• Journal of Microbiology and Biotechnology
• /
• 제22권12호
• /
• pp.1613-1620
• /
• 2012

Bacterial wilt caused by Ralstonia solanacearum is a devastating disease of many economically important crops. Since there is no promising control strategy for bacterial wilt, phage therapy could be adopted using virulent phages. We used phage PE204 as a model lytic bacteriophage to investigate its biocontrol potential for bacterial wilt on tomato plants. The phage PE204 has a short-tailed icosahedral structure and double-stranded DNA genome similar to that of the members of Podoviridae. PE204 is stable under a wide range of temperature and pH, and is also stable in the presence of the surfactant Silwet L-77. An artificial soil microcosm (ASM) to study phage stability in soil was adopted to investigate phage viability under a controlled system. Whereas phage showed less stability under elevated temperature in the ASM, the presence of host bacteria helped to maintain a stable phage population. Simultaneous treatment of phage PE204 at $10^8$ PFU/ml with R. solanacearum on tomato rhizosphere completely inhibited bacterial wilt occurrence, and amendment of Silwet L-77 at 0.1% to the phage suspension did not impair the disease control activity of PE204. The biocontrol activities of phage PE204 application onto tomato rhizosphere before or after R. solanacearum inoculation were also investigated. Whereas pretreatment with the phage was not effective in the control of bacterial wilt, post-treatment of PE204 delayed bacterial wilt development. Our results suggested that appropriate application of lytic phages to the plant root system with a surfactant such as Silwet L-77 could be used to control the bacterial wilt of crops.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제7권1호
• /
• pp.43-51
• /
• 2002

The cloning and expression of $\beta-glucosidase$ II, encoded by the gene ${\beta}glu2$, from thermotolerant yeast Pichia etchellsii into Escherichia coli is described. Cloning of the 7.3 kb BamHI/SalI yeast insert containing ${\beta}glu2$ in pUC18, which allowed for reverse orientation of the insert, resulted in better enzyme expression. Transformation of this plasmid into E. coli JM109 resulted in accumulation of the enzyme in periplasmic space. At $50^{\circ}C$, the highest hydrolytic activity of 1686 IU/g protein was obtained on sophorose. Batch and fed-batch techniques were employed for enzyme production in a 14 L bioreactor. Exponential feeding rates were determined from mass balance equations and these were employed to control specific growth rate and in turn maximize cell growth and enzyme production. Media optimization coupled with this strategy resulted in increased enzyme units of 1.2 kU/L at a stabilized growth rate of $0.14\;h^{-l}$. Increased enzyme production in bioreactor was accompanied by formation of inclusion bodies.

• Journal of Microbiology and Biotechnology
• /
• 제29권6호
• /
• pp.923-932
• /
• 2019

Current strategies of strain improvement processes are mainly focused on enhancing the synthetic pathways of the products. However, excessive metabolic flux often creates metabolic imbalances, which lead to growth retardation and ultimately limit the yield of the product. To solve this problem, we applied a dynamic regulation strategy to produce $\text\tiny{L}$-phenylalanine ($\text\tiny{L}$-Phe) in Escherichia coli. First, we constructed a series of Phe-induced promoters that exhibited different strengths through modification of the promoter region of tyrP. Then, two engineered promoters were separately introduced into a Phe-producing strain xllp1 to dynamically control the expression level of one pathway enzyme AroK. Batch fermentation results of the strain xllp3 showed that the titer of Phe reached 61.3 g/l at 48 h, representing a titer of 1.36-fold of the strain xllp1 (45.0 g/l). Moreover, the $\text\tiny{L}$-Phe yields on glucose of xllp3 (0.22 g/g) were also greatly improved, with an increase of 1.22-fold in comparison with the xllp1 (0.18 g/g). In summary, we successfully improved the titer of Phe by using dynamic regulation of one key enzyme and this strategy can be applied for improving the performance of strains producing other aromatic amino acids and derived compounds.

• Journal of Microbiology and Biotechnology
• /
• 제16권3호
• /
• pp.469-474
• /
• 2006

Partial nitrification blocking of the oxidation of nitrite ($NO_{2}^{-}$) to nitrate ($NO_{3}^{-}$) has cost-efficient advantages such as lower oxygen and organics demand for nitrification and denitrification, respectively. A nitrifying membrane bioreactor of submerged type was operated for the treatment of synthetic ammonium wastewater with the purpose of nitrite build-up without affecting the efficiency of ammonium oxidation. A high ammonium concentration (1,000 mg/l) was completely converted to nitrate at up to 2 kg $N/m^3$ day under sufficient aeration. The control of pH under sufficient aeration was not a reliable strategy to maintain stable nitrite build-up. When the dissolved oxygen concentration was kept at 0.2-0.4 mg/l by adjusting the aeration rate, about 70% of nitrite content was obtained with ammonium oxidation efficiency higher than 93%. The increase of suction pressure due to membrane fouling was not significant under lowered aerating environment over a 6-month period of operation. The composition of nitrifier community, including relative abundance of nitrite oxidizers in a nitrite-accumulating condition, was quantified by fluorescence in situ hybridization analysis.

• Biomolecules & Therapeutics
• /
• 제24권1호
• /
• pp.33-39
• /
• 2016

Oxidative stress activates several intracellular signaling cascades that may have deleterious effects on neuronal cell survival. Thus, controlling oxidative stress has been suggested as an important strategy for prevention and/or treatment of neurodegenerative diseases. In this study, we found that ginsenoside Rh1 inhibited hydrogen peroxide-induced reactive oxygen species generation and subsequent cell death in rat primary astrocytes. Rh1 increased the expression of phase II antioxidant enzymes, such as heme oxygenase-1 (HO-1), NAD(P)H:quinone oxidoreductase 1, superoxide dismutase-2, and catalase, that are under the control of Nrf2/ARE signaling pathways. Further mechanistic studies showed that Rh1 increased the nuclear translocation and DNA binding of Nrf2 and c-Jun to the antioxidant response element (ARE), and increased the ARE-mediated transcription activities in rat primary astrocytes. Analysis of signaling pathways revealed that MAP kinases are important in HO-1 expression, and act by modulating ARE-mediated transcriptional activity. Therefore, the upregulation of antioxidant enzymes by Rh1 may provide preventive therapeutic potential for various neurodegenerative diseases that are associated with oxidative stress.

• Food Science and Biotechnology
• /
• 제16권4호
• /
• pp.655-658
• /
• 2007

Bacillus polyfermenticus SCD was transformed by the recombinant shuttle vector for Bacillus and Escherichia coli containing 3 antibiotic resistant genes and an ${\alpha}$-amylase gene from Bifidobacterium adolescentis Int57. The ${\alpha}$-amylase gene fused to a secretion sequences was expressed under the control of the promoter of amylase gene from B. subtilis var. natto. The recombinant plasmid was maintained stably in the transformants producing the ${\alpha}$-amylase. The enzyme was secreted to outside of the cell and showed the similar enzyme activity as that of Bacillus subtilis BD170 under the same conditions of pH and growth temperature. Because of the relatively easy transformation and the secretion of the enzyme, the transformants of B. polyfermenticus SCD may give a new strategy in the production of foreign genes.

• 대한전기학회:학술대회논문집
• /
• 대한전기학회 2002년도 학술대회 논문집 전문대학교육위원
• /
• pp.55-59
• /
• 2002

This paper presents a novel three-phase current-fed Pulse Width Modulation converter with switched capacitor type resonant DC link commutation circuit operating PWM pattern strategy under a design consideration of low-pass filter, which can operate on the basis of the principle of zero current soft switching commutation. In the first place, the steady state operating principle of this converter with a new resonant DC link snubber circuit is described in connection with the equivalent operation circuit, together with the practical design procedure of the switched-capacitor type resonant DC link circuit is discussed from a theoretical viewpoint on the basis of a design example for high-power applications. The actively delayed time correction method to compensate distorted currents due to a relatively long resonant commutation time is newly implemented in the open loop control scheme so as to acquire the new optimum PWM pattern. Finally, the experiment of set-up in laboratory system of this converter is concretely demonstrated herein to confirm a zero current soft-switching commutation of this converter. The comparative evaluations between current -fed hard switching PWM and soft-switching PWM converters are carried out from a viewpoint of their PWM converter characteristics.

• 현장농수산연구지
• /
• 제19권1호
• /
• pp.139-151
• /
• 2017

본 연구에서는 기능성 우유의 생산을 위해 높은 항산화 효과를 가지고 심혈관계 질환 환자에게 유익한 CoQ10을 홀스타인 젖소의 우유에 전이시키기 위해 CoQ10 생산성이 높은 R. sphaeroides가 반추위 내에서 생존하며 반추위 환경에 적응하여 CoQ10 생산의 가능성과 반추위 발효에 미치는 영향을 판단하였다. 반추위 환경과 가장 유사한 환경을 만들 수 있는 반추위연속배양장치(RSCC)를 이용하여 CoQ10 생산과 반추 발효를 분석하였다. 전체 15일의 배양기간에서 배양이 안정되는 12일 후부터 5% R. sphaeroides 처리한 군과 R. sphaeroides를 처리하지 않은 대조구로 나누어 실험을 진행한 결과, RSCC system시험에서는 NDF, ADF, pH, NH₃-N그리고 미생물 단백질 합성량은 대조구와 유사한 결과를 보였으나, R. sphaeroides가 성장함으로써 반추위에서 분해되지 않는 반추위 비분해성 단백질인 UDP가 대조구에 유의적으로 높게 나타났다. 특히, CoQ10은 대조구에서는 검출되지 않았으나, 처리구에서는 CoQ10 함량이 336.0 mg/l로 높게 나타났다. 이 연구를 통해서 R. sphaeroides는 반추위 내에서 항산화 물질인 CoQ10을 높은 농도로 생산할 수 있음을 확인하였다. 추후 젖소 착유 실험을 통한 CoQ10 우유 전이 실험을 진행하여 기능성 우유 생산을 위한 첨가제 개발에 활용할 예정이다.

• Journal of Veterinary Science
• /
• 제25권3호
• /
• pp.43.1-43.13
• /
• 2024

Importance: Haemaphysalis longicornis is an obligate blood-sucking ectoparasite that has gained attention due its role of transmitting medically and veterinary significant pathogens and it is the most common tick species in Republic of Korea. The preferred strategy for controlling ticks is a multi-antigenic vaccination. Testing the efficiency of a combination antigen is a promising method for creating a tick vaccine. Objective: The aim of the current research was to analyze the role of subolesin and enolase in feeding and reproduction of H. longicornis by gene silencing. Methods: In this study, we used RNA interference to silence salivary enolase and subolesin in H. longicornis. Unfed female ticks injected with double-stranded RNA targeting subolesin and enolase were attached and fed normally on the rabbit's ear. Real-time polymerase chain reaction was used to confirm the extent of knockdown. Results: Ticks in the subolesin or enolase dsRNA groups showed knockdown rates of 80% and 60% respectively. Ticks in the combination dsRNA (subolesin and enolase) group showed an 80% knockdown. Knockdown of subolesin and enolase resulted in significant depletion in feeding, blood engorgement weight, attachment rate, and egg laying. Silencing of both resulted in a significant (p < 0.05) reduction in tick engorgement, egg laying, egg hatching (15%), and reproduction. Conclusions and Relevance: Our results suggest that subolesin and enolase are an exciting target for future tick control strategies.

• Bulletin of the Korean Chemical Society
• /
• 제26권11호
• /
• pp.1653-1668
• /
• 2005

Assembly of hybrid mesophases through the combination of amphiphilic block copolymers, acting as structuredirecting agents, and silicon sources using low acid catalyst concentration regimes is a versatile strategy to produce large quantities of high-quality ordered large-pore mesoporous silicas in a very reproducible manner. Controlling structural and textural properties is proven to be straightforward at low HCl concentrations with the adjustment of synthesis gel composition and the option of adding co-structure-directing molecules. In this account, we illustrate how various types of large-pore mesoporous silica can easily be prepared in high phase purity with tailored pore dimensions and tailored level of framework interconnectivity. Silica mesophases with two-dimensional hexagonal (p6mm) and three-dimensional cubi (Fm$\overline{3}$m, Im$\overline{3}$m and Ia$\overline{3}$d) symmetries are generated in aqueous solution by employing HCl concentrations in the range of 0.1−0.5 M and polyalkylene oxide-based triblock copolymers such as Pluronic P123 $(EO_{20}-PO_{70}-EO_{20})$ and Pluronic F127 $(EO_{106}-PO_{70}-EO_{106})$. Characterizations by powder X-ray diffraction, nitrogen physisorption, and transmission electron microscopy show that the mesoporous materials all possess high specific surface areas, high pore volumes and readily tunable pore diameters in narrow distribution of sizes ranging from 4 to 12 nm. Furthermore, we discuss our recent advances achieved in order to extend widely the phase domains in which single mesostructures are formed. Emphasis is put on the first synthetic product phase diagrams obtained in $SiO_2$-triblock copolymer-BuOH-$H_2O$ systems, with tuning amounts of butanol and silica source correspondingly. It is expected that the extended phase domains will allow designed synthesis of mesoporous silicas with targeted characteristics, offering vast prospects for future applications.