• Journal of Microbiology and Biotechnology
• /
• 제30권9호
• /
• pp.1430-1435
• /
• 2020

Bacterial cellulose (BC) has outstanding physical and chemical properties, including high crystallinity, moisture retention, and tensile strength. Currently, the major producer of BC is Komagataeibacter xylinus. However, due to limited tools of expression, this host is difficult to engineer metabolically to improve BC productivity. In this study, a regulated expression system for K. xylinus with synthetic ribosome binding site (RBS) was developed and used to engineer a BC biosynthesis pathway. A synthetic RBS library was constructed using green fluorescent protein (GFP) as a reporter, and three synthetic RBSs (R4, R15, and R6) with different strengths were successfully isolated by fluorescence-activated cell sorting (FACS). Using synthetic RBS, we optimized the expression of three homologous genes responsible for BC production, pgm, galU, and ndp, and thereby greatly increased it under both static and shaking culture conditions. The final titer of BC under static and shaking conditions was 5.28 and 3.67 g/l, respectively. Our findings demonstrate that reinforced metabolic flux towards BC through quantitative gene expression represents a practical strategy for the improvement of BC productivity.

• 한국전기전자재료학회:학술대회논문집
• /
• 한국전기전자재료학회 2006년도 하계학술대회 논문집 Vol.7
• /
• pp.378-379
• /
• 2006

The carbon thin film was developed by the CVD method using the carbon source of toluene with the stream of argon gas at $800{\sim}1100^{\circ}C$ for 1 hour. Developed carbon thin films have the material loading of 0.27 mg($800^{\circ}C$), 0.80 mg($900^{\circ}C$), 2.3 mg($1000^{\circ}C$), and 2.9 mg($1100^{\circ}C$) for the disk of 15 mm diameter on single side. The characteristics of carbon thin film as the anode of thin film battery were evaluated using Li|C coin cell. Li|C($1100^{\circ}C$) coin cell has the first specific discharge and charge capacity of 953 mAh/g and 374 mAh/g, respectively, resulting the first Ah efficiency of 39.3 %. Capacity retention of the 5th cycle was 93.2 % indicating good cycleability. The carbon thin film prepared by CVD shows good specific capacity and cycleability, but low Ah efficiency.

• Biomolecules & Therapeutics
• /
• 제30권6호
• /
• pp.546-552
• /
• 2022

Epidermal cell adhesion molecule (EpCAM) is a tumor-associated antigen (TAA), which has been considered as a cancer vaccine candidate. The EpCAM protein fused to the fragment crystallizable region of immunoglobulin G (IgG) tagged with KDEL endoplasmic reticulum (ER) retention signal (EpCAM-FcK) has been successfully expressed in transgenic tobacco (Nicotiana tabacum cv. Xanthi) and purified from the plant leaf. In this study, we investigated the ability of the plant-derived EpCAM-FcK (EpCAM-FcK^P) to elicit an immune response in vivo. The animal group injected with the EpCAM-FcK^P showed a higher differentiated germinal center (GC) B cell population (~9%) compared with the animal group injected with the recombinant rhEpCAM-Fc chimera (EpCAM-Fc^M). The animal group injected with EpCAM-FcK^P (~42%) had more differentiated T follicular helper cells (Tfh) than the animal group injected with EpCAM-Fc^M (~7%). This study demonstrated that the plant-derived EpCAM-FcK fusion antigenic protein induced a humoral immune response in mice.

• 멤브레인
• /
• 제33권6호
• /
• pp.416-426
• /
• 2023

본 연구에서는 산화 방지 특성이 있는 가리워진 아민기를 함유한 산화 그래핀(hindered amine grafted graphene oxide, HA-GO)을 합성하여 이를 도입한 나피온(Nafion) 기반의 복합 막을 제조한 후 고분자 전해질 막 연료전지 시스템에 응용하였다. HA-GO는 4-아미노-2, 2, 6, 6-테트라메틸-4-피페리딘(4-amino-2, 2, 6, 6-tetramethyl piperidine)에 존재하는 아민기와 GO 표면에 존재하는 에폭시기의 개환 반응을 통해 제조하였으며, 합성된 HA-GO의 함량을 달리한 복합 막을 제조하여 순수 Nafion 막과 성능 특성을 비교하였다. HA-GO가 첨가된 복합 막은 Nafion 단일 막에 비해 기계적 물성, 화학적 안정성 및 수소이온 전도 특성이 향상되었다. 특히 HA-GO의 산화 방지 특성으로 인해 HA-GO가 첨가된 복합 막은 펜톤 평가(Fenton's test) 이후 수소이온 전도도의 유지 특성이 Nafion 단일 막에 비해 큰 폭으로 향상된 것을 확인할 수 있었다.

• Molecules and Cells
• /
• 제46권12호
• /
• pp.764-777
• /
• 2023

Recombinant immunotoxins (RITs) are fusion proteins consisting of a targeting domain linked to a toxin, offering a highly specific therapeutic strategy for cancer treatment. In this study, we engineered and characterized RITs aimed at mesothelin, a cell surface glycoprotein overexpressed in various malignancies. Through an extensive screening of a large nanobody library, four mesothelin-specific nanobodies were selected and genetically fused to a truncated Pseudomonas exotoxin (PE24B). Various optimizations, including the incorporation of furin cleavage sites, maltose-binding protein tags, and tobacco etch virus protease cleavage sites, were implemented to improve protein expression, solubility, and purification. The RITs were successfully overexpressed in Escherichia coli, achieving high solubility and purity post-purification. In vitro cytotoxicity assays on gastric carcinoma cell lines NCI-N87 and AGS revealed that Meso(Nb2)-PE24B demonstrated the highest cytotoxic efficacy, warranting further characterization. This RIT also displayed selective binding to human and monkey mesothelins but not to mouse mesothelin. The competitive binding assays between different RIT constructs revealed significant alterations in IC₅₀ values, emphasizing the importance of nanobody specificity. Finally, a modification in the endoplasmic reticulum retention signal at the C-terminus further augmented its cytotoxic activity. Our findings offer valuable insights into the design and optimization of RITs, showcasing the potential of Meso(Nb2)-PE24B as a promising therapeutic candidate for targeted cancer treatment.

• Journal of Microbiology and Biotechnology
• /
• 제34권4호
• /
• pp.969-977
• /
• 2024

Indigo is a valuable, natural blue dye that has been used for centuries in the textile industry. The large-scale commercial production of indigo relies on its extraction from plants and chemical synthesis. Studies are being conducted to develop methods for environment-friendly and sustainable production of indigo using genetically engineered microbes. Here, to enhance the yield of bioindigo from an E. coli whole-cell system containing tryptophanase (TnaA) and flavin-containing monooxygenase (FMO), we evaluated tryptophan transporters to improve the transport of aromatic compounds, such as indole and tryptophan, which are not easily soluble and passable through cell walls. Among the three transporters, Mtr, AroP, and TnaB, AroP enhanced indigo production the most. The combination of each transporter with AroP was also evaluated, and the combination of AroP and TnaB showed the best performance compared to the single transporters and two transporters. Bioindigo production was then optimized by examining the culture medium, temperature, isopropyl β-D-1-thiogalactopyranoside concentration, shaking speed (rpm), and pH. The novel strain containing aroP and tnaB plasmid with tnaA and FMO produced 8.77 mM (2.3 g/l) of bioindigo after 66 h of culture. The produced bioindigo was further recovered using a simple method and used as a watercolor dye, showing good mixing with other colors and color retention for a relatively long time. This study presents an effective strategy for enhancing indigo production using a combination of transporters.

• Journal of Electrochemical Science and Technology
• /
• 제15권3호
• /
• pp.365-372
• /
• 2024

As the energy density of lithium-ion batteries (LIBs) continues to increase, various separators are being developed to with the aim of improving the safety performance. Although poly(imide) (PI)-based separators are widely used, it is difficult to control their pore size and distribution, and this may further increase the risk associated. Herein, a melamine phosphonic acid (MP)-coated PI separator that can effectively control the pore structure of the substrate is suggested as a remedy. After the MP material is embedded into the PI separator with a simple one-step casting process, it effectively clogs the large pores of the PI separator, preventing the occurrence of internal short circuits during charging. It is anticipated that the MP material can also suppress rapid thermal runaway upon cycling due to its ability to reduce the internal temperature of the LIB cell caused by the desirable endothermic behavior around 300℃. According to experiments, the MP-coated PI separator not only decreases the thermal shrinkage rate better than commercial poly(ethylene) (PE) separators but also exhibits a desirable Gurley number (109.6 s/100 cc) and electrolyte uptake rate (240%), which is unique. The proposed separator is electrochemically stable in the range 0.0-5.0 V (vs. Li/Li⁺), which is the typical working potential of conventional electrode materials. In practice, the MP-coated PI separator exhibits stable cycling performance in a graphite-LiNi_0.83Co_0.10Mn_0.07O₂ full cell without an internal short circuit (retention: 90.3%).

• Bulletin of the Korean Chemical Society
• /
• 제32권5호
• /
• pp.1491-1494
• /
• 2011

Lithium vanadium phosphate, $Li_3V_2(PO_4)_3$ was prepared by a simple solid state route. It was found that making a fine powder of $Li_3V_2(PO_4)_3$ by the mechanical milling is very effective for increasing the insertion/extraction of lithium from $Li_3V_2(PO_4)_3$ structure. In charge/discharge test, the ball-milled $Li_3V_2(PO_4)_3$ sample exhibited a higher initial discharge capacity of 174 mAh/g in the voltage range of 3.0-4.8 V, compared with pure $Li_3V_2(PO_4)_3$ sample (152 mAh/g). Furthermore, the ball-milled $Li_3V_2(PO_4)_3$ presented not only higher cycle retention rate after 50 cycles, but also better rate capability compared with pure sample in the whole region (0.1-7 C).

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제9권1호
• /
• pp.47-51
• /
• 2004

Ceramide III was prepared by the cultivation of Saccharomyces cerevisiae. Ceramide III was partitioned from the cell extracts by solvent extraction and analyzed by Normal Phase High Performance Liquid Chromatography (NP-HPLC) using Evaporative Light Scattering Detector (ELSD). We experimentally determined the mobile phase composition to separate ceramide III with NP-HPLC. Three binary mobile phases of n-hexane/ethanol, n-hexane/lsoprophyl Alcohol(IPA) and n-hexane/n-butanol and one ternary mobile phase of n-hexane/IPA/methanol were demonstrated. For the binary mobile phase of n-hexane/ethanol, the first mobile phase composition, 95/5(v/v), was step-increased to 72/23(v/v) at 3 min. In the binary mobile phase, the retention time of ceramide III was 7.87min, while it was 4.11 min respectively in the ternary system, where the mobile phase composition of n-hexane/IPA/methanol, 85/7/8(v/v/v), was step-increased to 75/10/15(v/v/v) at 3 min. However, in the ternary mobile phase, the more peak area of ceramide III was observed.

• 동의생리병리학회지
• /
• 제18권6호
• /
• pp.1777-1783
• /
• 2004

This study was conducted to determine the effects Wonjiseokchangpo-san on brain damage in transient focal cerebral ischemia. Rats were used for testing in the following three models: Morris Water Maze, Eight-Arm Radial Maze, and Histochemistry. In the Morris Water Maze Model, the Wonjiseokchangpo-san group showed significant decrease in the 3rd and 6th training session compared with the ischemia group. A retention test, in the Morris Water Maze Model, was performed on the 7th day without the escape platform. The Wonjiseokchangpo-san group showed significant increase compared to the ischemia group. In the Eight-Arm radial Maze model, the Wonjiseokchangpo-san group showed significant decrease in the error rate compared to the ischemia group. In the density of hippocampal CA1 cell of the cresyl violet-stained section, the Wonjiseokahangpo-san group showed significant increase compared to the ischemia group. These results suggest that Wonjiseokchangpo-san may have a significant protective effect on brain damage and cognitive dysfunction in transient focal cerebral ischemia.