• Analytical Science and Technology
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• v.9 no.1
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• pp.52-61
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• 1996

Detectable levels of several free D-amino acids were found in some food. This was accomplished by using a column switching method. The determination of total amount of D- and L-amino acids was based on an achiral separation with a $C_{18}$ column. The level of D-amino acids to L-amino acids was determined by the column switching system including the postcolumn reaction detection of the amino acids derivatized with O-phthalaldehyde. The chiral separation of the postcolumn detection system was carried out with chiral crown ether column. This system was applied for the micro-determination of D-amino acids in food such as soy sauce, fermented soy bean and beans. It turned out that the sampling process is critical for the trace analysis of D-amino acids under this achiral / chiral coupled-column system. It was found that commercial soy sauce contained 42ppm, conventional soy sauce 102ppm, fermented soy bean 8.34mg per 1g and bean 2.87mg per 1g sample for phenylalanine. D-phenylalanine was found 0.67% in commercial soy sauce, 0.34% in conventional soy sauce, less than 1.81% in fermented soy bean, and Jess than 2.82% in bean.

• Journal of Microbiology and Biotechnology
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• v.8 no.3
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• pp.237-244
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• 1998

A study was carried out to elucidate the relation between the production of flavonol glycosides and the change of phenylalanine ammonia-lyase activity in cell suspension cultures of Ginkgo biloba by the unassisted and synergistic effects of various factors. The quercetin production showed a mixed-growth-associated pattern in cell suspension cultures. Fluorescent light and UV radiation increased phenylalanine ammonia-lyase (PAL) activity, and resulted in the increase of the production of quercetin and kaempferol ten- and four-fold, respectively, as compared to that obtained in the normal culture condition. The cell growth of Ginkgo biloba was enhanced .at higher temperatures whereas the quercetin production was at its maximum at low temperatures. Moreover, the quercetin production was increased by temperature change during the culture period. In particular, the quercetin production was at the highest level when the culture temperature was elevated from $10^{\circ}C\;to\;30^{\circ}C$. The addition of phenylalanine as a precursor in the culture medium stimulated an 8-fold increase in the production of quercetin; the addition of naringenin caused a l0-fold increase. The quercetin production was also greatly increased by feeding enzyme cofactors such as 2-ketoglutarate and ascorbic acid in the culture medium, but specific PAL activity was not increased except with phenylalanine feeding. The synergistic effect of UV radiation and naringenin feeding was observed, resulting in the increase of flavonol glycoside production at a rate higher than in any other case investigated.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.2
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• pp.246-249
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• 2007

DL-phenylalanine (DLPA) is known as an essential amino acid exhibiting opiate capacities by inhibiting the degradation of enkephalins. From the viewpoint of developing a drink containing DLPA, its solubility and stability in aqueous solutions were investigated at some processing conditions such as temperature and pH. When the solubility was analyzed by transmittance at 600 nm, the solutions containing DLPA of 0.1% to 2% showed transmittance over 98% above $60^{\circ}C$ and over 99% at wide range of pH (3.0, 7.0, and 10.0). The valuable stability was also recognized through HPLC analysis for DLPA content, that is, 61-71% DLPA was still remained even after processing at high temperature and wide pH range, indicating the possibility of de-velopment of drink containing DLPA. Among flavors considered for the improvement of consumers' acceptability on drink, orange and amino acid flavors were superior to others. Microbial growth was not detected during 6-week storage after drink preparation.

• YAKHAK HOEJI
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• v.28 no.4
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• pp.197-206
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• 1984

The derivatives of N-(alkylcarbamoyl) amino acid methyl ester, N-(2-chloroethylcarbamoyl)-glycine methyl ester (7a), -valine methyl ester (8a), -phenylalanine methyl ester (9a), N-(N'-methylcarbamoyl)-glycine methyl ester (7b), -valine methyl ester (8b), and-phenylalanine methyl ester (9b), were prepared by reacting the corresponding free amino acid methyl ester (glycine-, valine-, phenylalanine-methyl ester) with isocyanate (R-N=C=O${\cdot};R=Cl-CH_2-CH_2-or\; CH_3-)$. The prepared N-(alkylcarbamoyl) amino acid methyl esters (7,8,9) were treated with $NaNO_2$/98% HCOOH in order to obtain their nitrosoated products, N-(alkyl-N'-nitrosocarbmoyl)amino acid methyl ester. The compound (7,8,9) gave N-(2-chloroethyl-N'-nitrosocarbamoyl)-valine methyl ester (14a),-phenylalanine methyl ester (15a), N-(N'-alkyl-N'-nitrosocarbamoyl)-glycine methyl ester (13b),-valine methyl ester. (14b), and-phenylalanine methyl ester (15b) respectively under the nitrosoation. On the other hand, N-(2-chloroethylcarbamoyl) glycine methyl ester produced N-(2-chloroethylcarbamoyl)-N-nitrosoglycine methyl ester (13a). The inhibitory activity of the prepared N-(alkylcarbamoyl) amino acid methyl ester (7,8,9) and N-(alkyl-N'-nitrosocarbamoyl) amino acid methyl ester (13,14,15) towards the growth of L1210 murine leukemia cells were examined. Among them the compound (14a) and (15a) exhibit excellent activity having $ED_{50}\; to\;be\;1.5{\mu}g/ml\;and\;3.0{\mu}g/ml respectively.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.16 no.2
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• pp.79-85
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• 2016

Purpose: This study aimed to analyze genetic mutations, clinical manifestations, and treatment of patients with benign HPA in Korea. Methods: This case series study involved ten HPA patients who were referred to our hospital because of high phenylalanine concentration. We investigated their demographic features, clinical manifestations, and mutations of the PAH gene through direct DNA sequencing. Results: Among ten patients with benign HPA, two pairs of patients were related (father-daughter, mother-daughter relationship) cases, and all of them showed no specific clinical manifestations or notable past history. Their plasma phenylalanine levels ranged between 1.2 and 4.2 mg/dL. In the tetrahydrobiopterin (BH4) loading test, all patients were nonresponsive to BH4. In the confirmation test of PAH mutation analysis, we identified eleven different alleles out of twelve. The most common allele was R53H (c.158G> A). In addition, two novel PAH gene mutations, V423A (c.1268T>C) and V51A (c.152T>C), were identified. Although the patients did not receive any pharmacologic treatment or continuous phenylalanine restriction dietary therapy, their neurocognitive development was normal. Moreover, on serial outpatient follow-up tests, all patients maintained phenylalanine levels below 6 mg/dL. Conclusion: This study is the first in Korea to analyze benign HPA patients. All patients with benign HPA could maintain phenylalanine levels below 6 mg/dL with normal neurocognitive development, without continuous therapy. Therefore, performing mutation analysis and distinguishing benign HPA from phenylketonuria (PKU) are important to help improve life quality in patients with benign HPA by avoiding unnecessary lifelong therapy.

• Microbiology and Biotechnology Letters
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• v.20 no.1
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• pp.61-67
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• 1992

The synthesis of N-benzyloxycarbonyl-L-aspartyl-L-phenylalanine methyl ester(ZAPM), a precursor of aspartame, from N-benzyloxycarbonyl-L-aspartic acid(Z-Asp) and L-phenylalanine methyl ester hydrochloride(L-PM-HCl) was investigated in ethylacetate-MES buffer two-phase system using thermolysin. In organic two-phase system, the degree of spontaneous hydrolysis of L-PM. HCl was significantly reduced with increasing the volume ratio of organic to aqueous phase. Stability of thermolysin in organic two-phase system was found to be higher than that in MES buffer solution. More than 90% of initial enzyme activity was maintained after 10 days of incubation in case that the volume of organic phase was equal to that of buffer phase, while the half life of thermolysin was about 2 days in aqueous buffer solution. The results of partitioning of substrates and product in organic two-phase system showed that the difference in partition coefficients between substrates and product was maximum at pH 5.5. The optimal pH for 2-APM synthesis in organic two-phase system was found to be 5.5-5.8, which is consistent with the value expected from the partition experiments. As the concentration of substrates was increased the conversion yield of Z-APM was increased with concomitant reduction of L-PMqHC1 hydrolysis. In case that the concentration of L-PM-HCl and Z-Asp were 160 mM and 80 mM respectively, the conversion yield of Z-APM reached 90% after 28 hrs of reaction. The yield obtained at different volume ratio of organic phase compares well with the predicted equilibrium constant in biphasic system.

• Korean Journal of Environmental Agriculture
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• v.18 no.1
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• pp.11-17
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• 1999

This experiment was conducted to select and utilize the tolerant rice varieties for ozone stress through comparing the physiological responses. In SOD activity, Backunchalbyeo which treated once by 0.15 ppm $0_3$, Ilpumbyeo and Inbujinado treated twice showed significantly different SOD activity compared with $non-O_3$ treatment. No significant difference of SOD activity in three-times $0_3$ treated Sangnambatbyeo and Hangangchalbyeo was found as compared to ($non-O_3$ treated)control, POD activity, when Sangnammbatbyeo, Hangsngchalbyeo, Ilpumbyeo and Inbujinado were sequently treated by third ozone, showed no significant difference between $O_3$ treatment and $non-O_3$. In the comparison of amino acid content in rice leaf blade between three times $O_3$ treatment and $non-0_3$ treatment in 3 varieties, phenylalanine content represent as increased quantities. Especially, in three times $0_3$ treatment of Sangnambatbyeo, phenylalanine content(O.66 mg/g) was significantly increased as compared with $non-O_3$. Visual-growing characters of Sangnembatbyeo were significantly different between $0_3$ treatment and non-treatment. However, grain-filling rate showed no difference. SOD and POD related to resistant mechanism were high activity. In addition, phenylalanine known as precursor which is synthesized resistant materials was increased by $0_3$ treatment. These results suggested that Sangnambatbyeo which increased SOD and POD activity, and phenylalanine content under ozone stress could be selected as a tolerant variety.

• Clinical and Experimental Pediatrics
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• v.51 no.5
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• pp.546-550
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• 2008

In this untreated classic phenylketonuria (PKU) case, mental retardation is severe; however, there have been individuals- like the mother of this case- who have escaped mental retardation and all the other potential sequelae of phenylketonuria, despite having high blood phenylalanine levels, and very poor dietary control. It appears that they have nearly normal brain phenylalanine levels despite high blood phenylalanine (Phe) levels. A number of studies have now demonstrated considerable variability in blood vs. brain phenylalanine levels in phenylketonuria patients. Outcome of phenylketonuria appears to be related to brain phenylalanine levels. We report a case of "undiagnosed" maternal phenylketonuria syndrome. A female infant had low birth weight (2,400 g) with microcephaly. We examined her family and discovered that her mother was an undiagnosed phenylketonuria patient with a borderline intelligence quotient (IQ). The infant's sister, six years old, was diagnosed with phenylketonuria at the age of four years was mentally retarded and had received an operation for cleft lip and palate. the sister had also had a low birth weight (2,300 g). Her sister and mother were compound heterozygotes (mother: R243Q/Y325X; sister: Y325X/P407S). The infant and father were heterozygous carriers (baby: R243Q/ -; father: P407S/ - ).

• Asian-Australasian Journal of Animal Sciences
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• v.9 no.6
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• pp.701-703
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• 1996

The influence of phenylalanine (Phe) in the medium on protein synthesis of chicken embryo fibroblasts (CEF) was examined. CEF was derived from 9-d-old embryos by trypsin-EDTA digestion. To examine the deficiency of Phe in the medium, CEF was cultured in Dulbecco's modified Eagle's medium (DMEM) with or without Phe. CEF was also cultured in Dulbecco's phosphate buffered saline (PBS ($Ca^{2+}$, $Mg^{2+}$)) with or without $400{\mu}m$ Phe in order to examine the effect of Phe supplementation. All media were supplemented with 10% (v/v) fetal calf serum. After incubation for 6, 30 and 54 h, protein synthesis was measured by the incorporation of L-[2, $6-^{3}H$] Phe into CEF for further 18 h. Protein synthesis of CEF cultured in DMEM was higher than that in PBS ($Ca^{2+}$, $Mg^{2+}$). High specific radioactivity of Phe due to the low concentration of Phe in the medium resulted in the apparent increase in protein synthesis of CEF. Protein synthesis cultured in PBS ($Ca^{2+}$, $Mg^{2+}$) with Phe did not increase during 72 h of cell culture.

• Bulletin of the Korean Chemical Society
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• v.23 no.1
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• pp.93-97
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• 2002

Poly(ethylene glycol)-phosphatidylethanolamine conjugate (PEG-PE) has been used in preparing longcirculating liposomes. As a substitute for PEG-PE which can also be used in the long-circulating liposome formualtions, but can be prepared more readily with a lower cost, PEG-Phe-Chol was synthesized from PEG, phenylalanine, and cholesterol. The addition of the PEG derivative to distearoylphosphatidylcholine (DSPC) led to the formation of mixed micelles as well as liposomes when the derivative content was 10 mol% or greater. On the other hand, the addition of just 5 mol% PEG-Phe-Chol to dioleoylphosphatidylethanolamine (DOPE) generated mixed micelles as well as liposomes, but the formation of mixed micelles was completely inhibited by the addition of cholesterol. The leakage of entrapped calcein out of DOPE/cholesterol (7/3) liposomes containing 5 mol% PEG-Phe-Chol was about 45% during the incubation time for 24 h in 50% rabbit plasma, which was similar to that of the same liposomes containing 5 mol% PEG-dipalmitoylphosphatidylethanolamine (DPPE) under the identical conditions.