• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.40-49
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• 2001

The present work presents a novel approach for the dynamic quantification of respiration rates on a small scale by using lysine-producing Corynebacterium glutamicum ATCC 21253. Cells sampeld from batch cultures at different times were incubated ina 12-ml scale bioreactor equipped with a membrane mass spectrometer. Under dynamic conditions, gas exchange across the gas-liquid phase, specific respiration rates, and RQ values were precisely measured. For this purpose, suitable mass balances were formulated. The transport coefficients for $O_2$ and $CO_2$, crucial for calculating the respiration activity, were determined as $k_La_{O2}=9.18h^{-1}$ and $k_La_{CO2}=5.10h^{-1}$ at 400 rpm. The application of the proposed method to batch cultures of C. glutamicum ATCC 21253 revealed the maximum specific respiration rates of $q_{O2}=8.4\;mmol\;g^{-1}h^{-1}\;and\;q_{CO2}=8.7\;mmol\;g^{-1}h^{-1}$ in the middle of the exponential growth phase after 5 h of cultivation. When the cells changed from growth to lysine production due to the depletion of the essential amino acids theonine, methionine, and leucine, $q_{O2}\;and\;q_{CO2}$ decreased significantly and RQ increased. The respiration data exhibited an excellent agreement with previous cultivations of the strain [13]. This confirms the potential of the developed approach to realistically reflect the metabolic activities of cells at their point of sampling. The short-term influence of added threonine, methionine, and leucine was highest during the shift from growth to lysine production, where $q_{O2}\;and\;q_{CO2}$ increased 50% within one minute after the pulse addition of these compounds. Non-growing, yet lysine-producing cells taken from the end of the batch cultivation revealed no metabolic stimulation with the addition of threonine, methionine, and leucine.

• Journal of Life Science
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• v.10 no.3
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• pp.273-278
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• 2000

The chemical compositions as amino acids, minerals, fatty acids, and total polyphenolic compounds of the seeds of leek (Allium tuberusum) were analyzed. The antioxidative activity of water soluble extract from leek seeds was also tested in DPPH ($\alpha$, $\alpha$ - diphenyl-$\beta$ -picrylhydrazyl) method. The chemical compositions of leek seeds were moisture 4.4%, curde protein 25.7%, crude fat 16.6%, and crude ash 2.9%. Major amino acid compositions were proline 11 g, glutamic acid 4.9 g, arginine 2.1g, aspartic acid 1.6g, leucine 1.3g, valine 1.2 g, and methionine 1.1 g as per 100g. Mineral contents were K 215 ppm, Ca 142 ppm, Fe 124 ppm, and Mg 100 ppm. Major fatty acid compositions were linoleic acid 71.9%, oleic acid 12.7%, palmitic acid 8.6%, and stearic acid 1.4%. The changes of contents in polyphenolic compound from leek seeds caused by heat treatment were also listed in the following order; $20^{\circ}C$(364mg/100g), $40^{\circ}C$(462 mg/100g), and $60^{\circ}C$(551 mg/100g). Antioxidative activity as electron donating ability showed in the following order; 0.05% BHT(butylated hydroxytoluene)(45.6%)>0.05% water-extract(31.3%)>0.1% water extract(30.3%). On the basis of chemical analysis, the leek seedsshowed to have relatively high contents of nutrients as amino acids, minerals, fatty acids.

• Applied Biological Chemistry
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• v.25 no.2
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• pp.75-82
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• 1982

Soybean sprouts treated with growth regulators (IAA and BA) had considerably shorter roots than control. The weight and diameter of treated sprouts were increased by $10{\sim}20%$ and 40%, respectively, compared with control. Crude protein and fat contents of the treated soybean sprouts were higher than those of the control by $5{\sim}10%$ and 10%, respectively. Vitamin C contents of the treated soybean sprout were 38mg%, which corresponds to 70% more vitamin C. Soybean sprouts contained 16 amino acids and the treated sprouts contained more arginine, histidine, lysine and methionine than the control, but significantly less proline. Fatty acid compositions of the treated sprouts and the control were similar, and the ratio unsaturated/saturated fatty acids was about 4.4 in the treated and control sprouts.

• Archives of Pharmacal Research
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• v.20 no.5
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• pp.459-464
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• 1997

The human liver cDNA clone UDPGTh2, encoding a liver UDP-glucuronosyltransferase (UDPGT) was isolated from a .gamma. gt 11 cDNA library by hybridization to mouse transferase cDNA clone, UDPGTm1. UDPGTh2 encoded a 529 amino acid protein with an amino terminus membrane-insertion signal peptide and a carboxyl terminus membrane-spanning region. There were three potential asparagine-linked glycosylation sites at residues 67, 68, and 315. In order to obtain UDPGTh2 protein encoded from cloned human liver UDP-glucuronosyltransferase cDNA, the clone was inserted into the pSVL vector (pUDPGTh2) and expressed in COS 1 cells. The presence of a transferase with Mr-52,000 in transfected cells cultured in the presence of $[^{35}S]$ methionine was shown by immunocomplexed products with goat antimouse transferase IgG and protein A-Sepharose and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The expressed UDPGT was a glycoprotein as indicated by electrophoretic mobility shift in Mr-3,000-4,000 when expressed in the presence of tunicamycin. The extent of glycosylation was difficult to assess, although one could assume that glycosyl structures incorporated at the level of endoplasmic reticulum were always the core oligosaccharides. Thus, it is likely that at least two moieties inserted can account for the shift of Mr-3,000-4,000. This study demonstrates the cDNA and deduced amino acid sequence of human liver UDP-glucuronosyltransferase cDNA, UDPGTh2.

• The Korean Journal of Nuclear Medicine
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• v.38 no.2
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• pp.198-204
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• 2004

Tumor cell proliferation is considered to be a useful prognostic indicator of tumor aggressiveness and tumor response to therapy but in vitro measurement of individual proliferation is complex and tedious work. PET imaging provides a noninvasive approach to measure tumor growth rate in situ. Early approaches have used $^{18}F$-FDG or methionine to monitor proliferation status. These 2 tracers detect changes in glucose and amino acid metabolism, respectively, and therefore provide only an indirect measure of proliferation status. More recent studies have focused on DNA synthesis itself as a marker of cell proliferation. Cell lines and tissues with a high proliferation rate require high rates of DNA synthesis. $[^{11}C]Thymidine$ was the first radiotracer for noninvasive imaging of tumor proliferation. The short half-life of $^{11}C$ and rapid metabolism of $[^{11}C]Thymidine$ in vivo make the radiotracer less suitable for routing use. Halogenated thymidine analogs such as 5-iodo-2-deoxyuridine (IUdR) can be successfully used as cell proliferation markers for in vitro studies because these compounds are rapidly incorporated into newly synthesized DNA. IUdR has been evaluated as a potential in vivo tracer in nuclear medicing but the image qualify and the calculation of proliferation rates are impaired by its rapid in vivo degradation. Hence, the thymidine analog $3'-deoxy-3'-^{18}F-fluorothymidine$ (FLT) was recently introduced as a stable proliferation marker with a suitable nuclide half-life and stable in vivo. $[^{18}F]FLT$ is phosphorylated to 3-fluorothymidine monophosphate by thymidine kinase 1 and reflects thymidine kinase 1 activity in proliferating cell. $[^{18}F]FLT$ PET is feasible in clincal use and well correlates with cellular proliferation. Choline is a precursor for the biosynthesis of phospholipids (in particular, phosphatidylcholine), which is the essential component of all eukaryotic cell membranes and $[^{11}C]choline$, which is a new marker for cellular proliferation.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.12 no.3
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• pp.191-200
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• 1979

For the use of antarctic krill as a fond protein source its compositional characteristics were investigated as the first part of the work includes other subjects such as processing of drill paste, concentrates, and fermented or seasoned product. In general composition of fresh frozen and preboiled frozen krill on board, the contents of crude fat and free amino nitrogen were higher in the former than in the latter which contained a high amount of ash. VBN was rather high as much as 37.6 and $26.4\;mg\%$ in both fresh frozen and preboiled krill. The pH of drill homogenates was 7.1 to 7.2 in both cases. Such a low pH might be attributed to a long term storage and temperature fluctuations during frequent transshipping. The amino acid competition of fresh frozen krill meat showed relatively high amount of glutamic acid, aspartic acid, lysine, proline, and leucine while methionine, histidine, serine, tyrosine, and phenylalanine were lower. Among the essential amino acids lysine and leucine were higher and methionine was lower. In tile composition of free amino acid proline, lysing, arginine, and alanine were higher comparatively to the contents of histidine, aspartic acid, serine, and threonine. It is noteworthy for nutritional qualification that tile essential amino acids particularly as lysine were abundant similarly to that of fishes. Heavy metal contents of krill meat 0.039 to 0.048 ppm as Hg, 0.06 to 0.11 ppm as Pb, less than 0.32 ppm as Zn, 0.008 to 0.012 ppm as Cd, 0.61 to 0.68 ppm as Fe, 0.87 to 1.37 ppm as Cu, and nondetective as Cr. A high Cu content seems to be resulted by tile blood pigment of crustacea. The ratio,1 of edible portion to non-edible portion were 37:63 in fresh frozen and 42:58 in preboiled frozen krill respectively. Release of drip after thawing was more in fresh frozen than in preboiled frozen drill marking $36\%$ and $24\%$ of both respectively.

• Journal of Animal Science and Technology
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• v.45 no.2
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• pp.319-326
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• 2003

Angiotensin-I converting enzyme(ACE)inhibitor was isolated from beef by-products. The beef by- product hydrolysates prepared with various proteases were tested for the inhibitory effects against ACE. The proteases used were proteinase A from bakers yeast, protease type ⅩIII fungal and thermolysin. The maximum inhibitory effect was observed after hydrolysis for 12hrs(beef heart) and 24hrs(beef spleen), respectively. After gel filtration, IC50 value was 0.37mg/ml in beef heart and 1.84mg/ml in beef spleen. After RP-HPLC, the IC50 value of peak 1, peak 2, peak 3 and peak-4 were 0.28mg/ml, 0.26mg/ml, 0.25mg/ml and 0.35mg/ml, respectively. In the results of amino acid composition of peak 1, peak 2, peak 3 and peak 4, it was observed that peak 1 was consisted mainly of glycine and methionine, peak 2 was proline, cystine and methionine, peak 3 was proline and peak 4 was alanine, methionine and leucine. In conclusion, beef heart hydrolysate treated with thermolysin+ proteinase A was shown to have the highest inhibitory effect for 12hrs incubation at 37$^{\circ}C$.

• Korean Journal of Poultry Science
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• v.22 no.3
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• pp.155-160
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• 1995

To evaluate the nutritive values of outdoor mass cultivated Spirulina platensis both chemical analysis and bioassay were carried out using adult cockerels. Blue-green algae, Spirulina platensis contained about 71g /l00g DM of crude protein with balanced amino acid profiles although methionine is liable to he limiting to animals. Compared to fish meal, calcium content and calcium : phosphorus ratio of the Spirulina were not suitable in terms of animal requirements. Reasonable amount of y-linolenic acid(C18: 3 $\omega$6) in Spirulina platensis draws a clinical attention due to its historically recognized pharmacotheraputic functions. Metabolizable energy contents of Spirulina were 3.67 and 3.11 mcal /kg DM for TMEn and AMFn, respectively, which therefore, can he a reliable energy source for poultry. True amino acid availabilities of essential amino acids of Spirulina platensis were higher than 90% for poultry, which is better than comparative ingredient like fish meal. Overall data from both chemical analysis and bioassay demonstrated that the Spirulina platensis could he a favorable protein feedstuffs for poultry.

• Journal of Microbiology and Biotechnology
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• v.11 no.2
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• pp.281-286
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• 2001

Since the number of amino groups which are exposed by deacetylation of acetyl-D-glucosamine influences antimicrobial activity, a chitosan oligosaccharide (COS) derivative by N-conjugation of COS with asparagine, an amino acid with two amino groups, was synthesized and the antimicrobial effect on E. coli growth was compared with other COS derivatives which were N-conjugated with glycine, alanine, aspartic acid, cysteins, an methionine, and unmodified COS. The structure of asparagine N-conjugated COS (Asn-COS) derivative was identified by using a FT-IR, $^{13}C\;FT-NMR$, and an elemental analyzer. The antimicrobial activity of Asn-COS against E. coli growth was significantly improved as compared to the other COS derivatives as well as COS itself. This means that Asn-COS with two positive charges strongly interacts with the carboxyl negative charges on the bacteria cell wall. The results for Asn-COS were as follows: 100% bactericidal activity, 0.002% MIC, and no growth of E. coli during 3 days of culture time, suggesting that Asn-COS may be useful as a new antibiotic agent.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.208-213
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• 2000

Penicillium fellutanum produces a phosphorylated, choline-containing extracellular peptido-polysaccharide, peptidophosphogalactomannan (pPxGM) (8). The $\^$13/C-methyl labeled pPxGM ([methyl-$\^$13/C]pPxGM) was prepared from the cultures supplemented with L-[methyl-$\^$13/C]methionine or [2-$\^$13/C]glycine and was used as a probe to monitor the fate of phosphocholine in this polymer. Addition of purified [methyl-$\^$l3/C]pPxGM to growing cultures in low phosphate medium resulted in the disappearance of [methyl-$\^$13/C]phosphocholine and -N,N'-dimethyl-phosphoethanolamine from the added [methyl-$\^$13/C]pPxGM. Two $\^$l3/C-methyl-enriched cytoplasmic solutes, choline-O-sulfate and glycine betaine, were found in mycelial extracts, suggesting that phosphocholine-containing extracellular pPxGM of P.fellutanum is a precursor of intracellular choline-O-sulfate and glycine betaine and thus of phosphatydilcholine (l0). $\^$13/C-Methyl-labeled cells grown in 3 M NaCl-containing medium showed 2.6- and 22-fold more accumulation of $\^$13/C-methyl labeled choline-O-sulfate and glycine betaine, respectively, originated from the extracellular [$\^$13/C-methyl]pPxGM than those grown without added NaCl. The results suggest that, in addition to glycerol and erythritol, glycine betaine and choline-O-sulfate and thus choline are also osmoprotectants and hence that pPxGM is involved in osmotolerance of this fungus (11). Taken collectively, the $\^$l3/C- and $\^$31/P-NMR analyses of cytosolic solute pools and structural modulation of extracellular pPxGM corresponding to environmental stimuli in P. fellutanum, provided evidence that pPxGM is involved in cellular choline metabolism, osmotolerance, and recycling of metabolites.