1.The Effect of Cycolooxygenase (COX-1 and COX-2) on the Bacterial Toxin Formyl Methionyl Leucyl Phenylalanine (fMLP) Induced Contraction of the Rabbit Bladder Muscle.
Suk Young JUNG ; Anthony J BAUER
Korean Journal of Urology 2000;41(2):323-327
No abstract available.
Phenylalanine*
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Urinary Bladder*
2.Approach on the antibacterial activity of some complexes of rare earth elements with L. phenylalanine
Journal of Medical and Pharmaceutical Information 2000;(4):26-28
Antibacterial activity (Staphylococcus aureus and Escherichia coli) of the complexes of La and Eu with L. phenylalanine has proved that they clearly inhibit the growth of Staphylococcus aureus when their concentration ranges from 1.25% to 2.5% and Escherichia coli when their concentrations ranges from 2.5% to 5%. They may have therapeutic value.
Anti-Bacterial Agents
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phenylalanine
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Pharmaceutical Preparations
3.The Study of DNA Mutations of Phenylketonuria in Koreans.
Su Jung YOO ; Yong Hee HONG ; Yong Wha LEE ; Sung Chul JUNG ; Chang Seok KI ; Dong Hwan LEE
Journal of Genetic Medicine 2008;5(1):26-33
PURPOSE: Phenylketonuria(PKU) is an inborn error of metabolism and a genetic disorder resulting from a deficiency of phenylalanine hydroxylase(PAH) and decreased activity of tetrahydrobiopterin(BH4).In this study the correlation between the DNA mutation and clinical manifestations was investigated and PAH DNA mutations were compared bewteen Asian and Caucasian populations. METHODS: DNA was isolated from peripheral leukocytes. The PAH gene was amplified by Polymerase Chain Reaction(PCR) and the sequence was analyzed with Multiplex Ligation-dependent Probe Amplification(MLPA). RESULTS: We characterized the PAH gene of 102 independent Korean patients with PKU. PAH nucleotide sequence analysis revealed 44 different mutations, including 10 novel mutations comprising 9 missense mutations(N207D, K95del, A447P, G344D, P69S, S391I, A202T, G103S, and I306L) and 1 novel splice-site variant mutation(IVS10-3C>G). R243Q was the most prevalent mutation in this study. A259T has not previously been reported in Asian populations, but we found that this mutation had a frequency of 10.1% in our study. Furthermore, the genotypes of BH4 responsive patients were analyzed and were divided into two groups: BH4 medication-only group and BH4 medication with diet therapy group. In the BH4 medication-only group and BH4 medication with diet therapy group, R241C was the most common mutation. CONCLUSION: Novel mutations in the PAH gene of PKU patients are still being discovered. Additional information as to the frequency of mutations in the tetrahydrobiopterine responsive gene is also accumulating. We anticipate that knowledge of these PKU gene mutations will assist the diagnosis, genetic counseling, and therapeutic treatment of PKU patients in future.
Asian Continental Ancestry Group
;
Base Sequence
;
DNA
;
Genetic Counseling
;
Genotype
;
Humans
;
Leukocytes
;
Phenylalanine
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Phenylalanine Hydroxylase
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Phenylketonurias
4.Fungal and Plant Phenylalanine Ammonia-lyase.
Min Woo HYUN ; Yeo Hong YUN ; Jun Young KIM ; Seong Hwan KIM
Mycobiology 2011;39(4):257-265
L-Phenylalanine is one of the essential amino acids that cannot be synthesized in mammals in adequate amounts to meet the requirements for protein synthesis. Fungi and plants are able to synthesize phenylalanine via the shikimic acid pathway. L-Phenylalanine, derived from the shikimic acid pathway, is used directly for protein synthesis in plants or metabolized through the phenylpropanoid pathway. This phenylpropanoid metabolism leads to the biosynthesis of a wide array of phenylpropanoid secondary products. The first step in this metabolic sequence involves the action of phenylalanine ammonia-lyase (PAL). The discovery of PAL enzyme in fungi and the detection of 14CO2 production from 14C-ring-labeled phenylalanine and cinnamic acid demonstrated that certain fungi can degrade phenylalanine by a pathway involving an initial deamination to cinnamic acid, as happens in plants. In this review, we provide background information on PAL and a recent update on the presence of PAL genes in fungi.
Amino Acids, Essential
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Cinnamates
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Deamination
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Fungi
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Mammals
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Phenylalanine
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Phenylalanine Ammonia-Lyase
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Plants
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Resin Cements
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Shikimic Acid
5.Molecular cloning and characterization of three phenylalanine ammonia-lyase genes from Schisandra chinensis.
San-Peng FAN ; Wei CHEN ; Jiang-Chun WEI ; Xiao-Xu GAO ; Yong-Cheng YANG ; An-Hua WANG ; Gao-Sheng HU ; Jing-Ming JIA
Chinese Journal of Natural Medicines (English Ed.) 2022;20(7):527-536
Phenylalanine ammonia-lyase (PAL), which catalyzes the conversion from L-phenylalanine to trans-cinnamic acid, is a well-known key enzyme and a connecting step between primary and secondary metabolisms in the phenylpropanoid biosynthetic pathway of plants and microbes. Schisandra chinensis, a woody vine plant belonging to the family of Magnoliaceae, is a rich source of dibenzocyclooctadiene lignans exhibiting potent activity. However, the functional role of PAL in the biosynthesis of lignan is relatively limited, compared with those in lignin and flavonoids biosynthesis. Therefore, it is essential to clone and characterize the PAL genes from this valuable medicinal plant. In this study, molecular cloning and characterization of three PAL genes (ScPAL1-3) from S. chinensis was carried out. ScPALs were cloned using RACE PCR. The sequence analysis of the three ScPALs was carried out to give basic characteristics followed by docking analysis. In order to determine their catalytic activity, recombinant protein was obtained by heterologous expression in pCold-TF vector in Escherichia coli (BL21-DE3), followed by Ni-affinity purification. The catalytic product of the purified recombinant proteins was verified using RP-HPLC through comparing with standard compounds. The optimal temperature, pH value and effects of different metal ions were determined. Vmax, Kcat and Km values were determined under the optimal conditions. The expression of three ScPALs in different tissues was also determined. Our work provided essential information for the function of ScPALs.
Cloning, Molecular
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Escherichia coli/metabolism*
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Phenylalanine/metabolism*
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Phenylalanine Ammonia-Lyase/chemistry*
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Recombinant Proteins
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Schisandra/genetics*
6.Analysis of mutations in exon 7 of phenylalanine hydroxylase gene among children with phenylketonuria in Ningxia, China.
Xin-Mei MAO ; Jiang HE ; Yuan LIU ; Xiao-Qiang LI ; Wu-Zhong YU ; Zhi-Hui GAO ; Jing CAI
Chinese Journal of Contemporary Pediatrics 2014;16(3):259-262
OBJECTIVETo investigate the type and frequency of mutations in exon 7 of phenylalanine hydroxylase (PAH) gene among children with phenylketonuria (PKU) in Ningxia, China and to provide a basis for the genetic diagnosis and prenatal diagnosis of PKU in this region.
METHODSDirect sequencing of PCR product was performed to analyze the sequences of exon 7 and its flanking introns of 146 PAH alleles in 73 children with typical PKU (39 cases of Hui nationality and 34 cases of Han nationality) in Ningxia.
RESULTSSix mutations were detected, including R243Q (14.4%), R241C (6.8%), IVS7+2T→A (2.7%), L255S (0.7%), G247V (0.7%), and G247R (0.7%). The overall frequency of mutations (missense mutation and splice site mutation) in exon 7 was 26.0% (38/146). The detection rate of R241C mutation was significantly higher in children of Hui nationality than in children of Han nationality(10% vs 3%; P<0.05).
CONCLUSIONSIn Ningxia, R243Q mutation in exon 7 of PAH gene is most common in children with PKU, followed by R241C. The frequency of R241C mutation in exon 7 of PAH gene varies between children with PKU of Hui and Han nationality.
China ; ethnology ; Exons ; Humans ; Mutation ; Phenylalanine Hydroxylase ; genetics ; Phenylketonurias ; genetics
7.Improved screening efficiency for phenylketonuria using a modified bacterial inhibition assay protocol- Autoclaving the bloodspot.
Carrillo Maria Constancia O. ; Tirona Joy ; Capistrano-Estrada Sylvia ; David-Padilla Carmencita
Acta Medica Philippina 2009;43(2):29-31
The Guthrie bacterial inhibition assay (BIA) tests for elevated phenylalanine (PHE) by measuring B. subtilis growth zone density in an agar medium. Dried blood spots with elevated PHE on initial BIA screening undergo repeat BIA testing and thin-layer chromatography (TLC). Specimens with elevated PHE by TLC or BIA on second-tier testing require recall. To streamline PKU screening and reduce the recall rate, we tested a modified BIA protocol incorporating autoclaving of dried blood spots. Autoclaving improves growth zone appearance and has been previously reported to reduce the number of specimen requiring repeat testing. From June to October 2006, dried blood spot samples with initially elevated PHE were autoclaved at 110°C for 5 min, then retested by BIA. Samples with still-elevated PHE were analyzed by TLC. 1078 of 37,268 samples (2.89%) had initially elevated PHE. After autoclaving, 1036 no longer exhibited elevated PHE decreasing to 42 (0.11%) the number requiring TLC. By comparison, the unmodified algorithm resulted in 3.14% of samples received from July - December 2006 requiring both repeat BIA and TLC testing. We have since modified our PKU screening algorithm to require repeat BIA testing from autoclaved samples prior to TLC analysis. This translates to a significant reduction in time and resources for second-tier testing and follow-up, and prevents stress for the parents of a newborn who would have been recalled unnecessarily.
Agar ; Chromatography, Thin Layer ; Phenylalanine ; Mandatory Testing ; Parents ; Algorithms ; Phenylketonurias
8.Gene cloning and enzymatic activity analysis of phenylalanine ammonia-lyase from Sinopodophyllum hexandrum (Royle) Ying.
Di HU ; Xiaowei LUO ; Yuxian WANG ; Ming GONG ; Zhurong ZOU
Chinese Journal of Biotechnology 2023;39(7):2818-2838
Phenylalanine ammonia-lyase (PAL) is the key entry enzyme of plant phenylpropanoid pathway. It plays an important role in the biosynthesis of podophyllotoxin, an anti-tumor lignan that is currently produced from its main natural source Sinopodophyllum hexandrum (Royle) Ying. In this study, we cloned the gene ShPAL encoding phenylalanine ammonia-lyase by RT-PCR from the root of S. hexandrum ecotype inhabited in the Aba' district, Sichuan, based on its public SRA transcriptome data-package. Bioinformatics analyses showed that the ShPAL-encoded protein is composed of 711 amino acids, contains the conserved domains of PAL, and has the signature motif within the active center of aromatic ammonia-lyases. Moreover, ShPAL protein was predicted to have a secondary structure mainly composed of α-helix and random coil, a typical 'seahorse' shape monomer tertiary structure, and a homologous tetramer three-dimensional structure by Swiss-Modelling. The phylogenetic lineage analysis indicated ShPAL was of the highest sequence identity and the shortest evolutionary distance with the PAL of Epimedium sagittatum from the same Berberidaceae family. Subcellular localization experiments showed that ShPAL protein was mainly distributed in the cytoplasm, despite of a minority on the endoplasmic reticulum membrane. Furthermore, ShPAL protein was recombinantly expressed in Escherichia coli and purified by histidine-tag affinity chromatography. Its enzymatic activity was determined up to 20.91 U/mg, with the optimum temperature of 41 ℃ and pH of 9.0. In contrast, the enzyme activity of its F130H mutant decreased by about 23.6%, yet with the same trends of change with temperature and pH, confirming that phenylalanine at this position does affect the substrate specificity of PAL. Both the wild type and the mutant have relatively poor thermostability, but good pH-stability. These results may help to further investigate the regulatory role of PAL in the process of podophyllotoxin biosynthesis and advance the heterologous synthesis of podophyllotoxin to protect the germplasm resource of S. hexandrum. They also demonstrate that ShPAL has a potential application in biochemical industry and biomedicine.
Phenylalanine Ammonia-Lyase/metabolism*
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Podophyllotoxin
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Phylogeny
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Cloning, Molecular
9.Mutations of the phenylalanine hydroxylase (PAH) gene in Filipino patients with phenylketonuria.
Silao Catherine Lynn T. ; Canson Daffodil M. ; Hernandez Karen N. ; Chiong Mary Anne D. ; Capistrano-Estrada Sylvia ; David- Padilla Carmencita
Acta Medica Philippina 2009;43(2):36-39
INTRODUCTION: Phenylketonuria (PKU), an autosomal recessive metabolic disorder caused by phenylalanine hydroxylase (PAH) deficiency, leads to hyperphenylalaninemia and neurological damage if untreated. This is the first study in the Philippines to identify the disease-causing mutations in the PAH gene of clinically diagnosed Filipino PKU patients.
METHODS: The study included four unrelated PKU patients detected by the Philippine Newborn Screening Program from 1996 to 2008. Plasma amino acid analyses for all patients showed increased phenylalanine and low to normal tyrosine levels consistent with the diagnosis of PKU. Mutations in the PAH gene were identified by genomic DNA extraction from dried blood spots of the patients, PAH exon amplification by polymerase chain reaction and subsequent bi-directional DNA sequence analysis.
RESULTS: All patients presented with significantly elevated phenylalanine levels on bacterial inhibition assay and thin layer chromatography. Urinary pterins confirmed the diagnosis of Tetrahydrobiopterin deficiency in two patients while the other 2 patients had the Classical PKU phenotype. Four previously identified mutations in the PAH gene (p.I65T, p.R413P, p.EX6-96A>G, p.R243Q) were identified in those with Classical PKU.
CONCLUSION: The present results confirm the heterogeneity of mutations at the PAH locus in Filipinos. Neonatal screening and the use of molecular diagnosis significantly aid in the medical management and genetic counseling of patients and their families.
Phenylalanine Hydroxylase ; Phenylalanine ; Neonatal Screening ; Genetic Counseling ; Tyrosine ; Pterins ; Chromatography, Thin Layer ; Philippines ; Phenylketonurias ; Exons ; Sequence Analysis, Dna ; Phenotype ; Dna
10.Consensus statement on dietary treatment and nutritional management for phenylalanine hydroxylase deficiency.
Subspecialty Group of Newborn Screening, Society of Birth Defects Prevention and Control, Chinese Preventive Medicine Association ; Subspecialty Group of Clinical Nutrition, the Society of Pediatrics, Chinese Medical Association ; Subspecialty Committee of Clinical Biochemistry and Genetics, the Society of Medical Genetics, Chinese Medical Doctor Association ; Subspecialty Group of Clinical Genetics, The Society of Adolescent Medicine, Chinese Medical Doctor Association
Chinese Journal of Pediatrics 2019;57(6):405-409