• Mass Spectrometry Letters
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• v.12 no.4
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• pp.163-171
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• 2021

Emiliania huxleyi is a marine phytoplankton that plays a critical role in global carbon and sulfur cycling. The genome of E. huxleyi has been sequenced, and an in-depth proteomic profile of this organism has been reported. This study analyzed the phosphoproteome of E. huxleyi and identified its changes under calcium-limited conditions. A TiO₂ microcolumn was used for phosphopeptide enrichment, followed by liquid chromatography-tandem mass spectrometry analysis. Overall, we identified 7,010 phosphorylated sites on 3,355 phosphopeptides associated with 2,929 phosphoproteins in E. huxleyi. Quantitative analysis revealed changes in the phosphoproteome in E. huxleyi when ambient conditions changed to calcium-limited conditions, notably the phosphorylation of some transporters was altered. This study provides an overview of protein phosphorylation in E. huxleyi and paves the way for further investigations of its biological functions.

• Mass Spectrometry Letters
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• v.13 no.4
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• pp.139-145
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• 2022

Protein glycosylation is a common post-translational modification by non-template-based biosynthesis. In fungal biotechnology, which has great applications in pharmaceuticals and industries, the importance of research on fungal glycoproteins and glycans is accelerating. In particular, the importance of quantitative analysis of fungal glycans is emerging in research on the production of filamentous fungal proteins by genetic modification. Reliable mass spectrometry-based techniques for quantitative glycomics have evolved into chemical, enzymatic, and metabolic stable isotope labeling methods. In this study, we intend to expand quantitative glycomics by metabolic isotope labeling of glycans in Aspergillus niger, a filamentous fungus model, by the MILPIG method. We demonstrate that incubation of filamentous fungi in a culture medium with carbon-13 labeled glucose (1-¹³C₁) efficiently incorporates carbon-13 into N-linked glycans. In addition, for quantitative validation of this method, light and heavy glycans are mixed 1:1 to show the performance of quantitative analysis of various N-linked glycans simultaneously. We have successfully quantified fungal glycans by MILPIG and expect it to be widely applicable to glycan expression levels under various biological conditions in fungi.

• Journal of The Korean Society of Grassland and Forage Science
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• v.36 no.3
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• pp.237-242
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• 2016

Copper (Cu) is a necessary microelement for plants. However, high concentrations of Cu are toxic to plants that change the regulation of several stress-induced proteins. In this study, an annealing control primer (ACP) based approach was used to identify differentially expressed Cu-induced genes in alfalfa leaves. Two-week-old alfalfa plants (Medicago sativa L.) were exposed to Cu for 6 h. Total RNAs were isolated from treated and control leaves followed by ACP-based PCR technique. Using GeneFishing ACPs, we obtained several genes those expression levels were induced by Cu. Finally, we identified several genes including UDP-glucuronic acid decarboxylase, transmembrane protein, small heat shock protein, C-type cytochrome biogenesis protein, mitochondrial 2-oxoglutarate, and trans-2,3-enoyl-CoA reductase in alfalfa leaves. These identified genes have putative functions in cellular processes such as cell wall structural rearrangements, transduction, stress tolerance, heme transport, carbon and nitrogen assimilation, and lipid biosynthesis. Response of Cu-induced genes and their identification in alfalfa would be useful for molecular breeding to improve alfalfa with tolerance to heavy metals.

• Food Science and Biotechnology
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• v.15 no.2
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• pp.238-243
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• 2006

The methanol extract of waxy brown rice fermented with Agrocybe cylindracea was prepared. The extract was then freeze dried and fed to rats at the level of 0.5, 1.0, 2.0 g/kg body weight for 14 days, followed by the treatment with carbon tetrachloride for three consecutive days to induce hepatotoxicity. After sacrificing the rats, the enzyme activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and ${\gamma}$-glutamyl transpeptidase (${\gamma}$-GTP) in serum was determined. Biochemical analysis on serum for albumin, total protein, triglyceride, and total as well as HDL-cholesterol were carried out along with a histopathological study of liver tissues. Based on these data, we suggest that the waxy brown rice cultured with A. cylindracea may exert hepatoprotective activity against hepatotoxicity caused by chemicals such as carbon tetrachloride.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.7
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• pp.1010-1016
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• 2000

Effect on the bio-mass and nutrient yield of Lemna perpusilla Torry was studied at two nutrient loading frequencies. Six polythene sinks ($6{\times}2{\times}0.5m^{3}$ area) randomly allocated to two treatments having three sinks in each treatment. Each sink containing $6{\times}2{\times}0.35m^{3}$ water body, was changed either daily with 12.6 kg anaerobically fermented cattle manure (effluent) or with 76 kg efluent at six-day interval. Media Kjeldhal-N (40 vs. 36 mg/L; p>0.05) and $NH_3$-N (9 vs 7 mg/L; p<0.05) concentration were higher in daily charged sinks than those charged at six-day interval. However, temperature ($28.48{^{\circ}C}\;vs.\;28.60{^{\circ}C}$) and pH (6.82 vs. 6.84) were sinilar in both groups of sinks. Biomass (93 vs. $80g/m^{2}/d$) and crude protein (1.24 vs. $1.11g/m^{2}/d$) yield were non-significantly (p>0.05) higher in the daily charged than 6-d interval charged ponds. However, dry matter (4.52 vs. 4.15%), organic matter (83 vs. 84%), crude protein (31.4 va. 29.6%) and organic carbon (47 vs. 47%) content were similar in both daily and 6-d interval charged ponds. There were no apparent relationships between the crude protein content of Lemna and the media Kjeldhal-N or $NH_3$-N concentration apparently due to saturated level of nutrient (N in particular). It was concluded that with the level of effluent used for two treatments, nutrient-loading frequency had no significant effect on the biomass and protein yield of Lemna. Technique described here can be used for year-round duckweed production from fermented cattle manure for feeding poultry/fish in smallholder production system.

• Proceedings of the Microbiological Society of Korea Conference
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• 2008.05a
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• pp.163-163
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• 2008

Methylophaga aminisulfidivorans $MP^T$, a restricted facultative marine methylotrophic bacterium, was able to utilize methanol as a sole carbon and energy source, and possessed a methanol dehydrogenase (MDH) that is a key enzyme in the process of methanol oxidation. During purification of MDH, three types of MDH (MDH I, II, and III) were obtained in the cell free extracts from $MP^T$ cells grown on methanol. When analyzed by SDS-PAGE and ESI-FT ICR MS, MDH I was confirmed to consist of two subunits and with molecular masses of ~66 and ~10 kDa, respectively, in a form of ${\alpha}_2{\beta}_2$. While MDH II and MDH III contained an additional ~30 kDa protein, designated ${\gamma}$, in a form of ${\alpha}_2{\beta}_2{\gamma}$ and ${\alpha}_2{\beta}_2{\gamma}_2$, respectively. MDH III showed 1.5.2.0 times higher activity than MDH II, while MDH I remained the lowest activity. Based on these observations and experimental data, it seems that the original MDH conformation is ${\alpha}_2{\beta}_2{\gamma}2$ within $MP^T$ growing on methanol, and subunit ${\gamma}$ keeps MDH in an active form, and/or makes MDH easily bind to the substrate, methanol.

• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.489-499
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• 2019

Subunit vaccines are safer and more stable than live vaccines although they have the disadvantage of eliciting poor immune response. To develop a subunit vaccine, an effective delivery system targeting the key elements of the protective immune response is a prerequisite. In this study, oxidized carbon nanospheres (OCNs) were used as a subunit vaccine delivery system and tuberculosis (TB) was chosen as a model disease. TB is among the deadliest infectious diseases worldwide and an effective vaccine is urgently needed. The ability of OCNs to deliver recombinant Mycobacterium tuberculosis (Mtb) proteins, Ag85B and HspX, into bone marrow derived macrophages (BMDMs) and dendritic cells (BMDCs) was investigated. For immunization, OCNs were mixed with the two TB antigens as well as the adjuvant monophosphoryl lipid A (MPL). The protective efficacy was analyzed in vaccinated mice by aerosol Mtb challenge with a virulent strain of Mtb and the bacterial burdens were measured. The results showed that OCNs are highly effective in delivering Mtb proteins into the cytosol of BMDMs and BMDCs. Upon immunization, this vaccine formula induced robust Th1 immune response characterized by cytokine profiles from restimulated splenocytes and specific antibody titer. More importantly, enhanced cytotoxic $CD8^+$ T cell activation was observed. However, it did not reduce the bacteria burden in the lung and spleen from the aerosol Mtb challenge. Taken together, OCNs are highly effective in delivering subunit protein vaccine and induce robust Th1 and $CD8^+$ T cell response. This vaccine delivery system is suitable for application in settings where cell-mediated immune response is needed.

• Mycobiology
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• v.49 no.1
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• pp.86-88
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• 2021

The monokaryotic strain, Schizophyllum commune strain IUM1114-SS01, was generated from a basidiospore of dikaryotic parental strain IUM1114. It even showed the decolorizing activities for several textile dyes much better than its parental strain. Based on the results of a single-molecule real-time sequencing technology, we present the draft genome of S. commune IUM1114-SS01, comprising 41.1 Mb with GC contents of the genome were 57.44%. Among 13,380 protein-coding genes, 534 genes are carbon hydrate-active enzyme coding genes.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.5
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• pp.587-602
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• 2011

Improved efficiency in the use of natural resources, pragmatic production systems and environmental sustainability, justified by the need for improved land use systems and increased productivity, are discussed in the context of Asian integrated systems, diversification, and issues of sustainability. The importance of these are reflected by serious inadequate animal protein production throughout Asia, where available supplies cannot match current and projected human requirements up to 2050. Among the ruminant production systems, integrated tree crops-ruminant production systems are grossly underestimated and merit emphasis and expansion. As an example, integrated oil palm- based system is an important pathway for integration with ruminants (buffaloes, cattle, goats and sheep), and provides the entry point for development. The importance and benefits of integrated systems are discussed, involving animals with annual and perennial tree crops, integration with aquaculture, the significance of crop-animal interactions, stratification of the systems, production options, improved use of forages and legumes, potential for enhanced productivity, implications for improved livelihoods of the rural poor and the stability of farm households. The advances in research and development in South East Asia highlight demonstrable increased productivity from animals and meat offtakes, value addition to the oil palm crop, sustainable development, and distinct economic impacts. The results from 12 out of a total of 24 case studies concerning oil palm over the past three decades showed increased yield of 0.49-3.52 mt of fresh fruit bunches (FFB)/ha/yr; increased income by about 30%; savings in weeding costs by 47- 60% equivalent to 21-62 RM/ha/yr; and an internal rate of return of 19% based on actual field data. The results provide important socio-economic benefits for resource-poor small farmers. Potential increased offtakes and additional income exist with the integration of goats. Additionally, the potential for carbon sequestration with tree crops is an advantage. The reasons for low adoption of the syatems are poor awareness of the potential of integrated systems, resistance by the crop- oriented plantation sector, and inadequate technology application. Promoting wider expansion and adoption of the systems in the future is linked directly with coherent policy, institutional commitment, increased investments, private sector involvement, and a stimulus package of incentives.

• Journal of Life Science
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• v.18 no.1
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• pp.84-90
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• 2008

Bacterial colonies which were able to oxidize the manganese were isolated from six soil samples in Byungchon area. Among them, one bacterial strain was selected for this study based on its high manganese oxidation activity. This selected bacterial strain was identified as Pseudomonas sp. MN5 through physiological-biochemical test and analysis of its 16s rRNA sequence. This selected bacterial strain was able to utilize fructose and maltose, but they doesn't utilizing various carbohydrates as a sole carbon source. Pseudomonas sp. MN5 showed a very sensitive to antibiotics such as kanamycin, chloramphenicol, streptomycin and tetracycline, but a high resistance up to mg/ml unit to heavy metals such as lithium, manganese and barium. Optimal manganese oxidation condition of Pseudomonas sp. MN5 was pH 7.5 and manganese oxidation activity was inhibited by proteinase K and boiling treatment. The manganese oxidizing protein produced by Pseudomonas sp. MN5 was purified by ammonium sulfate precipitation, HiTrap Q FF anion exchange chromatography and G3000sw $_{XL}$ gel filtration chromatography. By sodium dodecyl sulfate polyacrylamide gel electrophoresis, three manganese oxidizing protein with estimated molecular weights of 15 kDa, 46.7 kDa and 63.5 kDa were detected. Also, it was estimated that manganese oxidizing protein produced by Pseudomonas sp. MN5 were a kind of porin proteins through internal sequence and N-terminal sequence analysis.