• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.4
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• pp.337-343
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• 2006

Trimethylamine dehydrogenase (TMADH, EC 1.5.99.7), an iron-sulfur flavoprotein that catalyzes the oxidative demethylation of trimethylamine to form dimethylamine and formaldehyde, was purified from Methylophaga sp. strain SK1. The active TMADH was purified 12.3-fold through three purification steps. The optimal pH and temperature for enzyme activity was determined to be 8.5 and $55^{\circ}C$, respectively. The $V_{max}\;and\;K_m$ values were 7.9 nmol/min/mg protein and 1.5 mM. A genomic DNA of 2,983 bp from Methylophaga sp. strain SK1 was cloned, and DNA sequencing revealed the open reading frame (ORF) of the gene coding for TMADH. The ORF contained 728 amino acids with extensive identity (82%) to that of Methylophilus methylotrophus $W_3A_1$.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.36-36
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• 2017

Out of twenty common protein amino acids, there are many kinds of non protein amino acids (NPAAs) that exist as secondary metabolites and exert ecological functions in plants. Mimosine (Mim), one of those NPAAs derived from L. leucocephala acts as an iron chelator and reversely block mammalian cell cycle at G1/S phases. Cysteine (Cys) is decisive for protein and glutathione that acts as an indispensable sulfur grantor for methionine and many other sulfur-containing secondary products. Cys biosynthesis includes consecutive two steps using two enzymes-serine acetyl transferase (SAT) and O-acetylserine (thiol)lyase (OASTL) and appeared in plant cytosol, chloroplast, and mitochondria. In the first step, the acetylation of the ${\beta}$-hydroxyl of L-serine by acetyl-CoA in the existence of SAT and finally, OASTL triggers ${\alpha}$, ${\beta}$-elimination of acetate from OAS and bind $H_2S$ to catalyze the synthesis of Cys. Mimosine synthase, one of the isozymes of the OASTLs, is able to synthesize Mim with 3-hydroxy-4-pyridone (3H4P) instead of $H_2S$ for Cys in the last step. Thus, the aim of this study was to clone and characterize the cytosolic (Cy) OASTL gene from L. leucocephala, express the recombinant OASTL in Escherichia coli, purify it, do enzyme kinetic analysis, perform docking dynamics simulation analysis between the receptor and the ligands and compare its performance between Cys and Mim synthesis. Cy-OASTL was obtained through both directional degenerate primers corresponding to conserved amino acid region among plant Cys synthase family and the purified protein was 34.3KDa. After cleaving the GST-tag, Cy-OASTL was observed to form mimosine with 3H4P and OAS. The optimum Cys and Mim reaction pH and temperature were 7.5 and $40^{\circ}C$, and 8.0 and $35^{\circ}C$ respectively. Michaelis constant (Km) values of OAS from Cys were higher than the OAS from Mim. Inter fragment interaction energy (IFIE) of substrate OAS-Cy-OASTL complex model showed that Lys, Thr81, Thr77 and Gln150 demonstrated higher attraction force for Cys but 3H4P-mimosine synthase-OAS intermediate complex showed that Gly230, Tyr227, Ala231, Gly228 and Gly232 might provide higher attraction energy for the Mim. It may be concluded that Cy-OASTL demonstrates a dual role in biosynthesis both Cys and Mim and extending the knowledge on the biochemical regulatory mechanism of mimosine and cysteine.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.134-139
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• 2004

Hybrid cluster protein (HCP) was investigated because of its unique iron-sulfur clusters, which have been found in bacteria and archaea. Here, HCP homologous proteins from the third domain, 'eukarya'(3 amitochondriate protozoans, Giardia lamblia, Entamoeba histolytica, and Trichomonas vaginalis), were identified. All three amitochondriate protozoan HCPs (GlHCP, EhHCP, and TvHCP) belonged to Class I on the basis of two key characters, the cysteine spacing, Cys-(Xaa)₂Cys(Xaa)/sub 7-8/-Cys(Xaa)/sub 5/-Cys, and the absence of N-terminal deletion characteristic to the Class III. In phylogenetic analysis performed with amino acid sequences of 3 eukaryal, 5 bacterial, and 4 archaeal HCPs, the maximum likelihood (ML) tree indicated that TvHCP was clustered with Class I HCPs, whereas the other two HCPs (GlHCP and EhHCP) formed an independent clade with a high bootstrapping value (96%) not belonging to any previously recognized HCP class. In spite of the relatively lower bootstrapping value (61%), the position of the new eukaryal GlHCP-EhHCP clade was close to Class I, including the TvHCP, and Classes II and III were closely related with each other. The finding of eukaryal HCPs would help to understand the evolutionary history of HCP.

• BMB Reports
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• v.41 no.2
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• pp.164-169
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• 2008

The tumor suppressor gene p53 regulates apoptotic cell death and the cell cycle. In this study, we investigated the role of p53 in nitric oxide (NO)-induced apoptosis in vascular smooth muscle cells (VSMCs). We found that the NO donor S-nitroso-N-acetyl-penicillamine (SNAP) increased apoptotic cell death in p53-deficient VSMCs compared with wild-type cells. The heme oxygen-ase (HO) inhibitor tin protoporphyrin IX reduced the resistance of wild-type VSMCs to SNAP-induced cell death. SNAP promoted HO-1 expression in both cell types. HO-2 protein was increased only in wild-type VSMCs following SNAP treatment; however, similar levels of HO-2 mRNA were detected in both cell types. SNAP significantly increased the levels of non-heme-iron and dinitrosyl iron-sulfur clusters in wild-type VSMCs compared with p53-deficient VSMCs. Moreover, pretreatment with FeSO4 and the carbon monoxide donor CORM-2, but not biliverdin, significantly protected p53-deficient cells from SNAP-induced cell death compared with normal cells. These results suggest that wild-type VSMCs are more resistant to NO-mediated apoptosis than p53-deficient VSMCs through p53-dependent up-regulation of HO-2.

• Journal of Microbiology and Biotechnology
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• v.33 no.6
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• pp.707-714
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• 2023

Plant-derived insecticide-neonicotinoid insecticides (NIs) played a crucial role in the development of agriculture and food industry in recent years. Nevertheless, synthesis of these nitrogen-containing heterocyclic compounds with an effective and greener routing remains challenging especially to the notion raise of "green chemistry" and "atom economy". While bio-catalyzed methods mediated by nicotinate dehydrogenase (NDHase) then provide an alternative. The current review mainly focuses on the introduction of sources, components, structure, catalytic mechanism and applications of NDHase. Specifically, NDHase is known as nicotinic acid hydroxylase and the sources principally derived from phylum Proteobacteria. In addition, NDHase requires the participation of the electron respiratory chain system on the cell membrane. And the most important components of the electron respiratory chain are hydrogen carrier, which is mainly composed of iron-sulfur proteins (Fe-S), flavin dehydrogenase (FAD), molybdenum binding protein and cytochromes. Heterologous expression studies were hampered by the plasmid and host with high efficiency and currently only Pseudomonas entomophila L48 as well as Comamonas testosterone was successfully utilized for the expression of NDHase. Furthermore, it is speculated that the conjugate and inductive effects of the substituent group at position 3 of the substrate pyridine ring exerts a critical role in the hydroxylation reactions at position 6 concerning about the substrate molecular recognition mechanism. Finally, applications of NDHase are addressed in terms of pesticide industry and wastewater treatment. On conclusion, this critical review would not only deepen our understanding of the theory about NDHase, but also provides the guideline for future investigation of NDHase.

• Journal of the Korean Dietetic Association
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• v.18 no.2
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• pp.97-114
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• 2012

This study was performed to assess the nutritional status and hair mineral content of children with behavioral problems and compare the values with a gender, age-matched control group. The subjects were recruited from S elementary school children in Seoul, Korea. Students scored higher than 60 points were diagnosed with behavioral problems according to the Korean-Child Behavior Checklist. Nutritional assessment of the two groups (behavior problem group: male n=15, female n=24; $9.6{\pm}1.6$ years, control group: male n=16, female n=18; $9.5{\pm}1.9$ years) was performed using a nutritional survey and by measuring hair mineral contents. In the results, food frequency questionnaire analysis showed that the intakes of anchovy (P<0.05), soybean curb (P<0.01), radish (P<0.05), bean sprouts (P<0.05), spinach (P<0.05), carrot (P<0.05), pumpkin (P<0.05), lettuce (P<0.05), cabbage (P<0.01), apple (P<0.05), and milk (P<0.01) were higher in the control group than the behavior problem group, whereas intakes of ramyeon (P<0.05), cookies (P<0.05), and coke (P<0.01) were higher in the behavior problem group than the control group. Intakes of most nutrients such as plant-derived protein (P<0.05), fiber (P<0.05), plant-derived calcium (P<0.05), phosphorus (P<0.05), plant-derived iron (P<0.05), vitamin B2 (P<0.05), vitamin B6 (P<0.05), vitamin C (P<0.01), vitamin E (P<0.05), and folate (P<0.05) were significantly higher in the control group than the behavior problem group. Hair analysis showed that the levels of arsenic (P<0.05), mercury (P<0.001), uranium (P<0.05), iron (P<0.001), boron (P<0.01), and germanium (P<0.001) were lower, but the levels of phosphate (P<0.05), chromium (P<0.001), sodium (P<0.05), and sulfur (P<0.001) were higher in the behavior problem group than the control group. Conclusively, behavioral problems constitute a complicated condition in which nutritional factors may play major roles. However, it is still under investigation as to whether or not modification of dietary habits or nutritional supplementation can improve children's behavior, since symptoms require a broad understanding of the environmental and genetic interactions.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.65 no.2
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• pp.113-123
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• 2020

Global climatic change and increasing climatic instability threaten crop productivity. Due to climatic change, drought stress is occurring more frequently in crop fields. In this study, we investigated the effect of treatment with hydrogen peroxide (H₂O₂) before leaf development on the growth and yield of sorghum for minimizing the damage of crops to drought. To assess the effect of H₂O₂ on the growth of sorghum plant, 10 mM H₂O₂ was used to treat sorghum leaves at the 3-leaf stage during growth in field conditions. Plant height, stem diameter, leaf length, and leaf width were increased by 7.6%, 9.6%, 8.3% and 11.5%, respectively. SPAD value, chlorophyll fluorescence (Fv/Fm), photosynthetic rate, stomatal conductance, and transpiration rate were increased by 3.0%, 4.9%, 26.0%, 23.4% and 12.7%, respectively. The amount of H₂O₂ in the leaf tissue of sorghum plant treated with 10 mM H₂O₂ was 0.7% of the applied amount after 1 hour. The level increased to approximately 1.0% after 6 hours. The highest antioxidant activity measured by the Oxygen Radical Absorbance Capacity assay was 847.3 µmol·g^-1 at 6 hour after treatment. However, in the well-watered condition, the concentration of H₂O₂ in the plant treated by the foliar application of H₂O₂ was 227.8 µmol·g^-1 higher than that of the untreated control. H₂O₂ treatment improved all the yield components and yield-related factors. Panicle length, plant dry weight, panicle weight, seed weight per plant, seed weight per unit area, and thousand seed weight were increased by 8.8%, 18.0%, 24.4%, 24.7%, 29.9% and 7.1%, respectively. Proteomic analysis showed that H₂O₂ treatment in sorghum increased the tolerance to drought stress and maintained growth and yield by ameliorating oxidative stress.