• Molecules and Cells
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• v.41 no.4
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• pp.331-341
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• 2018

L-pipecolic acid is a non-protein amino acid commonly found in plants, animals, and microorganisms. It is a well-known precursor to numerous microbial secondary metabolites and pharmaceuticals, including anticancer agents, immunosuppressants, and several antibiotics. Lysine cyclodeaminase (LCD) catalyzes ${\beta}$-deamination of L-lysine into L-pipecolic acid using ${\beta}$-nicotinamide adenine dinucleotide as a cofactor. Expression of a human homolog of LCD, ${\mu}$-crystallin, is elevated in prostate cancer patients. To understand the structural features and catalytic mechanisms of LCD, we determined the crystal structures of Streptomyces pristinaespiralis LCD (SpLCD) in (i) a binary complex with $NAD^+$, (ii) a ternary complex with $NAD^+$ and L-pipecolic acid, (iii) a ternary complex with $NAD^+$ and L-proline, and (iv) a ternary complex with $NAD^+$ and L-2,4-diamino butyric acid. The overall structure of SpLCD was similar to that of ornithine cyclodeaminase from Pseudomonas putida. In addition, SpLCD recognized L-lysine, L-ornithine, and L-2,4-diamino butyric acid despite differences in the active site, including differences in hydrogen bonding by Asp236, which corresponds with Asp228 from Pseudomonas putida ornithine cyclodeaminase. The substrate binding pocket of SpLCD allowed substrates smaller than lysine to bind, thus enabling binding to ornithine and L-2,4-diamino butyric acid. Our structural and biochemical data facilitate a detailed understanding of substrate and product recognition, thus providing evidence for a reaction mechanism for SpLCD. The proposed mechanism is unusual in that $NAD^+$ is initially converted into NADH and then reverted back into $NAD^+$ at a late stage of the reaction.

• KSBB Journal
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• v.7 no.3
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• pp.209-215
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• 1992

The aims of this sutdy was to investgate the action mechanism of polycation on the $\beta$-glucan synthetase II (GS II) related to cell wall synthesis in suspension cultured carrot cells. In the suspension cultured cells treated with poly-L-Iysine($12{\mu}M$) and poly-L-ornithine ($12{\mu}M$) having ploycationic nature, GS II activity increased about 40% and 50% than that of the control respectively. And similar response was observed when ATP and NaF were treated. On the other hand, ploy-L-lysine and ploy-L-ornithine did nor affect the membrane permeability. Phorbol-12-myrlstate-13-acetate (TPA), activator of protein klnase, increased about 35% and 1-(5-isoquinolinyl sulfonyl)-2-methyl-piperrazine (H-7), inhibitor of protein kinase, decreased about 30% of GSII activity than that of control. These results suggest that polycation plays a role in the cell wall synthesis by increasing GS II activity through phosphorylation.

• Journal of Nutrition and Health
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• v.8 no.2
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• pp.15-18
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• 1975

In man, ornithine carbamyl transferase is localized mainly in the liver and to a lesser extent in the smll intestine. Because of these findings, one might expect damage only to the liver or perhaps the small intestine to cause elevated ornithine carbamyl transferase activity in serum $(_S-OCT)$. Serum ornithine carbamyl transferase activity was determined in thirty normal subjects, and fifteen patients with viral hepatitis and fifteen patients with liver cirrhosis. The correlation of $_S-OCT$ with $_S-GPT$ and $_S-GOT$ was studied. The mean $_S-OCT$ activity in thirty normal Korean subjects was $0.094{\pm}0.043$I.U./L of serum. Elevated $_S-OCT$ activity was observed in patients with liver disease. The correlation coefficient was 0.18 for $_S-OCT$ and $_S-GPT$, ana 0.40 for $_S-OCT$ and $_S-GOT$.

• Applied Biological Chemistry
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• v.38 no.2
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• pp.118-122
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• 1995

Escherichia Coli ornithine transcarbamylase is the enzyme which catalyzes the L-citrulline biosynthesis from L-ornithine and carbamyl phosphate. To facilitate the purification of enzyme which will be used for many biochemical studies such as structure and function relationships and catalytic mechanisms, the cloning and expression of E. coli argI gene for ornithine transcarbamylase was conducted. argI was amplified from genomic DNA of E. coli strain of $DH5{\alpha}$, by polymerization chain reaction (PCR) method. The amplified argI gene was ligated to the prokaryotic expression vector pKK223-3 and used for transformation of E. coli TB2 which was deficient of ornithine transcarbamylase. The over-produced enzyme by the tnansformant was purified by ammonium sulfate fractionation, heat denaturation and affinity chromatography. The result of SDS denaturation gel electrophoresis for the purified enzyme showed a single band of about 38 kDa of ornithine transcarbamylase. Kinetic data for the expressed enzyme gave almost the s?????? values as those of the wild type enzyme. The $k_{cat}$, of the enzyme was $1.0{\times}10^5min^{-1}$, and $K_ms$ for ornithine and carbamyl phosphate were 0.35 mM and 0.06 mM, respectively.

• Journal of Nutrition and Health
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• v.12 no.1
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• pp.31-41
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• 1979

Free amino acid in ethanol extracts and total amino acids in hydrolysates of eleven species of edible mushrooms were analyzed and determinated the contents five kind of new amino acid by means of amino acid autoanalyzer and gas liquid chromatography. The result obtained from this study are as follows. 1) Five kind of new amino acid turned out to be ${\alpha}$-aminobutyric acid, allo-isoleucine, ethanolamine, $\gamma$-aminobutyric acid and ornithine. 2) By means of amino acid autoanalyzer, the monoethanolamine was identified on the chromatogram ahead of alanine, ${\alpha}$-aminobutyric acid between peak of threonine and glycine, allo-isoeleucine between peak of valine and leucine, isoleucine, ${\gamma}$-aminobutyric acid followed by proline between peak of leucine, isoleucine and methionine and ornithine between peak of phenylalanine and tyrosine 3) By means of Gas liquid chromatography, the ${\alpha}$-aminobutyric acid was identified on the chromatogram between peaks of alanine and valine, allo-isoleucine between peaks of methionine and isoleucine, monoethanolamine followed by ${\gamma}$-aminobutyric acid between peaks of phenylalanine and ammonia, ornithine between the peaks of ammonia and lysine. 4) Of five amino acids which were identified, ornithine was the highest of its content in the mushroom extracts, and allo-isoleucine, ethanolamine, and ${\gamma}$-aminobutyric acid came next in decreasing order. 5) Also which were identified, ornithine was the highest of its content in the hydrolysates, and ${\alpha}$-aminobutyric acid, ${\gamma}$-aminobutyric acid, allo-isoleucine came next in decreasing order, ethanol extracts and hydrolysates of Auriculariaauricula-Judae(Fr.) $Qu\acute{e}l$ species didn't contain any of five kind of new amino acid. Ornithine also was the highest in the hydrolysates of ll mushrooms.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.16
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• pp.7031-7038
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• 2015

Background: Arginine may play important roles in tumor progression by providing ornithine for polyamine biosynthesis, required for cell growth. The aim of this work was to determine the expression of arginine metabolic pathway enzymes in head and neck squamous cell carcinoma (HNSCC) in northeast India. Materials and Methods: The expressions of arginase isoforms (ARG1 and ARG2), ornithine aminotransferase (OAT) and ornithine decarboxylase (ODC) were examined in fifty paired HNSCC and adjacent non-tumor tissues by immunohistochemistry. Immunocytochemistry, semiquantitative reverse transcription sq-PCR and quantitative real-time qPCR were used to assess protein and mRNA expressions in peripheral blood of fifty HNSCC patients and hundred controls. Results: ARG1 and ODC protein and mRNA were strongly expressed in peripheral blood from HNSCC patients. No ARG2 expression was observed. In vivo, expression of ARG1, ARG2 and ODC was significantly higher in tumor than in non-tumor tissues. Most tumors expressed low levels of OAT, with no difference in tissues or blood, compared to controls. The absolute extent of maximal ARG1 upregulation with qPCR showed 6.23 fold increase in HNSCC. Conclusions: These findings strongly suggest that in HNSCCs, the ARG1 pathway is stimulated leading to the formation of polyamines as indicated by higher ODC expression, which promote tumor growth.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.339-345
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• 2009

Two lactic acid bacteria (LAB) with high ornithine-producing capacity were isolated from kimchi. Examination of the biochemical features using an API kit indicated that the strains belonged to the members of Weissella genus. They were gram positive, short rod-type bacteria, and able to grow anaerobically with $CO_2$ production. The isolates grew well on MRS broth at $25\sim37^{\circ}C$ and pH of 6.0~7.0. The optimum temperature and pH for growth are $30^{\circ}C$ and pH 6.5. The isolates fermented arabinose, ribose, xylose, glucose but not cellobiose, galactose, raffinose, or trehalsoe. The 16S rDNA sequences of isolates showed 99.6% and 99.7% homology with the Weissella koreensis S5623 16S rDNA (access no. AY035891). They were accordingly identified and named as Weissella koreensis OK1-4 and Weissella koreensis OK1-6, and could produce ornithine from MRS broth supplemented with 1% of arginine at a productivity of 27.01 and 31.41 mg/L/h, respectively. This is the first report on the production of ornithine by the genus Weissella isolated from kimchi.

• Archives of Pharmacal Research
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• v.29 no.3
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• pp.209-212
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• 2006

We investigated the effect of protein extract of Asterina pectinifera on the activity of 4 enzymes that may playa role in adenocarcinoma of the colon: quinone reductase (QR), glutathione Stransferase (GST), ornithine decarboxylase (ODC), and cyclooxygenase (COX)-2. QR and GST activity increased in HT-29 human colon adenocarcinoma cells increased that had been exposed to 4 concentrations of the protein extract (80, 160, 200, and $240{\mu}g/mL$). Additionally, 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ODC activity decreased significantly in cells exposed to the extract in concentrations of $160{\mu}g/mL$ (p<0.05), $200{\mu}g/mL$ (p<0.005), and $240{\mu}g/mL$ (p<0.005). TPA-induced COX-2 activity also decreased in cells exposed to extract concentrations of 10, 20, 40, and $60{\mu}g/mL$. COX-2 expression was also inhibited in cells exposed to this extract. These results suggest that this protein extract of A pectinifera has chemopreventive activity in HT-29 human colon adenocarcinoma cells, and therefore, may have the potential to function as a chemopreventive agent in human colorectal cancer.

• Journal of Mushroom
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• v.15 no.1
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• pp.38-44
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• 2017

Sparassis latifolia (formerly S. crispa) is used in food and nutraceuticals or dietary supplements, as rich in flavor compounds and ${\beta}-glucan$. Some previous studies have reported the effects of mushroom on brain function, including its neuroprotective effect. Thus, for this mushroom to be used as an effective nutraceutical for brain function, it would be desirable for it to contain other compounds such as ${\gamma}-aminobutyric$ acid (GABA) in addition to ${\beta}-glucan$. In this study, the enhancement of growth and GABA production in the mycelium of medicinal and edible mushroom S. latifolia was investigated. Amino acids were added externally as the main source of nutrition, and the effects of amino acids were investigated using liquid medium, specifically amino acid-free potato dextrose broth (PDB). The amino acids added were L-glutamic acid (named PDBG medium) and L-ornithine (named PDBO medium). The growth of mycelia was determined to be $0.9{\pm}0.00g/L$, $2.2{\pm}0.16g/L$, and $1.93{\pm}0.34g/L$ PDBG respectively. The GABA content was $21.3{\pm}0.9mg/100g$ in PDB medium, and it in PDBG 1.4% medium, at $115.4{\pm}30.2mg/100g$. However, the PDBO medium was not effective in increasing the GABA content of mycelia. Amino acids had little effect on the ${\beta}-glucan$ content of mycelia. The ${\beta}-glucan$ content was $39.7{\pm}1.4mg/100mg$, $34.4{\pm}0.2mg/100mg$, and $35.2{\pm}9.2mg/100mg$ in PDB, PDBG 1.8% and PDBO 1.4% media, respectively. Addition of glutamic acid and ornithine positively affected the growth of S. latifolia mycelia, and glutamic acid positively affected GABA production; no degradation of GABA was observed with addition of glutamic acid.

• Applied Biological Chemistry
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• v.47 no.4
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• pp.403-408
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• 2004

We studied active substances like crude cell wall components, crude protein, composing amino acids and free amino acids including orinithine cycle-related amino acids such as asparagine, ornithine and citrullin in fully ripe fruits of Korean native squash, Cucurbita moschata Poir. Crude protein content of 'Jeju 2' was the highest with $2,830\;{\mu}g/g$, while 'Sangju' was the lowest with $1,319\;{\mu}g/g$. Regarding the contents of crude cell wall components, fruit 'Kanghaw' was the highest with 2,961 mg% while 'Namhea' was the lowest with 1,582 mg%. Pectin contents of crude cell wall components were the highest in 'Kanghaw' (2,198 mg%) followed by 'Jeju 2' (2,178 mg%) and 'Jeju l' (1,461 mg%). The main contents of amino acids in squash were glutamic acid, aspartic acid, lysine, leucine and valine, which comprised to be more than 50% of total amino acid contents. Especially, in 'Jeju 2' aspartic acid and threonine were not detected. In fully ripe fruits, a total of 34 kinds of free amino acids were detected including 8 kinds of essential amino acids (histidine, isoleucine, leucine, lysine, phenylalanine, methionine, threonine and valine). More than 50% of the total free amino acids were aspartic acid and asparagine, and also all varieties were detected in ornithine, citrullin, and arginine, which are related to Ornithine cycle. There was a big difference in the contents of arginine in all varieties whereas the contents of ornithine and citrullin were very similar. 'Teaan' 29.34% was 7 times higher than 'Namhea' 4.30% in regards to arginine contents.