organic source were conducted in twenty 60-mL glass bottles with 10 different treatment conditions, comprising ammonia: 0.5, 2.0, 4.0, 6.0, and 7.0 g total ammonia nitrogen (TAN)/L and magnetite particles: 0 mM and 20 mM. The increase in ammonia concentration did not cause significant inhibition on methane yield; however, a significant inhibition on lag time and specific methane production rate was observed. The IC₅₀ in the control treatments (without magnetite addition) was estimated as 6.2654 g TAN/L. A similar inhibition trend was observed in magnetite-added treatments; however, the inhibition effect by ammonia was significantly alleviated in lag time and specific methane production rate when compared to those in the control treatments. The lag time was shortened by 1.6-46.3%, specific methane production rate was improved by 6.0-69.0%. In the magnetite-added treatments, IC₅₀ was estimated as 8.5361 g TAN/L. This study successfully demonstrated the potential of magnetite particles as an enhancer in anaerobic digestion of butyric acid under conditions of ammonia stress.

• Korean Chemical Engineering Research
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• v.47 no.6
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• pp.768-774
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• 2009

To improve the productivity of 1,3-propanediol(1,3-PD) with K. pneumoniae DSM4799 using pure glycerol and crude glycerol derived from the biodiesel process, optimizing fermentation conditions was performed by changing environmental factors such as anaerobic/aerobic condition, temperature, glycerol concentration, and pH. When anaerobic conditions were maintained, there was an improved 1,3-PD production compared with that from aerobic/anaerobic 2-stage fermentation. From the results with temperature $26{\sim}37^{\circ}C$, the higher 1,3-PD production yield was observed at $30{\sim}33^{\circ}C$. For an initial glycerol concentration higher than 60 g/L, cell growth and 1,3-PD production were inhibited. When crude glycerol was used, the initial 1,3-PD production appeared to be inhibited. After 48 hr of incubation, however, 1,3-PD production with crude glycerol was even higher than that with pure glycerol, demonstrating the feasibility of 1,3-PD production using crude glycerol as a substrate. Fed-batch fermentation was applied for the high concentration of 1,3-PD without substrate inhibition. By regulating pH at 7 during the fed-batch with glycerol lower than 40 g/L, the yield of 1,3-PD was 25% higher than that without pH regulation(0.56 g/g vs. 0.45 g/g). In conclusion, based on our results, anaerobic conditions, temperature at $30^{\circ}C$, pure or crude glycerol lower than 40 g/L, and pH regulation at 7 were the optimized conditions for 1,3-PD production using K. pneumoniae DSM4799, making it more feasible to produce 1,3-PD at higher concentration and a lower price.

• Journal of Korean Society of Water and Wastewater
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• v.27 no.6
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• pp.689-701
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• 2013

Bacterial growth and corresponding consumption of carbon and phosphorus were examined in which tap water samples containing a very low concentration of free chlorine were supplemented with organic carbon and/or phosphorus. The experiments were performed in a fed-batch mode under a controlled temperature of $20^{\circ}C$. In the phosphorus alone-added water, there was no significant increase in bacterial numbers measured as heterotrophic plate count (HPC) in the bulk water. However, bacterial growth was stimulated by the addition of carbon (e.g., bulk HPC levels increased to $10^3CFU/mL$) and further stimulated by the combined addition of carbon and phosphorus (e.g., bulk HPC to $10^5CFU/mL$). The same effects were observed in biofilm HPC and biomass formed on polyethylene (PE) slide surfaces. In the water where organic carbon and phosphorus were added together, the highest biofilm HPC and biomass (measured as extracellular polymeric substance components) densities were observed which were $7.6{\times}10^5CFU/cm^2$ and $5.3{\mu}g/cm^2$, respectively. In addition to the bacterial growth, additions of organic carbon and/or phosphorus resulted in different bacterial carbon-to-phosphorus (C/P) consumption ratios. Compared to a typical bacterial C/P consumption ratio of 100:1, a higher C/P ratio (590:1) occurred in the carbon alone-added water, while a lower ratio (40:1) in phosphorus alone-added water. Comparative value (80:1) of C/P ratio was also observed in the water where organic carbon and phosphorus were added together. At the given experimental conditions, bacterial growth was deemed to be more sensitive to microbially available organic carbon than phosphorus. The effect of phosphorus addition, which resulted in a lower C/P consumption ratio, seemed to be tightly associated with the presence of microbially available organic carbon. These results suggested that the control of extrinsic carbon influx seemed to be more important to minimize bacterial regrowth in drinking water system, since even low content of phosphorus naturally occurring in drinking water was enough to allow a bacterial growth.

• KSBB Journal
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• v.21 no.3
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• pp.204-211
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• 2006

For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.