• 미생물학회지
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• 제26권3호
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• pp.270-277
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• 1988

Microorganisms isolated from soil (150 strains), fungi (39 strains), yeasts (9 strains) and Streptomyces species (39 strains) were assayed for L-phenylalanine ammonia-lyase(PAL) activity. 17 strains of fungi and 46 strains of soil isolates were proved to produce PAL, Aspergillus panamensis, Penicillium varioti and 11 soil isolates showed comparatively large PAL activity. When PAL activity was assayed with cell-free extracts of these 13 strains and 7 strains of Rhodotorula and Rhodosporidium geni, Rhodosporidium toruloides (IFO 0559) showed the highest PAL activity with 0.333 units per g of the wet cell weight.

• 한국균학회지
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• 제39권3호
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• pp.205-212
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• 2011

진균의 phenylalanine ammonia-lyase(PAL)와 식물의 PAL간 그리고 PAL과 histidine ammonia-lyase (HAL)간의 구조적 특성 관계에 대한 이해를 얻기 위해 옥수수병원균인 깜부기균(Ustilago maydis)의 PAL에 대한 면역학적 분석을 수행하였다. 진균의 PAL 항체와 식물의 PAL 항체를 이용하여 분석하였을 때 실험한 모든 종의 PAL 간에 교차반응이 나타났다. 알팔파 PAL 항체와 포플라 PAL 항체 모두 식물 PAL은 강하게 인식하였으나 진균의 PAL은 약하게 인식하였다. U. maydis PAL은 Rhodotorula glutinis 효모의 PAL만 약하게 인식하였다. U. maydis PAL은 식물의 PAL에 대해서 낮은 친화성을 보였으나 Pseudomonas 세균의 HAL에 대해서는 강한친화성을 보였다. 2종의 식물 PAL 항체들 또한 Pseudomonas 세균의 HAL에 대해 강한 친화성을 보였다. 진균과 식물의 PAL 항체는 PAL 효소의 활성 저해를 나타냈으며 세균의 HAL 효소의 활성에 대해서도 중도적 저해를 나타냈다. 세균의 HAL 항체는 식물과 효모와 Ustilago PAL 중에 Ustilago PAL 활성만 저해하였다. 식물의 PAL과 진균의 PAL 모두 효모의 PAL 활성은 저해하지 못했다. 본 연구는 PAL과 HAL간에 면역학적 관계가 있음을 처음으로 보고한다.

• Journal of Microbiology and Biotechnology
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• 제23권8호
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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.

• 한국식품영양학회지
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• 제9권4호
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• pp.430-433
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• 1996

딸기의 2차대사에 관여하는 phenylalanine ammonia-lyase는 풍미나 색소, 향기 등에 큰 영향을 미치며 숙도 측정지표로 여겨진다. phenylalanine ammonia-lyase는 숙성 전단계에 걸쳐 활성을 나타내기는 하나 완숙기와 과숙기에 그 활성이 뚜렷이 증가하였다. 이는 딸기의 숙성과 더불어 de novo 합성이 되기 때문으로 보인다. 분자량은 겔 크로마토그래피로 260,000으로 나타났다.

• Preventive Nutrition and Food Science
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• 제2권4호
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• pp.354-359
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• 1997

Optimal cultivation conditions for the production of phenylalanine ammonia-lyase(PAL) from Rhodotorula aurantiaca K-505 were selected, and the kinetic parameters of the produced PAL were determined. The most suitable carbon and nitrogen sources were glucose and tryptone, respectively. The strain expressed PAL constituttively when using the optimized semi-complex media. High cell density culture could be critical for maximal production of PAl since the PAL ynthesis was growth associated. maximum PAL activity was observed at initial pH 6.0. although the ll growth was not markedly affected by temperature between 22 and 28$^{\circ}C$, the cells yielded the maximum PAL activity when cultivated at 22$^{\circ}C$. The maximum activity for deamination of L-phenylalnine to trans-cinnamic acid was observed around pH 8.8. The PAL activity gave the maximum at 45$^{\circ}C$, and greatly decreased at higher than 5$0^{\circ}C$. Activation energy({TEX}$E_{a}${/TEX}) calculated from Arrhenius equation was 6.28 kcal/mol in the range of 22$^{\circ}C$ to 4$0^{\circ}C$. A oolf plot showed that the enzyme reaction follows Michaelis-Menten equation, whose {TEX}$K_{M}${/TEX} and {TEX}$V_{max}${/TEX} values were 4.65$\times${TEX}$10^{-3}${/TEX} M and 0.89$\mu$ mol/mg-min respectively.

• Food Science and Biotechnology
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• 제18권4호
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• pp.954-958
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• 2009

Effects of controlled- and uncontrolled-pH operations on phenylalanine ammonia lyase (PAL) production by a recombinant Escherichia coli strain were investigated at uncontrolled-pH ($pH_{UC}$) and controlled-pH ($pH_C$) of 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, and 8.5 in bioreactor systems. The results showed that the recombinant PAL activity was improved significantly by controlled pH strategy. Among the $pH_C$ operations, the highest PAL activities were obtained under $pH_C$ 7.5 strategy where cell mass ($OD_{600\;nm}$) and PAL activity was 1.3 and 1.8 fold higher than those of $pH_{UC}$, respectively. The maximum PAL activity reached 123 U/g. The $pH_C$ 7.5 strategy made recombinant plasmid more stable and therefore allowed easier expression of PAL recombinant plasmid, which increased PAL production. It was indicated that the new approach (controlled-pH strategy) obtained in this work possessed a high potential for the industrial production of PAL, especially in the biosynthesis of L-phenylalanine.

• Biomolecules & Therapeutics
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• 제17권1호
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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.

• 한국자원식물학회지
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• 제23권6호
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• pp.535-540
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• 2010

Phenylalanine ammonia-lyase (PAL), a key enzyme of the phenylpropanoid biosynthesis pathway, is activated by a number of developmental and environmental cues. The coding region of the NtPAL4 gene was 2,154 bp in length, and its deduced protein was composed of 717 amino acids. Sequence analysis of NtPAL4 cDNA from tobacco (Nicotiana tabacum L.) revealed high structural similarity to PAL genes of other plant species. The NtPAL4 gene exists as a single copy in the tobacco plant, and its transcripts were strongly expressed in flowers and leaves. NtPAL4 expression was significantly induced in response to NaCl, mannitol, and cold treatments, but it was not induced by abscisic acid (ABA). NtPAL4 expression decreased gradually after treatment with ABA and $H_2O_2$; however, NtPAL4 transcripts accumulated after treatment with methyl viologen (MV). Our results suggest that the NtPAL4 gene may function in response to abiotic stresses.

• Applied Biological Chemistry
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• 제42권1호
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• pp.34-38
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• 1999

토마토의 genome내에는 적어도 5개 이상의 PAL유전자 좌가 존재한다는 사실을 이 등(1992)이 이미 genomic Southern blot hybridization으로 확인하여 보고하였다. 그러나 본 연구에서 제작한 genomic DNA libraries를 대상으로 검색한 결과 기존에 보고된 PAL유전자 이외에 약 15 kb 와 10 kb에 해당하는 큰 EcoRI 단편을 확보 할 수 있었다. 이들 단편을 BamHI, HindIII등 9종의 제한효소를 사용하여 Southern blot hybridization을 행한 결과 PAL X1의 경우는 모든 효소에 대하여 2개의 단편이 hybridization 되었으며, 특히 BamHI으로 절단하여 얻은 3개의 단편중 두 개는 PAL5 유전자의 exon 2 부위에서 취한 oligomer(18 mer)와 primer extension 반응이 진행되어서 약 200 bp의 PAL유전자와 아주 높은 상동성을 갖는 염기서열이 확인되었다. 따라서 PAL X1유전자는 2 copy의 유전자가 나란히 존재하거나 아니면 염색체 재배열이 진행된 것으로 추정된다. 이러한 결과는 적어도 7개 이상의 PAL유전자 좌가 토마토 염색체내에 존재하는 것으로 사료된다.

• Food Science and Biotechnology
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• 제18권2호
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• pp.494-499
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• 2009

To improve phenylalanine ammonia lyase (E.C.4.3.1.5-PAL) activity in recombinant Escherichia coli, Some approaches for improving phenylalanine ammonia lyase (PAL) activity in recombinant E. coli were developed following preliminary studies by means of response surface method. The results shown that permeabilization with combination of Triton X-100, cetyl trimethyl ammonium bromide (CTAB), and acetone enriched cellular recombinant PAL activity significantly, which improved over 10-fold as compared with the control (untreat cell), as high as 181.37 U/g. The optimum values for the tested variables were Triton X-100 0.108 g/L, CTAB 0.15 g/L, and acetone 45.2%(v/v). Furthermore, a second-order model equation was suggested and then validated experimentally. It was indicated that addition of surfactants and organic solvents made the cells more permeable and therefore allowed easier access of the substrate to the enzyme and excretion of the product, which increased the rate of transport of L-phenylalanine and trans-cinnamic acids. These improved methods of PAL activity enrichment could serve as a rich enzyme source, especially in the biosynthesis of L-phenylalanine.