• Title/Summary/Keyword: Phenylalanine ammonia-lyase

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Biosynthesis of Pinocembrin from Glucose Using Engineered Escherichia coli

  • Kim, Bong Gyu;Lee, Hyejin;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1536-1541
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    • 2014
  • Pinocembrin is a flavonoid that exhibits diverse biological properties. Although the major source of pinocembrin is propolis, it can be synthesized biologically using microorganisms such as Escherichia coli, which has been used to synthesize diverse natural compounds. Pinocembrin is synthesized from phenylalanine by the action of three enzymes; phenylalanine ammonia lyase (PAL), 4-coumarate:CoA ligase (4CL), and chalcone synthase (CHS). In order to synthesize pinocembrin from glucose in Escherichia coli, the PAL, 4CL, and CHS genes from three different plants were introduced into an E. coli strain. Next, we tested the different constructs containing 4CL and CHS. In addition, the malonyl-CoA level was increased by overexpressing acetyl-CoA carboxylase. Through these strategies, a high production yield (97 mg/l) of pinocembrin was achieved.

INDUCTION OF SYSTEMIC RESISTANCE IN CUCUMBER AGAINST ANTHRACNOSE BY PLANT GROWTH PROMOTING FUNGI

  • Hyakumachi, Mitsuro
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1997.06a
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    • pp.47-55
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    • 1997
  • Plant growth promoting fungi(PGPF) obtained from zoysiagrass rhizosphere offer dual advantages - induse systemic disease resistance response in cucumber to C. orbiculare infection and cause enhancement of plant growth and increase yield. PGPF protected plants either by colonizing roots or by their metabolites. PGPF offer an advantage by protecting plants for more than 9 weeks and 6 week in the greenhouse and field. PGPF-induced plants limited pathogen spore germination and decreased the number of infection hyphae on the leaf, and increased lignification at places of attempted pathogen infection, thus reducing the pathogen spread. PGPF elicited increased activities of chitinascs, glucanases, peroxidase, polyphenol oxidase, and phenylalanine ammonia lyase to C. orbiculare infection in cucumber plants. The role of PGPF in elevating cucumber defense response to pathogen infection suggests potential application of PGPF as biological control agents.

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Enzyme Activities and Compounds Related to Self-Defense in UV-Challenged Leaves of Rice

  • Kim, Hak-Yoon;Shin, Dong-Hyun;Sohn, Dae-Seung;Lee, In-Jung;Kim, Kil-Ung;Lee, Sang-Chul;Jeong, Hyung-Jin;Cho, Moon-Soo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.1
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    • pp.22-28
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    • 2001
  • The induction of enzymes and the accumulation of their end products associated with self-defense mechanism in rice were investigated. When rice leaves were irradiated with UV light, activities of diterpene cyclase, phenylalanine ammonia-lyase (PAL), and cinnamic acid 4-hydroxylase (CA4H) were induced and rice phytoalexin, momilactone A was accumulated. The content of p-coumaric acid in rice leaves was closely correlated with self-defense or allelopathic potential against barnyardgrass. UV-challenged rice leaves gave rise to the inhibition of barnyardgrass growth.

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Microbial Aspartase and Its Activity on Deamination of L-Aspartyl-L-Phenylalanine Methyl Eester

  • Chang, Wonyoon;Goo, Yang-Mo
    • Archives of Pharmacal Research
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    • v.11 no.2
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    • pp.139-144
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    • 1988
  • Examination of many microorganisms and soil isolated for the activity of aspartase proved that R, rubra, G, suboxydans, A. versicolor, P. purpurogenum, E. coli, Ps. aeruginosa, A. gigantus, A, unguis, A. parasiticus and a soil isolate (S-90) had high activity of aspartase. Comparison of the activity of the aspartase by cell free extracts of these microorganisms with the activity of the enzyme catalyzing the deamination of aspartame by the same cell free extracts showed similar kinetic characteristics. The aspartase existing in the cell free extracts seemed to catalyze the deamination of aspartame, too.

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Effects of Abscisic acid and Temperature on the Anthocyanin Accumulation in Seedlings of Arabidopsis thaliana

  • Song Ju-Yeun;Kim Tae-Yun;Hong Jung-Hee
    • Journal of Environmental Science International
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    • v.14 no.12
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    • pp.1093-1102
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    • 2005
  • Effects of abscisic acid(ABA) and temperature on the anthocyanin accumulation and phenylalanine ammonia Iyase(PAL) activity were investigated in seedlings of Arabidopsis thaliana. In time course study, exogenous application of ABA $(50-1000\;{\mu}M)$ led to a noticeable increase in anthocyanin pigments which persisted over the following 5 days. Anthocyanins increased in concert with the chlorophyll loss. The activity of PAL, a key enzyme in the phenylpropanoid pathway, increased on exposure to ABA and reached maximum on the 4th day, This result shows that anthocyanin synthesis and PAL activity have a close physiological relationships. In the effects of temperatures ($10^{\circ}C,\;17^{\circ}C,\;25^{\circ}C$and $30^{\circ}C$) on anthocyanin accumulation and PAL activity in seedlings, a moderate-low temperatures ($17^{\circ}C$) enhanced both anthocyanin content and PAL activity, whereas elevated temperatures ($30^{\circ}C$) showed low levels of anthocyanin and PAL activity, suggesting a correlation between temperature-induced anthocyanin synthesis and the accumulation of PAL mRNA. Simultaneous application of ABA with temperatures Induced higher anthocyanin synthesis and PAL activity in seedlings than ABA or temperature stress alone. Moderate-low temperature with ABA exposure elicited the maximal induction of anthocyanin synthesis and PAL activity. Therefore, ABA treatment significantly increased thermotolerance in .A. thalinan seedlings. Ethephon and ABA showed similar mode of action in physiological effects on anthocyanin accumulation and PAL activity. Our data support that anthocyanins may be protective in preventing damage caused by environmental stresses and play an important role in the acquisition of freezing tolerance.

Phenylketonuria: Current Treatments and Future Developments (페닐케톤뇨증의 치료: 현재와 미래)

  • Lee, Jeongho
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.20 no.2
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    • pp.37-43
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    • 2020
  • Phenylketonuria is the most prevalent disorder caused by an inborn error in aminoacid metabolism. It results from mutations in the phenylalanine hydroxylase (PAH) gene. If untreated or late treated, results in profound and irreversible mental disability. Newborn screening test identify patients with phenylketouria. The early initiation of a phenylalanine restricted diet very soon prevents most of the neuropsychiatric complications. However, the diet therapy is difficult to maintain and compliance is poor, especially in adolescents and adulthood. Since 2015, American Medical College of Medical Genetics and Genomics (ACMG) recommended more strong restrictive diet therapy for target blood level of phenylalanine (<360 umol/L). For over four decades the only treatment was a very restrictive low phenylalanine diet. This changed in 2007 with the approval of cofactor therapy (Tetrahydrobiopterin, BH4) which is effective in up to 30% of patients. Data from controlled clinical trials with sapropterin dihydrochloride indicate a similar occurrence of all-cause adverse events with this treatment and placebo. Large neutral aminoacids (LNAA) competes with phenylalanine for transport across the blood-brain-barrier and have a beneficial effect on executive functioning. A new therapy has just been approved that can be effective in most patients with PAH deficiency regardless of their degree of enzyme deficiency or the severity of their phenotype. Phenylalanine ammonia lyase (PAL-PEG) was approved in the USA by FDA in May of 2018 for adult patients with uncontrolled blood phenylalanine concentrations on current treatment. Nucleic acid therapy (therapeutic mRNA or gene therapy) is likely to provide longer term solutions with few side effects.

Expression of Genes Affecting Skin Coloration and Sugar Accumulation in 'Hongro' Apple Fruits at Ripening Stages in High Temperatures (고온에 의한 변색단계별 '홍로' 사과의 착색 및 당 축적 관련 유전자 발현 분석)

  • Kim, Seon Ae;Ahn, Soon Young;Yun, Hae Keun
    • Journal of Bio-Environment Control
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    • v.25 no.1
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    • pp.9-15
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    • 2016
  • High temperature is one of the important environmental factors limiting cultivation of apple (Malus domestica Borkh). The expression of genes related with anthocyanin synthesis and sugar accumulation in response to high temperature was studied in the 'Hongro' apple fruits at different developmental stages in different temperature conditions through real-time PCR. Expression of ${\hat{a}}$-amylase (BMY) and polygalacturonase (PG) genes related with sugar synthesis was higher in late ripening stages than in initial ripening stages. Expression of four genes such as phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), flavanone 3-hydroxylase (F3H), and malate dehydrogenase (MDH), which were related with fruit skin coloration, increased gradually in apple fruits of the middle and late ripening stages. Interestingly, the expressions of all genes were highly inhibited expressed at $30-35^{\circ}C$ compared to $25^{\circ}C$ in all ripening stages. In the further work, investigation of expression levels of various genes could be conducted in the level of transcriptomics in fruits at the middle ripening stages to get meaningful information of ripening metabolism in apple in high temperatures.

Introduction of Shiva Gene into tobacco and Potato Using Tissue-Specific Tomato PAL Promoter (조직특이성 promoter를 이용한 Shiva 유전자의 식물체내 도입)

  • 이정윤;이신우;박권우
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.2
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    • pp.109-113
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    • 1998
  • In this study we tried to transform an antimicrobial peptide gene (Shiva) under the promoter of tomato phenylalanine ammonia-lyase (tPAL5) into tobacco and potato plants. Antimicrobial peptide gene was isolated originally from giant silk moth (Hyalophora cecropia) and modified ie nucleotide sequence to increase antimicrobial activity. Transgenic tobacco plants were regenerated and their seeds were tested on the media containing kanamycin (500 mg/L). The results of PCR amplification and genomic Southern blot hybridization confirmed the integration of construct (tPAL5 promoter-Shiva-NOS-GUS-NOS) into chromosome. We observed that one of the transgenic tobacco plants showed chromosome rearrangement when integrated. In case of potato transformation, the efficiency of regeneration was maximized at the medium containing Zeatin 2mg/L, NAA 0.01mg/L, GA$_3$ 0.1mg/L. We also observed the high expression of GUS (${\beta}$-glucuronidase) enzyme which was located next to the terminator sequence of nopaline synthase gene (NOS) in the vascular tissue of stem, leaves of transgenic potatoes. This result suggested that a short sequence of Shiva gene (120 bp) and NOS terminator sequence might be served as a leader sequence of transcript when translated.

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Induction of Defense Related Enzymes and Pathogenesis Related Proteins in Pseudomonas fluorescens-Treated Chickpea in Response to Infection by Fusarium oxysporum f. sp. ciceri

  • Saikia, Ratul;Kumar, Rakesh;Singh, Tanuja;Srivastava, Alok K.;Arora, Dilip K.;Lee, Min-Woong
    • Mycobiology
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    • v.32 no.1
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    • pp.47-53
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    • 2004
  • Pseudomonas fluorescens 1-94 induced systemic resistance in chickpea against Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceri by the synthesis and accumulation of phenolic compounds, phenylalanine ammonia lyase(PAL) and pathogenesis related(PR) proteins(chitinase, $\beta$-1,3-glucanase and peroxidase). Time-course accumulation of these enzymes in chickpea plants inoculated with P. fluorescens was significantly(LSD, P=0.05) higher than control. Maximum activities of PR-proteins were recorded at 3 days after inoculation in all induced plants; thereafter, the activity decreased progressively. Five PR peroxidases detected in induced chickpea plants. Molecular mass of these purified peroxidases was 20, 29, 43, 66 and 97 kDa. Purified peroxidases showed antifungal activity against plant pathogenic fungi.

Role of Riboflavin in Induced Resistance against Fusarium Wilt and Charcoal Rot Diseases of Chickpea

  • Saikia Ratul;Yadav Mukesh;Varghese Saju;Singh Bhim Pratap;Gogoi Dip K;Kumar Rakesh;Arora Dilip K
    • The Plant Pathology Journal
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    • v.22 no.4
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    • pp.339-347
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    • 2006
  • Riboflavin caused induction of systemic resistance in chickpea against Fusarium wilt and charcoal rot diseases. The dose effect of 0.01 to 20 mM riboflavin showed that 1.0 mM concentration was sufficient for maximum induction of resistance; higher concentration did not increase the effect. At this concentration, riboflavin neither caused cell death of the host plant nor directly affected the pathogen's growth. In time course observation, it was observed that riboflavin treated chickpea plants were inducing resistance 2 days after treatment and reached its maximum level from 5 to 7 days and then decreased. Riboflavin had no effect on salicylic acid(SA) levels in chickpea, however, riboflavin induced plants found accumulation of phenols and a greater activities of phenylalanine ammonia lyase(PAL) and pathogenesis related(PR) protein, peroxidase was observed in induced plant than the control. Riboflavin pre-treated plants challenged with the pathogens exhibited maximum activity of the peroxidases 4 days after treatment. Molecular weight of the purified peroxidase was 42 kDa. From these studies we demonstrated that riboflavin induced resistance is PR-protein mediated but is independent of salicylic acid.