• Microbiology and Biotechnology Letters
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• v.13 no.2
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• pp.119-127
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• 1985

A series of Escherichia coli mutants were exmined for ability to convert glucose and ammonium salts into phenylalanine. This enabled the biochemical changes having major. effects on phenylaianine yield, and interactions between mutations, to be identified. Changes to the common pathway of aromatic biosynthesis having a major effects include desensitization of the first enzyme (3-deoxy-D-arabinoheptulosonate synthase) to end-product inhibition, and removal of repression of enzyme synthesis. It is suggested that the 3-deoxy-D-arabino-heptulosonate synthase Phe isoenzyme has a more important effect on yield. Similarly, removal of repression and end-product inhibition on the phenylalanine terminal pathway increased yield, and changes to both common and branch pathways were synergistic. Blockage of the typrosine and tryptophan pathways had minor effects on phenylalanine yield, and a mutation affecting aramatic amino acid transport (aroP) decreased yield. With multiple-mutation strains hish specific rates of product formation (ie 0.1-0.17g phenylalanine/g cells/h) were obtained.

• Journal of Nutrition and Health
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• v.4 no.4
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• pp.39-46
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• 1971

Since ${\beta}-aminoethylphosphonic$ acid was discovered in the living organism, the biosynthesis and biological function of aminophosphonic acids have been extensively studied. The purpose of this project consists in the two parts: 1)the preparation of DL-1-amino-2-phenylethylphosphonic acid (Phenylalanine aminophosphonic acid) and DL-1-amino-3-methylbutyl-phosphonic acid (Isoleucine aminophosphonic acid) by the method of Chamber and Isbell. 2) the study of metabolism and biological functions of those synthetic materials by the animal experiment (white rats) The importance of this project proved to be the first experience fed by animals for the elucidation of biochemical and metabolic functions in the animal body. The following organic synthesis of DL-1-amino-3-methylbutylphosphonic acid and DL-1-amino-2-phenylethylphosphonic acid are studied. 1)Synthesis of DL-1-amino-3-methylbutylphosphonic acid a) Synthesis of Iso-butylbromide b) Synthesis of Ethyl iso-butylmalonate c) Synthesis of Iso-caproic acid d) Synthesis of $Ethyl-{\alpha}-bromo$ iso-caproate e) Synthesis of $Triethyl-{\alpha}-phosphono$ iso-caproate f) Synthesis of DL-1-amino-3-methylbutylphosphonic acid 2)Synthesis of DL-1-amino-2-phenylethylphosphonic acid a) Synthesis of Diethyl phosphite b) Synthesis of Ethylchloro acetate c) Synthesis of Triethyl phospho acetate d) Synthesis of Triethyl benzyl phospho acetate e) Synthesis of DL-1-amino-2-phenylethylphosphonic acid The synthetic compounds; DL-1-amino-3-methylbutylphosphonic acid and DL-1-amino-2-phenyl ethylphosphonic acid which are essential amino acid (isoleucine, phenylalanine)analogue are supplemented to the animal diet at the level of 0.2% and 0.4% for isoleucine analogue and 0.35% and 0.7% for phenylalanine analogue. The plain isoleucine and phenylalanine at the same level in the diet are fercilitated as comparable groups in this study. Two sets of experience including 100 male rats were carried out for seven weeks each total 14 weeks. During this period, urine samples, and each big organs were collected for the analysis of total nitrogen, phosphorus, and glycogen contents in the individual samples by Micro Kjeldahl Fisk & Subbarow and Nelson Somogye, method. 1) The result of the project a) The yield of DL-1-amino-3-methylbutylphosphonic acid and DL-1-amino-2-phenylethylphosphonic acid showed low tendency at the level of 12.5% and 20% Melting point of those two compounds were very high and the ${\alpha}-amino$ group in the synthetic compounds showed positive reaction with ninhydrin in the violet color. b) Ail the experimental groups included in this study revealed statistically no significant difference in the organ weight, total body nitrogen retention and urinary phosphorus excretion This means isoleucine aminophosphonic acid and Phenylalanine aminophosphonic acid were utilized in the body as much as the plain amino acids, isoleucine and phenylalanine did. c) The glycogen contents in the liver of the phenylalaine aminophosphonic acid gruop showed higher statistically significant(p<0.05) in the comparision with the group of the Phenylalanine and the Standard-2. It was noteworthy that the higher glycogen content in the liver might indicate the significance in the incorporation of phenylalanine aminophosphonic acid into the intermediate of tricarboxylic acid cycle as activated state.

• Journal of Microbiology and Biotechnology
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• v.23 no.8
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• pp.1055-1059
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• 2013

The ectomycorrhizal fungus Tricholoma matsutake grows symbiotically with Pinus densiflora. Phenylalanine ammonia-lyase (E.C. 4.3.1.24) catalyzes the conversion of L-phenylalanine to trans-cinnamic acid. The role of fungal phenylalanine ammonia-lyase, however, has not been clear until now. In this study, the gene encoding phenylalanine ammonia-lyase (PAL), which was isolated from T. matsutake, was cloned and characterized. The PAL gene (tmpal) consists of 2,160 nucleotides, coding for a polypeptide containing 719 amino acid residues. The deduced amino acid sequence of tmpal from T. matsutake shows high identity (70%) with that from Laccaria bicolor. Comparative analysis of the PAL genes among T. matsutake and other species of the class Agaricomycetes showed that both active sites and binding sites were significantly conserved among these genes. The transcriptional analysis of the PAL gene revealed a differential gene expression pattern depending on the developmental stages (mycelium, primordium, stipe, pileus, and gills) of T. matsutake. These results suggest that the PAL gene in T. matsutake plays an important role in multiple physiological functions.

• Biomolecules & Therapeutics
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• v.17 no.1
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• pp.104-110
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• 2009

Phenylketonuria (PKU) is an inherited metabolic disorder caused by mutations in the phenylalanine catabolic enzyme, phenylalanine hydroxylase (PAH). The use of phenylalanine ammonia-lase (PAL) by oral and parenteral routes as a therapeutic drug for PKU has been severely limited due to inactivation by intestinal proteolysis and immune reactions. PEGylation was applied to PAL to reduce the degrees of antigenicity and proteolytic inactivation. Kinetic experiments with native PAL and pegylated PALs were performed, and pH stability, temperature stability, and protease susceptibility were evaluated. Enzyme linked immunosorbent assay (ELISA) was carried out to measure the immune complex between pegylated PALs and antiserum that had been extracted from a PAL-immunized mouse. Pegylated PAL, especially branched pegylated PAL (10 kDa, 1:32), was more active for phenylalanine and more stable in pancreatic proteases than native PAL. Native PAL was optimal at pH 8.5, corresponding to the average pH range of the small intestine; the same finding was noted for pegylated PALs. All linear and branched pegylated PALs had low reactivity with mouse antiserum, especially the 1:16 formulation with linear 5-kDa PEG and the 1:32 formulation with branched 10-kDa PEG. Therefore, we suggest the 1:32 formulation with branched 10-kDa PEG as the most promising formulation for enzyme replacement therapy.

• Proceedings of the Korean Vacuum Society Conference
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• 2011.08a
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• pp.216-216
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• 2011

We will investigate the bonding configurations of phenylalanine and tyrosine adsorbed on the Ge(100) surface using CLPES and DFT calculations. First, the C 1s, N 1s, and O 1s spectra obtained at 300 K revealed that both the amine and carboxyl groups of phenylalanine and tyrosine concurrently participated in adsorption on the Ge(100) surface without bond breaking using CLPES, depending on the extent of coverage. In the second place, we confirmed that the "O-H dissociated-N dative bonded structure" is the most stable structure implying kinetically favorable structure, and the "O-H dissociation bonded structure" is another stable structure manifesting thermodynamically advantageous structure using DFT calculations. This tendency turns up both phenylalanine and tyrosine, similarly. Furthermore, through the CLPES data and DFT calculation data, we discovered that the "O-H dissociated-N dative bonded structure" and the "O-H dissociation bonded structure" are preferred at 0.30 ML and 0.60 ML, respectively. Moreover, we found that the phenyl ring of phenylalanine is located in axial position to Ge(100) surface, but the phenyl ring of tyrosine is located in parallel to Ge(100) surface using DFT calculations.

• Journal of Nutrition and Health
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• v.14 no.2
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• pp.59-70
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• 1981

The objective of this study is to induce the primary characteristics of phenylketonuria in infant rats during the first 2 weeks after birth. The critical biochemical parameter in the development of phenylketonuria is the elevation of plasma phenylalanine while tyrosine is maintained at a relatively low level. A PKU-like condition was induced in infant rats during the first 2 weeks after birth using a modification of our previously published procedure for the development of a temporary (1 to 3 days) PKU-like condition. Phenylalanine was administered by stomach intubation every 6 hours (starting at 6:00 a.m.) at a dose level of 400mg per kg body weight (after birth-day 2 thru 5) and 500mg per kg body weight (day 6 thru 14). Amethopterin was given at 0.00625 or 0.0125mg per kg body weight (day 3 thru 14) and p-chlorophenylalanine at 50 mg per kg body weight (day 5 thru 14) at 9:00 a.m. and 9:00 p.m. At the times measured (6,10 and 14 days) plasma phenylalanine/tyrosine (P/T) ratios were elevated from a normal value of the or less to values ranging from 6 to 15. During the second week after birth a staggering gait, abnormal stance and decreased social behavior were also observed. None of these PKU-like characteristics were apparent in the three control groups receiving (a) no phenylalanine or inhibitors, (b) phenylalanine alone, or (c) inhibitors alone. The establishment of these primary biochemical characteristics of phenylketonuria by stomach intubation of phenylalanine and a combination of low dose levels of enzyme inhibitors to infant rats provides an experimental system which should he valuable for extensive biochemical, histological and behavioral studies in phenylketonuria.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.10
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• pp.1517-1524
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• 2007

The [$^2H_5$]phenylalanine model was compared with the [1-$^{13}C$]leucine method to determine whole body protein synthesis (WBPS) and degradation (WBPD) in sheep fed at two levels. The animals were fed either 103 (M-diet) or 151 (H-diet) kcal $ME/kg^{0.75}/day$ once daily in a crossover design for 21 days each. The isotope dilutions were simultaneously conducted as a primed-continuous infusion of [$^2H_5$]phenylalanine, [$^2H_2$]tyrosine and [1-$^{13}C$]leucine on each dietary treatment. The WBPS and WBPD calculated from the [$^2H_5$]phenylalanine model were lower (p = 0.009 and p = 0.003, respectively) than those calculated from the [1-$^{13}C$]leucine method. The WBPS tended to be higher (p = 0.08) and WBPD was numerically higher (p = 0.33) for H-diet than M-diet in the [$^2H_5$]phenylalanine model, whereas the WBPS was numerically higher (p = 0.37) for H-diet and WBPS remained similar (p = 0.79) between diets in the [1-$^{13}C$]leucine method. However, the absolute values and the directions of WBPS as well as WBPD from M-diet to H-diet were comparable between the [$^2H_5$]phenylalanine model and [1-$^{13}C$]leucine method. Moreover, the values vary depending on the use of the respective amino acid contents in the carcass protein when calculating WBPS and WBPD. Therefore, it is concluded that the [$^2H_5$]phenylalanine model could be used as an alternative to the [1-$^{13}C$]leucine method for the determination of WBPS and WBPD in sheep.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.306-310
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• 2006

For screening of the compounds exhibiting antimicrobial activities against the D-alanyl-D-alanine of Gram positive bacteria, approximately 2,500 actinomycetes isolated from soil were examined far antimicrobial activity. In consequence, we recently isolated the Streptomyces sp. G91353 strain produced an active compound, A91353, that inhibits the growth of Gram positive bacteria. A91353 was identified as N-acetyl-phenylalanine by various spectroscopic methods. The minimum inhibitory concentration (MIC) values of N-acetyl-phenylalanine on Gram positive bacteria such as Streptococcus pyogenes 308A, Streptococcus pyogenes 77A were determined as $50{\mu}g/ml$, respectively, but did not effect on Gram negative strains. These results indicate that N-acetyl-phenylalanine have an antimicrobial activity, which may be caused by the disturbance of D-alanyl-D-alanine synthesis.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.11
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• pp.1770-1778
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• 2020

Objective: This study was conducted to reveal potential metabolic differences of dairy cows fed corn stover (CS) and rice straw (RS) instead of alfalfa hay (AH) as main forage source. Methods: Thirty multiparous mid-late lactation Holstein dairy cows were selected and randomly assigned to three diets, AH, CS, or RS (n = 10). After 13 weeks of the feeding trial, coccygeal arterial and superficial epigastric venous plasma samples were collected before morning feeding for gas chromatography time-of-flight/mass spectrometry analyses. Results: In the artery, 8 and 13 metabolites were detected as differential metabolites between AH and CS, and between AH and RS, respectively. The relative abundance of phenylpropanoate (log₂fold change [FC]) = 1.30, 1.09), panthenol (log₂FC = 2.36, 2.20), threitol (log₂FC = 1.00, 1.07), and 3,7,12-trihydroxycoprostane (log₂FC = 0.79, 0.78) were greater in both CS and RS than in AH, and tyrosine (log₂FC = -0.32), phenylalanine (log₂FC = -0.30), and pyruvic acid (log₂FC = -0.30) were lower in RS than in AH. In the vein, 1 and 7 metabolites were detected as differential metabolites between AH and CS, and between AH and RS, respectively. By comparing AH and RS, we found that metabolic pathways of phenylalanine, tyrosine, and tryptophan biosynthesis and phenylalanine metabolism were enriched by integrative artery and vein analysis. Furthermore, AH and RS, arterial phenylpropanoate and 4-hydroxyproline were positively, and phenylalanine was negatively correlated with milk urea nitrogen. Finally, in AH and CS, arterial panthenol was negatively correlated with feed efficiency. Conclusion: Arterial metabolic profiles changed more than those in the veins from animals on three forage diets, differing in amino acids. We found that phenylalanine, tyrosine, and tryptophan biosynthesis and phenylalanine metabolism were restricted when cows were fed low-quality cereal straw diets.

• Microbiology and Biotechnology Letters
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• v.19 no.5
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• pp.433-438
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• 1991

The plasmid pSY130-14 for the high production of phenylalanine is a temperaturecontrollable expression vector composed of the $P_R$ and the $P_L$ promoter and a temperature sensitive repressor, $cI_{857}$ of bacteriophage lambda. Strain AT2471 harbouring plasmid pSY13O- 14 is induced the phenylalanine production by shifting up the incubation temperaure to $38.5^{\circ}C$. Plasmid stability of E. coli AT2471 harbouring pSY130-14 was very low, it was about 30% after 48 h cultivation at $38.5^{\circ}C$ without kanamycin. The plasmid disappeared immediately at $40^{\circ}C$ without kanamycin, and at $40^{\circ}C$ adding kanamycin, the plasmid stability decreased at the beginning, but rose with the extension of the culture time. For the improvement of plasmid stability, the plasmid obtaind was designated as pSY15O-1 by changing origin region (ori) pACYC 177 of pSY130-14 for ori pSC101. E. coli AT2471 harbouring pSY150-1 was stable at $38.5^{\circ}C$ without tetracycline, and the plasrnid stability was about 40% after 48 h cultivation at $40^{\circ}C$.