• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.326-336
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• 2016

Saccharomyces cerevisiae is one of the most employed cell factories for the production of bioproducts. Although monomeric hexose sugars constitute the preferential carbon source, this yeast can grow on a wide variety of nitrogen sources that are catabolized through central nitrogen metabolism (CNM). To evaluate the effects of internal perturbations on nitrogen utilization, we characterized strains deleted or overexpressed in GLT1, encoding for one of the key enzymes of the CNM node, the glutamate synthase. These strains, together with the parental strain as control, have been cultivated in minimal medium formulated with ammonium sulfate, glutamate, or glutamine as nitrogen source. Growth kinetics, together with the determination of protein content, viability, and reactive oxygen species (ROS) accumulation at the single cell level, revealed that GLT1 modulations do not significantly influence the cellular physiology, whereas the nitrogen source does. As important exceptions, GLT1 deletion negatively affected the scavenging activity of glutamate against ROS accumulation, when cells were treated with H₂O₂, whereas Glt1p overproduction led to lower viability in glutamine medium. Overall, this confirms the robustness of the CNM node against internal perturbations, but, at the same time, highlights its plasticity in respect to the environment. Considering that side-stream protein-rich waste materials are emerging as substrates to be used in an integrated biorefinery, these results underline the importance of preliminarily evaluating the best nitrogen source not only for media formulation, but also for the overall economics of the process.

• Pediatric Infection and Vaccine
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• v.29 no.2
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• pp.70-76
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• 2022

Coronavirus disease 2019 (COVID-19) in patients with underlying diseases, is associated with high infection and mortality rates, which may result in acute respiratory distress syndrome and death. Mucopolysaccharidosis (MPS) type II is a progressive metabolic disorder that stems from cellular accumulation of the glycosaminoglycans, heparan, and dermatan sulfate. Upper and lower airway obstruction and restrictive pulmonary diseases are common complaints of patients with MPS, and respiratory infections of bacterial or viral origin could result in fatal outcomes. We report a case of COVID-19 in a 16-year-old adolescent with MPS type II, who had been treated with idursulfase since 5 years of age. Prior to infection, the patient's clinical history included developmental delays, abdominal distension, snoring, and facial dysmorphism. His primary complaints at the time of admission included rhinorrhea, cough, and sputum without fever or increased oxygen demand. His heart rate, respiratory rate, and oxygen saturation were within the normal biological reference intervals, and chest radiography revealed no signs of pneumonia. Consequently, supportive therapy and quarantine were recommended. The patient experienced an uneventful course of COVID-19 despite underlying MPS type II, which may be the result of an unfavorable host cell environment and changes in expression patterns of proteins involved in interactions with viral proteins. Moreover, elevated serum heparan sulfate in patients with MPS may compete with cell surface heparan sulfate, which is essential for successful interaction between the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein and the host cell surface, thereby protecting against intracellular penetration by SARS-CoV-2.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.4
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• pp.206-210
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• 2019

Mucopolysaccharidosis type II (MPS II) is a rare X-linked recessive lysosomal storage disease caused by mutation of the iduronate-2-sulfatase gene. The mutation results in iduronate-2-sulfatase deficiency, which causes the progressive accumulation of heparan sulfate and dermatan sulfate in cellular lysosomes. The phenotype, age of onset, and symptoms of MPS II vary; accordingly, the disease can be classified into either the early-onset type or the late-onset type, depending on the age of onset and the severity of the symptoms. In patients with severe MPS II, symptoms typically first appear between 2 and 5 years of age. Patients with severe MPS II usually die in the second decade of life although some patients with less severe disease have survived into their fifth or sixth decade. Here, we report the establishment of a preimplantation genetic diagnosis (PGD) strategy using multiplex nested polymerase chain reaction, direct sequencing, and linkage analysis. Unaffected embryos were selected via the diagnosis of a single blastomere, and a healthy boy was delivered by a female carrier of MPS II. This is the first successful application of PGD in a patient with MPS II in Korea.

• Korean Journal of Plant Resources
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• v.10 no.3
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• pp.221-226
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• 1997

Improvement of seed protein quality might be an essential issus in soybean and would give more profit directly to both farmers and users. This study was carried out to investigate the effects of reduced-S form(s) on seed storage protein components in soybean during seed filling stages. The reduced-S forms during seed fill were sodium thiosulfate, sodium sulfite, sodium sulfide, thioaceteat, $\beta$-mercaptoethanol, thiourea, thiamine-HCI, L-cysteine, L-cystine, and L-methionine. Seed storage protein concentration did not appear to be affected by any reduced-S forms. However, glycinin and $\beta$-conglycinin concentration seemed to be changed greatly by L-methionine. This resulted in the increase in the 11S/7S ratio(3.58). Among the $\beta$-conglycinin, $\beta$-subunit was not accumulated at all. $\alpha$-subunit concentration appeared to be decreased and $\alpha'$-subunit concentration was not altered in comparison with sulfate control. Also, $\beta$-conglycine concentration, especially $\beta$-subunit concentration, tended to be decreased with L-cystine treatment, resulting in an increase in the 11S/7S ratio(1.83). The glycinin concentration tended to be increased at the expense of the decrease in the $\beta$-conglycinin concentration. Therefore, it is suggested that enhancing soybean protein quality would be achieved by improving metabolic pathways of S assimilation in soybean plants during seed filling period under sulfate-sufficient condition.

• Journal of Applied Biological Chemistry
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• v.42 no.4
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• pp.169-174
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• 1999

Arabidopsis AHL gene contains 4 exons encoding a putative protein highly homologous to the yeast salt-sensitive enzyme HAL2, a 3'(2'),5'-bisphosphate nucleotidase involving in reductive sulfate assimilation. AHL cDNA complemented yeast met22 (hal2) mutant. AHL fusion protein expressed in E. coli exhibited $Mg^{2+}$-dependent, 3'-phosphoadenosine 5'-phosphate (PAP)-specific phosphatase activity. $Li^+,\;Na^+,\;K^+$ and $Ca^{2+}$ ions inhibit the enzyme activity by competing with $Mg^{2+}$ for the active site of the enzyme. The enzyme activity was also sensitive to ${\mu}M$ concentrations of toxic heavy metal ions such as $Cd^{2+},\;Cu^{2+}$ and $Zn^{2+}$, but was not recovered by addition of more $Mg^{2+}$ ions, suggesting that these ions inactivate the enzyme with a mechanism other than competition with $Mg^{2+}$ ions. Inhibition of the AHL enzyme activity may result in accumulation of PAP, which is highly toxic to the cell. Thus, the AHL enzyme could be one of the intial targets of heavy metal toxicity in plants.

• Environmental Engineering Research
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• v.25 no.3
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• pp.274-280
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• 2020

The marine sediment sustains from the anoxic condition due to increased nutrients of external sources. The nutrients are liberated from the sediment, which acts as an internal source. In hypoxic environments, anaerobic respiration results in the formation of several reduced matters, such as N₂ and NH₄⁺, N₂O, Fe²⁺, H₂S, etc. The experimental results have shown that nitrogen and sulfur played an influential, notable role in this biogeochemical cycle with expected chemical reductions and a 'diffusive' release of present nutrient components trapped in pore water inside sediment toward the bulk water. Nitate/ammonium, sulfate/sulfides, and ferrous/ferric irons are found to be the key players in these sediment-waters mutual interactions. Organonitrogen and nitrate in the sediment were likely to be converted to a form of ammonium. Reductive nitrogen is called dissimilatory nitrate reduction to ammonium and denitrification. The steady accumulation in the sediment and surplus increases in the overlying waters of ammonium strongly support this hypothesis as well as a diffusive action of the involved chemical species. Sulfate would serve as an essential electron acceptor so as to form acid volatile sulfides in present of Fe³⁺, which ended up as the Fe²⁺ positively with an aid of the residential microbial community.

• Journal of Microbiology and Biotechnology
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• v.4 no.2
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• pp.126-133
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• 1994

A Pseudomonas putida strain able to accumulate high amount of polyesters of medium-chain-length 3-hydroxyalkanoic acids ($PHA_{MCL)$) was isolated from soil in a landfill site using an enrichment technique. Culture condition of the isolated strain for polyester production in a one-step culture was optimized in a mineral-salts medium against pH and concentrations of ammonium sulfate, carbon source(e.g., octanoate), and phosphate. The optimal values for maximal cell growth and PHA accumulation were: pH; 7$\sim$8, $(NH_4)_2SO_4$; 8 mM, octanoate; 40 mM. The optimum temperature was in the range of $20\sim30^{\circ}C$, which was rather broader than in other bacteria. Cell growth was strongly inhibited by the phosphate limitation to less than 1 mM. An increase of phosphate concentration above 1 mM showed little effect on cell growth and polyester accumulation. When the strain was grown on octanoate under this optimized condition it produced 3.4 g dry biomass per liter and yielded 1.7 g PHA per liter amounting to 53 wt% of dry cells. The monomer units composing the polyester synthesized from octanoate were 3-hydroxyoctanoate (3HO), 3-hydroxycaproate (3HC), and 3-hydroxybutyrate (3HB) (85:13:2, mole ratio). Other low linear $C_3\simC_{10}$ monocarboxylic acids were also tested for polyester production.

• Journal of Microbiology and Biotechnology
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• v.8 no.4
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• pp.301-304
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• 1998

Methylobacterium organophilum can use nitrogen in the form of ammonium ions ($($NH_4$)_2SO_4\;and\;NH_4Cl) and from nonammonium sources such as glycine, alanine, peptone, and yeast extract. When potassium was limited, significantly more PHB was produced when the ammonium ion was the nitrogen source rather than a nonammonium form. With ammonium, the amount of PHB produced was 0.50-0.53 g PHB/l or $52.0~53.2\%$ of the dry cell weight. If nitrogen was from a nonammonium source, the respective values were 0.04~0.06 g PHB/1 or $8.1~11.3\%$ of dry cell weight. When ammonium sulfate was the sole source of nitrogen under potassium-limited conditions, cell growth and PHB accumulation increased as the pH increased from 6.0 to 7.5. Cell growth and PHB amount at pH 7.5 were 2.50 g dry cell weight/1 and 1.40 g PHB/1, respectively.

• Korean Journal of Medicinal Crop Science
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• v.16 no.2
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• pp.124-130
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• 2008

This experiment was conducted to investigate the optimum nitrogen fertilization conditions for the production of high-quality Rhodiola rosea L. in forcing culture. Up until 48 kg N/10 a of both urea and ammonium sulfate, dry matter yield of root of Rhodiola rosea L. tended to increase with increase in application rates, however, it decreased thereafter in higher application rates. In the case of urea, the content of salidroside in the root of the Rhodiola rosea L. appeared to decrease rapidly from the application rates of 64 kg N/10 a and over. Meanwhile, the content of salidroside in the root tended to decrease gradually with the application rates exceeding 64 kg N/10 a of ammonium sulfate. The optimum fertilization rates of urea and ammonium sulfate was 45-8-20-10-10 (N-P-K-Ca-Mg) kg/10 a according to the curvilinear regression equation. However, considering the nitrogen accumulation in soil, nitrogen translocation into the plant, and dry matter yield and content of salidroside in the root of Rhodiola rosea L., the optimum fertilization rates of urea and ammonium sulfate would be 40-8-20-10-10 kg/10 a and 35-8-20-10-10 kg/10 a, respectively.

• Journal of the Korean Society for Precision Engineering
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• v.19 no.4
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• pp.93-100
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• 2002

The current from the feed motor of a machine tool contains substantial information about the machining state. There have been many researches that investigated the current as a measure for the cutting farces. However it has been reported that this indirect measurement of the cutting farces from the feed motor current is only feasible in low frequency. In this research, it was presented that the bandwidth of the current monitoring can be expanded to 130 Hz. And the unusual behavior of the current was examined in this bandwidth. The cross-feed directional cutting force influences the machined surface of the workpiece, which makes it necessary to estimate this force to control the roughness of the machined sulfate. The current exists in the stationary feed motor, and it can give the useful information on the quality of the machined surface. But the unpredictable behavior of the current prevents applying the current to prediction of the cutting state. Empirical approach was conducted to resolve the problem. As a result, the current was shown to be related to the accumulation of the accumulation of the infinitesimal rotation of the motor. rotation of the motor. Subsequently the relationship between the current and the cutting force was identified.