• Journal of Microbiology and Biotechnology
• /
• v.26 no.5
• /
• pp.846-853
• /
• 2016

We displayed four types of Solanum nigrum metallothionein (SMT) for the first time on the surface of Saccharomyces cerevisiae using an α-agglutinin-based display system. The SMT genes were amplified by RT-PCR. The plasmid pYES2 was used to construct the expression vector. Transformed yeast strains were confirmed by PCR amplification and custom sequencing. Surface-expressed metallothioneins were indirectly indicated by the enhanced cadmium sorption capacity. Flame atomic absorption spectrophotometry was used to examine the concentration of Cd²⁺ in this study. The transformed yeast strains showed much higher resistance ability to Cd²⁺ compared with the control. Strikingly, their Cd²⁺ accumulation was almost twice as much as that of the wild-type yeast cells. Furthermore, surface-engineered yeast strains could effectively adsorb ultra-trace cadmium and accumulate Cd²⁺ under a wide range of pH levels, from 3 to 7, without disturbing the Cu²⁺ and Hg²⁺. Four types of surfaceengineered Saccharomyces cerevisiae strains were constructed and they could be used to purify Cd²⁺-contaminated water and adsorb ultra-trace cadmium effectively. The surface-engineered Saccharomyces cerevisiae strains would be useful tools for the bioremediation and biosorption of environmental cadmium contaminants.

• Journal of Sensor Science and Technology
• /
• v.33 no.3
• /
• pp.139-146
• /
• 2024

Surface-enhanced Raman scattering (SERS) enables the detection of various types of π-conjugated biological and chemical molecules owing to its exceptional sensitivity in obtaining unique spectra, offering nondestructive classification capabilities for target analytes. Herein, we demonstrate an innovative strategy that provides significant machine learning (ML)-enabled predictive SERS platforms through surface-engineered graphene via complementary hybridization with Au nanoparticles (NPs). The hybridized Au NPs/graphene SERS platforms showed exceptional sensitivity (10^-7 M) due to the collaborative strong correlation between the localized electromagnetic effect and the enhanced chemical bonding reactivity. The chemical and physical properties of the demonstrated SERS platform were systematically investigated using microscopy and spectroscopic analysis. Furthermore, an innovative strategy employing ML is proposed to predict various analytes based on a featured Raman spectral database. Using a customized data-preprocessing algorithm, the feature data for ML were extracted from the Raman peak characteristic information, such as intensity, position, and width, from the SERS spectrum data. Additionally, sophisticated evaluations of various types of ML classification models were conducted using k-fold cross-validation (k = 5), showing 99% prediction accuracy.

• Journal of Korean Society on Water Environment
• /
• v.27 no.4
• /
• pp.391-396
• /
• 2011

Recently it revealed that urban runoff was one of the major source that contaminates the river, lake and estuary because it contains toxic compounds such as heavy metals and Poly Aromatic Hydrocarbons (PAHs) as well as suspended solids, organic compounds and nutrients. The engineered polymetric media in this research were developed for reducing pollutants in urban runoff and would be used to be charged in the storm water treatment equipment. The engineered media that were composed of the polypropylene was foamed to have the buoyancy and then shattered by mechanical for the efficient filtration. In this study, Spherical Expanded Polypropylene Media (SEPM), Crushed Polypropylene Media (CPM), Large Crushed Expanded Polypropylene Media (LCEPM), Small Crushed Expanded Polypropylene Media (SCEPM) were made from polypropylene. Surface characteristics of the developed media were determined by scanning electron microscopy analyses. Also, removal efficiencies of SS, $COD_{Cr}$ in the artificial road runoff and the bed headloss by media and particle pollutants captured by media were examined. Results on the surface characteristics of media indicated that SCEPM had the largest specific surface area, $0.80m^2/g$, the lowest specific gravity, 0.091, and the biggest porosity, 0.63, because of crushing the media at the process of manufacturing. And the SCEPM's removal efficiencies of TSS and $COD_{Cr}$ in the artificial road runoff were 92.9% and 83.6%, respectively and the headloss of SCEPM was the least of them.

• Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
• /
• v.22 no.2
• /
• pp.105-115
• /
• 2024

In this study, we employed a small-scale experiment to demonstrate the introduction of a thin copper heat dissipation plate into a bentonite buffer layer of an engineered barrier system. This experiment designed for spent nuclear fuel disposal can effectively reduce the maximum temperature of the bentonite buffer layer, and ultimately, make it possible to reduce the area of the disposal site. For the experiment, a small-scale engineered barrier system with a copper heat dissipation plate was designed and manufactured. the thickness of the cylindrical buffer was about 2 cm, which was about 1/20 of KAERI Repository System (KRS). At a power supply of 250 W, the maximum buffer temperature reduced to a mere 1.8℃ when the thin copper plate was introduced. However, the maximum surface temperature reduced to a remarkable 9.1℃, when a U-collar copper plate was introduced, which had a good contact with the other barrier layers. Consequently, we conclude that the introduction of the thin copper plate into the engineered barrier system for spent nuclear fuel disposal can effectively reduce the maximum buffer temperature in high-level radioactive waste disposal repositories.

• Biomaterials Research
• /
• v.22 no.4
• /
• pp.337-345
• /
• 2018

Background: Human mesenchymal stem cells (hMSCs) are, due to their pluripotency, useful sources of cells for stem cell therapy and tissue regeneration. The phenotypes of hMSCs are strongly influenced by their microenvironment, in particular the extracellular matrix (ECM), the composition and structure of which are important in regulating stem cell fate. In reciprocal manner, the properties of ECM are remodeled by the hMSCs, but the mechanism involved in ECM remodeling by hMSCs under topographical stimulus is unclear. In this study, we therefore examined the effect of nanotopography on the expression of ECM proteins by hMSCs by analyzing the quantity and structure of the ECM on a nanogrooved surface. Methods: To develop the nanoengineered, hMSC-derived ECM, we fabricated the nanogrooves on a coverglass using a UV-curable polyurethane acrylate (PUA). Then, hMSCs were cultivated on the nanogrooves, and the cells at the full confluency were decellularized. To analyze the effect of nanotopography on the hMSCs, the hMSCs were re-seeded on the nanoengineered, hMSC-derived ECM. Results: hMSCs cultured within the nano-engineered hMSC-derived ECM sheet showed a different pattern of expression of ECM proteins from those cultured on ECM-free, nanogrooved surface. Moreover, hMSCs on the nano-engineered ECM sheet had a shorter vinculin length and were less well-aligned than those on the other surface. In addition, the expression pattern of ECM-related genes by hMSCs on the nanoengineered ECM sheet was altered. Interestingly, the expression of genes for osteogenesis-related ECM proteins was downregulated, while that of genes for chondrogenesis-related ECM proteins was upregulated, on the nanoengineered ECM sheet. Conclusions: The nanoengineered ECM influenced the phenotypic features of hMSCs, and that hMSCs can remodel their ECM microenvironment in the presence of a nanostructured ECM to guide differentiation into a specific lineage.

• Tunnel and Underground Space
• /
• v.17 no.6
• /
• pp.464-474
• /
• 2007

An enhancement in the performance and safety of a high-level waste repository requires a validation of its engineered barrier. An engineering-scale test (named "KENTEX") has been conducted to investigate the thermal-hydro-mechanical behaviors in the engineered barrier of the Korean reference disposal system The validation test started on May 31, 2005 and is still under operation. The experimental data obtained allowed a preliminary and qualitative interpretation of the thermal-hydro-mechanical behaviors in the bentonite blocks. The temperature was higher as it became closer to the heater, while it became lower as it was farther away from the heater. The water content had a higher value in the part close to the hydration surface than that in the heater part. The relative humidity data suggested that a hydration of the bentonite blocks might occur by different drying-wetting processes, depending on their position. The total pressure was continuously increased by the evolution of the saturation front in the bentonite blocks and thereby the swelling pressure. Near the heater region, there was also a significant contribution of the thermal expansion of bentonite and the vapor pressure in the pores of the bentonite blocks.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.3
• /
• pp.379-385
• /
• 2014

The purpose of this study was to compare the ability of an engineered Escherichia coli to degrade chlorpyrifos (Cp) using an organophosphorus hydrolase enzyme, encoded in both Flavobacterium sp. ATCC 27551 or Pseudomonas diminuta, by employing the Lpp-OmpA chimera and the N-terminal domain of the ice nucleation protein as anchoring motifs. Tracing of the expression location of the recombinant protein using SDS-PAGE showed the presentation of OPH by both anchors on the outer membrane. This is the first report on the presentation of OPH on the cell surface by Lpp-OmpA under the control of the T7 promoter. The results showed cell growth in the presence of Cp as the sole source of energy, without growth inhibition, and with higher whole-cell activity for both cells harboring plasmids pENVO and pELMO, at approximately 10,342.85 and 10,857.14 U/mg, respectively. Noticeably, the protein displayed by pELMO was lower than the protein displayed by pENVO. It can be concluded that Lpp-OmpA can display less protein, but more functional OPH protein. These results highlight the high potential, of both engineered bacteria, for use in the bioremediation of pesticide-contaminated sources in the environment.

• Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
• /
• v.2 no.1
• /
• pp.23-34
• /
• 2004

To validate the previous conceptual design of cover system, construction of the engineered barrier test facility is completed and the performance tests of the disposal cover system are conducted. The disposal test facility is composed of the multi-purpose working space, the six test cells and the disposal information space for the PR center. The dedicated detection system measures the water content, the temperature, the matric potential of each cover layer and the accumulated water volume of lateral drainage. Short-term experiments on the disposal cover layer using the artificial rainfall system are implemented. The sand drainage layer shows the satisfactory performance as intended in the design stage. The artificial rainfall does not affect the temperature of cover layers. It is investigated that high water infiltration of the artificial rainfall changes the matric potential in each cover layer. This facility is expected to increase the public information about the national radioactive waste disposal program and the effort for the safety of the planned disposal facility.

• Journal of Microbiology and Biotechnology
• /
• v.31 no.9
• /
• pp.1323-1329
• /
• 2021

Micro-scale magnetic beads are widely used for isolation of proteins, DNA, and cells, leading to the development of in vitro diagnostics. Efficient isolation of target biomolecules is one of the keys to developing a simple and rapid point-of-care diagnostic. A zinc finger protein (ZFP) is a double-stranded (ds) DNA-binding domain, providing a useful scaffold for direct reading of the sequence information. Here, we utilized two engineered ZFPs (Stx2-268 and SEB-435) to detect the Shiga toxin (stx2) gene and the staphylococcal enterotoxin B (seb) gene present in foodborne pathogens, Escherichia coli O157 and Staphylococcus aureus, respectively. Engineered ZFPs are immobilized on a paramagnetic bead as a detection platform to efficiently isolate the target dsDNA-ZFP bound complex. The small paramagnetic beads provide a high surface area to volume ratio, allowing more ZFPs to be immobilized on the beads, which leads to increased target DNA detection. The fluorescence signal was measured upon ZFP binding to fluorophore-labeled target dsDNA. In this study, our system provided a detection limit of ≤ 60 fmol and demonstrated high specificity with multiplexing capability, suggesting a potential for development into a simple and reliable diagnostic for detecting multiple pathogens without target amplification.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.10 no.6
• /
• pp.576-581
• /
• 2005

Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. We have engineered the cell surface of the yeast. Saccharomyces cerevisiae, by anchoring active fungal phytase on its cell wall, in order to apply it as a dietary supplement containing bioconversional functions in animal foods and a whole cell bio-catalyst for the treatment of waste. The phytase gene (phyA) of Aspergillus niger with a signal peptide of rice amylase 1A (Ramy1A) was fused with the gene encoding the C-terminal half (320 amino acid residues from the C-terminus) of yeast ${\alpha}-agglutinin$, a protein which is involved in mating and is covalently anchored to the cell wall. The resulting fusion construct was introduced into S. cerevisiae and expressed under the control of the constitutive glyceraldehydes-3-phosphate dehydrogenase (GPD) promoter. Phytase plate assay revealed that the surface-engineered cell exhibited a catalytically active opaque zone which was restricted to the margin of the colony. Additionally, the phytase activity was detected in the cell fraction, but was not detected in the culture medium when it was grown in liquid. These results indicate that the phytase was successfully anchored to the cell surface of yeast and was displayed as its active form. The amount of recombinant phytase on the surface of yeast cells was estimated to be 16,000 molecules per cell.