• BMB Reports
• /
• v.37 no.3
• /
• pp.292-297
• /
• 2004

The current model for the mechanism of action of the Bacillus thuringiensis Cry $\delta$-endotoxins involves the penetration of the ${\alpha}4-{\alpha}5$ hairpin into the target midgut epithelial cell membranes, followed by pore formation. In this study, PCR-based mutagenesis was employed to identify a critical residue within the ${\alpha}4-{\alpha}5$ loop of the 130-kDa Cry4A mosquito-larvicidal protein. Alanine-substitutions of two charged (Asp-198 and Asp-200) and four polar (Asn-190, Asn-195, Tyr-201 and Tyr-202) residues in the ${\alpha}4-{\alpha}5$ loop were performed. Like the wild-type, all of the mutant toxins were over-expressed as inclusion bodies in Escherichia coli. When E. coli cells expressing each mutant toxin were bioassayed against Aedes aegypti larvae, larvicidal activity was completely abolished for the substitution of only Tyr-202, while replacements at the other positions still retained a high level of toxicity. Further replacement of Tyr-202 with an aromatic side chain, phenylalanine, did not affect the toxicity. These results revealed a crucial role in toxin activity for the conserved aromatic residue at the 202 position within the ${\alpha}4-{\alpha}5$ loop of the Cry4A toxin.

• Korean Journal of Oriental Medicine
• /
• v.11 no.2
• /
• pp.155-165
• /
• 2005

This study was investigated to determine the quality control of Oriental medicine from stores dealing in Oriental medicine around Seoul and Daegu. We tested total 120 samples that widely used 15 species in herbal medicine (Lycii Fructus, Platycodi Radix, Angelicae Gigantis Radix and 12 others) being collected from Oriental medicine clinic, pharmacy, Oriental pharmacy, Oriental medical hospital and Oriental drug store. We have estimated Oriental medicine by K.P. (Korean Pharmacopoeia), K.H.P(Korean Herbal Pharmacopoeia) and announcement of KFDA. The items of examination were identification, purity, loss on drying, ash, acid insoluble ash, extract content, essential oil content, assay, heavy metal limit, and pesticides residue(BHC, DDT, Aldrin, Endrin, Dieldrin). As a result, 20 samples in total 120 samples were not satisfied with the standard and 7 species in total 15 species were not satisfied with the standard. Identification test, extract content test and pesticides residue(BHC, DDT, Aldrin, Endrin, Dieldrin) were satisfied with the standard. The result will be the basic data for the quality control of Oriental medicine.

• Applied Biological Chemistry
• /
• v.36 no.2
• /
• pp.121-126
• /
• 1993

To extract insoluble proteins of sesame meal residue by using microorganism, the sesame meal residue was treated with crude enzyme solution from Bacillus sp. CW-1121. It was found that the solubility reached to maximum at pH 7.5, $45^{\circ}C$. Under optimum condition, the nitrogen solubility with the enzyme solution from Bacillus sp. CW-1121 reached to 60% in 2 hours. Nitrogen solubility of protein from sesame meal showed minimum value at pH 4.5 and significantly increased above pH 6.0. When the protein from sesame meal extracted with Bacillus sp. CW-1121 was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis, water soluble protein was showed 4 bands and salt soluble protein was showed 2 bands. The amino acid composition of water soluble protein, salt soluble protein and free amino acid indicated relatively high contents of serine (17.24 mg/g), glutamic acid (10.77 mg/g) and glutamic acid (6.55 mg/g). Specially, the contents of essential amino acids were high.

• Journal of Plant Biotechnology
• /
• v.43 no.1
• /
• pp.30-36
• /
• 2016

Brassinosteroids (BRs) are essential plant steroid hormones required for cell elongation, plant growth, development and abiotic and biotic stress tolerance. BRs are recognized by BRI1 receptor kinase that is localized in the plasma membrane, and the BRI1 protein will eventually autophosphorylate in the intracellular domain and transphosphorylate BAK1, which is a co-receptor in Arabidopsis thaliana. However, little is known of the role OsBRI1 receptor kinase plays in Oryza sativa, monocotyledonous plants, compared to that in Arabidopsis thaliana, dicotyledonous plants. As such, we have studied OsBRI1 receptor kinase in vitro and in vivo with recombinant protein and transgenic plants, whose phenotypes were also investigated. A OsBRI1 cytoplasmic domain (CD) recombinant protein was induced in BL21 (DE3) E.coli cells with IPTG, and purified to obtain OsBRI1 recombinant protein. Based on Western blot analysis with phospho-specific pTyr and pThr antibodies, OsBRI1 recombinant protein and OsBRI1-Flag protein were phosphorylated on Threonine residue(s), however, not on Tyrosine residue(s), both in vitro and in vivo. This is particularly intriguing as AtBRI1 protein was phosphorylated on both Ser/Thr and Tyr residues. Also, the OsBRI1 full-length gene was expressed in, and rescued, bri1-5 mutants, such as is seen in normal wild-type plants where AtBRI1-Flag rescues bri1-5 mutant plants. Root growth in seedlings decreased in Ws2, AtBRI1, and 3 independent OsBRI1 transgenic seedlings and had an almost complete lack of response to brassinolide in the bri1-5 mutant. In conclusion, OsBRI1, an orthologous gene of AtBRI1, can mediate normal BR signaling for plant growth and development in Arabidopsis thaliana.

• Bulletin of the Korean Chemical Society
• /
• v.34 no.2
• /
• pp.600-608
• /
• 2013

Amide analogs of tridecapeptide ${\alpha}$-factor (WHWLQLKPGQPMYCONH$_2$) of Saccharomyces cerevisiae, in which Trp at position 1 and 3 were replaced with other residues, were synthesized to ascertain whether cooperative interactions between two Trp residues occurred upon binding with its receptor. Analogs containing Ala or Aib at position 3 of the peptide $[Ala_3]{\alpha}$-factor amide (2) and $[Aib_3]{\alpha}$-factor amide (5) exhibited greater decreases in bioactivity than analogs with same residue at position one $[Ala^1]{\alpha}$-factor amide (1) and $[Aib^1]{\alpha}$-factor amide (4), reflecting that $Trp^3$ may plays more important role than $Trp^1$ for agonist activity. Analogs containing Ala or Aib in both position one and three 3, 6 exhibited complete loss of bioactivity, emphasizing both the essential role and the combined role of two indole rings for triggering cell signaling. In contrast, double substituted analog with D-Trp in both positions 9 exhibited greater activity than single substituted analog with D-Trp 8 or deleted analog 7, reflecting the combined contribution of two tryptophane residues of ${\alpha}$-factor ligand to activation of Ste2p through interaction with residue $Tyr^{266}$ and importance of the proper parallel orientation of two indole rings for efficient triggering of signal G protein coupled activation. Among ten amide analogs, $[Ala^{1,3}]{\alpha}$-factor amide (3), $[Aib^{1,3}]{\alpha}$-factor amide (6), [D-$Trp^3]{\alpha}$-factor amide (8) and [des-$Trp^1,Phe^3]{\alpha}$-factor amide (10) were found to have antagonistic activity. Analogs 3 and 6 showed greater antagonistic activity than analogs 8 and 10.

• Proceedings of the Korean Vacuum Society Conference
• /
• 2013.02a
• /
• pp.642-642
• /
• 2013

Graphene, two dimensional single layer of carbon atoms, has tremendous attention due to its superior property such as high electron mobility, high thermal conductivity and optical transparency. Especially, chemical vapor deposition (CVD) grown graphene has been used as a promising material for high quality and large-scale graphene film. Unfortunately, although CVD-grown graphene has strong advantages, application of the CVD-grown graphene is limited due to ineffective transfer process that delivers the graphene onto a desired substrate by using polymer support layer such as PMMA(polymethyl methacrylate). The transferred CVD-grown graphene has serious drawback due to remaining polymeric residues generated during transfer process, which induces the poor physical and electrical characteristics by a p-doping effect and impurity scattering. To solve such issue incurred during polymer transfer process of CVD-grown graphene, various approaches including thermal annealing, chemical cleaning, mechanical cleaning have been tried but were not successful in getting rid of polymeric residues. On the other hand, lithographical patterning of graphene is an essential step in any form of microelectronic processing and most of conventional lithographic techniques employ photoresist for the definition of graphene patterns on substrates. But, application of photoresist is undesirable because of the presence of residual polymers that contaminate the graphene surface consistent with the effects generated during transfer process. Therefore, in order to fully utilize the excellent properties of CVD-grown graphene, new approach of transfer and patterning techniques which can avoid polymeric residue problem needs to be developed. In this work, we carried out transfer and patterning process simultaneously with no polymeric residue by using a metal etch mask. The patterned thin gold layer was deposited on CVD-grown graphene instead of photoresists in order to make much cleaner and smoother surface and then transferred onto a desired substrate with PMMA, which does not directly contact with graphene surface. We compare the surface properties and patterning morphology of graphene by scanning electron microscopy (SEM), atomic force microscopy(AFM) and Raman spectroscopy. Comparison with the effect of residual polymer and metal on performance of graphene FET will be discussed.

• IMMUNE NETWORK
• /
• v.3 no.2
• /
• pp.126-135
• /
• 2003

Background: One of the important factors in the prognosis of chronic hepatitis B patient is the degree of replication of hepatitis B virus (HBV). It has been known that HBV DNA polymerase plays the essential role in the replication of HBV. HBV DNA polymerase is composed of four domains, TP (Terminal protein), spacer, RT (Reverse transcriptase) and RNaseH. Among these domains, tyrosine, the $65^{th}$ residue of TP is an important residue in protein-priming reaction that initiates reverse transcription. If monoclonal antibody that recognizes around tyrosine residue were selected, it could be applied to further study of HBV replication. Methods: To produce TP-specific scFv (single-chain Fv) by phage display, mice were immunized using synthetic TP-peptide contains $57{\sim}80^{th}$ amino acid residues of TP domain. After isolation of mRNA of heavy-variable region ($V_H$) and light-chain variable region ($V_L$) from the spleen of the immunized mouse, DNA of $V_H$ and $V_L$ were obtained by RT-PCR and joined by a DNA linker encoding peptide (Gly4Ser)3 as a scFv DNA fragments. ScFv DNA fragments were cloned into a phagemid vector. scFv was expressed in E.coli TG1 as a fusion protein with E tag and phage gIII. To select the scFv that has specific affinity to TP-peptide from the phage-antibody library, we used two cycles of panning and colony lift assay. Results: The TP-peptide-specific scFv was isolated by selection process using TP-peptide as an antigen. Selected scFv had 30 kDa of protein size and its nucleotide sequences were analyzed. Indirect- and competitive-ELISA revealed that the selected scFv specifically recognized both TP-peptide and the HBV DNA polymerase. Conclusion: The scFv that recognizes the TP domain of the HBV DNA polymerase was isolated by phage display.

• BMB Reports
• /
• v.33 no.2
• /
• pp.172-178
• /
• 2000

The lipoxygenase was purified 35 fold to homogeneity from the Korean red potato by an ammonium sulfate precipitation and DEAE-cellulose column chromatography. The simple purification method is useful for the preparation of pure lipoxygenase. The molecular weight of the enzyme was estimated to be 38,000 by SDS-polyacrylamide gel electrophoreses and Sepharose 6B column chromatography. The purified enzyme with 2 M $(NH_4)_2SO_4$ in a potassium phosphate buffer, pH 7.0, was very stable for 5 months at $-20^{\circ}C$. Because the purified lipoxygenase is very stable, it could be useful for the screening of a lipoxygenase inhibitor. The optimal pH and temperature for lipoxygenase purified from the red potato were found to be pH 9.0. and $30^{\circ}C$, respectively. The Km and Vmax values for linoleic acid of the lipoxygenase purified from the red potato were $48\;{\mu}M$ and $0.03\;{\mu}M$ per minute per milligram of protein, respectively. The enzyme was insensitive to the metal chelating agents tested (2 mM KCN, 1 and 10mM EDTA, and 1 mM $NaN_3$), but was inhibited by several divalent cations, such as $Cu^{++}$, $Co^{++}$ and $Ni^{++}$. The essential amino acids that were involved in the catalytic mechanism of the 5-lipoxygenase from the Korean red potato were determined by chemical modification studies. The catalytic activity of lipoxygenase from the red potato was seriously reduced after treatment with a diethylpyrocarbonate (DEPC) modifying histidine residue and Woodward's reagent (WRK) modifying aspartic/glutamic acid. The inactivation reaction of DEPC (WRK) processed in the form of pseudo-first-order kinetics. The double-logarithmic plot of the observed pseudo-first-order rate constant against the modifier concentration yielded a reaction order 2, indicating that two histidine residues (carboxylic acids) were essential for the lipoxygenase activity from the red potato. The linoleic acid protected the enzyme against inactivation by DEPC(WRK), revealing that histidine and carboxylic amino acids residues were present at the substrate binding site of the enzyme molecules.

• Journal of Animal Environmental Science
• /
• v.8 no.2
• /
• pp.87-98
• /
• 2002

This study was conducted to determine nutritional values of different sources of food residues(FR) released in autumn and to compare them with nutrient requirements on NRC standard feeding system of swine. Hospital or cafeteria FR contained more cooked rice and side dishes residues and less vegetable residues and fruit peel, resulting in higher energy and lower fiber contents, compared to apartment complex FR, which had opposite patterns to these results. Chemical composition between hospital and cafeteria FR was almost similar. Salt(NaCl) content was more than 9 folds of NRC swine requirement, but much lower than the maximum tolerant level. Essenial and non-essential amino acids profile was similar among FR sources. Hospital or cafeteria FR protein had a similar pepsin digestibility to soybean meal protein. Apartment complex FR protein, however, had a much lower pepsin digestibility. When NRC nutrient requirements are considered, FR in swine diets could satisfy requirements of protein and all the essential amino acids, 75${\sim}$111% of digestible or metabolizable energy, and most of the major and minor minerals. All the FR contained extremely low levels of toxic heavy metals, indicating that they are completely safe from these toxic substances. It was concluded that hospital or cafeteria FR could be a nutritionally excellent and balanced feed source for swine.

• Korean Journal of Environmental Agriculture
• /
• v.33 no.2
• /
• pp.144-147
• /
• 2014

BACKGROUND: It is essential to prioritize the exact and clear understanding of agricultural nonpoint source pollution (NPS) controls. The realistic policies and systems should also be developed based on this understanding. Therefore, this study aimed to present agricultural Best Management Practices (BMPs) applicable for the fields based on the Delphi survey result. METHODS AND RESULTS: This study deduced the evaluation items to assess each BMP for agricultural NPS control and conducted the surveying using the Delphi method based on agricultural BMP experts. In addition, its on-the-spot application were evaluated. Considering its importance, technical, social and economic proprieties showed that political support was ranked first and followed by cost investment, labor investment, reduction effect and resident participation. The survey findings by agricultural BMP experts showed the good performance of on-the-spot application can be achieved from fertilization by soil testing, residue and green manure application and contour plowing which are applicable within a field. Agricultural BMPs, highly applicable for the fields, were the countermeasures that farmers who are the principal bodies of agricultural NPS control could be participated directly. CONCLUSION: The active participation of farmers is essential for effective control of agricultural NPS. It is necessary to establish various incentive systems.