• 한국식품영양과학회지
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• 제16권1호
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• pp.10-17
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• 1987

고추의 배양세포(培養細胞)에 있어서 고추의 신미성분(辛味成分)인 capsaicinoids 생성(生成)에 미치는 growth regulator의 영향을 조사하기 위하며 growth regulator들을 각각 함유한 Linsmaier-Skoog 기본배지에서 배양(培養)한 callus의 capsaicinoids을 GLC에 의하여 조사하고 또한 capsaicinoids의 중간대사 산물인 phenylpropanoids와 phenylalanine ammonia-lyase activity을 측정하여 다음과 같은 결과를 얻었다. 1. Growth regulators가 배양세포(培養細胞)의 capsaicinoids 생성(生成)에 미치는 영향을 조사한 결과 growth regulators을 달리한 배지(培地)에서 생성(生成)된 capsaicinoids의 함량(含量)은 $2,4-D\;10^{-6}M$에서 가장 capsaicinoids의 생성량(生成量)이 많았으며 $IAA\;10^{-6}M,\;NAA\;10^{-6}M,\;kinetin\;10^{-6}M$의 배지(培地)에서는 2,4-D에 비(比)하여 각각 65%, 38%, 68%이었다. 2. Growth regulators을 달리한 기본배지에서 생성(生成)된 phenylpropanoids는 cinnamic acid, coumaric acid의 생성(生成)은 5, 15, 25일(日)에 점진적으로 증가하였으며 NAA 함유 배지에서는 2,4-D에 비해 약 30%, kinetin, IAA 함유배지에서는 약 70% 생성(生成)되었다. 3. $2,4-B\;10^{-6}M$을 함유한 배양세포(培養細胞)에서 phenylalanine ammonia-lyase activity을 측정한 결과 배양후(培養後) 5, 15, 25일(日) 배양일수에 따라 phenylalanine ammonia-lyase activity는 신선중량 g당 생성된 cinnamate는 각각 52, 81 및 209 % nmole로 증가하였다.

• BMB Reports
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• 제32권2호
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• pp.203-209
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• 1999

Three-week grown Arabidopsis thaliana leaves were wounded by cutting whole leaves with a razor blade into pieces (about$3\;mm\;{\times}\;3\;mm$) submerged in various solutions, and incubated in a growth chamber for 24 h. We measured and compared activities of several enzymes such as phenylalanine ammonia-lyase (PAL), tyrosine ammonia-lyase (TAL), thioredoxin, thioredoxin reductase, thioltransferase, glutathione reductase, and $NADP^+$ -malate dehydrogenase. PAL activity was decreased in $HgCl_2$-, $CdCl_2$-, and glyphosate-treated leaf slices, and could not be detected after treatment with $CdCl_2$. TAL activity was found to be maximal in the $CdCl_2$-treated leaf slices. Activity of thioredoxin, a small protein known as a cofactor of ribonucleotide reductase and a regulator of photosynthesis, was significantly increased in the $CdCl_2$-treated leaf slices, while thioredoxin reductase activity was maximal in the $HgCl_2$-treated leaf slices. Thioltransferase and glutathione reductase activities were significantly decreased in the $HgCl_2$-treated leaf slices. $NADP^+$ -malate dehydrogenase activity remained relatively constant after the chemical treatments. Our results strongly indicate that sulfhydryl-related and phenylpropanoid-synthesizing enzyme activities are affected by chemical treatments such as hydrogen peroxide, heavy metals, and glyphosate.

• Journal of Microbiology and Biotechnology
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• 제6권5호
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• pp.347-351
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• 1996

Flavonoid production by suspended cells of Scutellaria baicalensis Georgi was studied and the medium was optimized for cell growth and baicalin production. In SH medium the flavonoid production was not closely associated with the cell growth. A modified SH medium, FPM, was therefore designed for enhanced baicalin production. In FPM, both cell growth and baicalin production were increased by 1.5 times and 1.67 times than in the original SH medium, respectively. The increases could be attributed to the increased metabolic activities involved in the flavonoid biosynthesis as represented by enhanced activities of phenylalanine ammonia-lyase.

• Journal of Plant Biology
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• 제37권3호
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• pp.263-269
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• 1994

Involvement of ethylene and Ca2+ on the induction of phenylalanine ammonia-lyase (PAL) was investigated in Daucus carota L. suspension culture system. Ethylene production started to increase about 3 h after glucan treatment. And the maximal induction of ethylene was preceded by PAL induction by 30 min. After the treatment of ethrel, PAL activity was increased. When cells were treated with glucan and Co2+, PAL activity was simultaneously reduced. Ethylene production was reduced dramatically in calcium-free medium, even though glucan was treated. PAL activity and ethylene producton was inhibited conspicuously when ethylene glycolbis($\beta$-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA) was treated with glucan. Verapamil and trifluoperazine also inhibited PAL activity. When cells were treated with calcium ionophore A23187, PAL activity was increased in nontreated medium. We report here PAl activity is increased in related to ethylene production and involvement of Ca2+ in glucan-treated carrot suspension cells.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2007년도 제48회 학술심포지움 및 추계국제학술대회
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• pp.99.2-99.2
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• 2007

See Full Text

• 한국약용작물학회:학술대회논문집
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• 한국약용작물학회 2010년도 심포지엄 및 추계학술발표회
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• pp.390-391
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• 2010

• 생명과학회지
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• 제19권9호
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• pp.1226-1231
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• 2009

페닐케톤뇨증은 상염색체 열성으로 유전되며, phenylalanine-4-hydroxylase (PAH, EC 1.14.16.1)의 돌연변이에 의해 효소 불활성화를 초래하는 질환이다. 최근 유전자 재조합된 phenylalanine ammonia-lyse (PAL)에 의한 효소 대체요법이 보고된 바 있다. 이 효소를 경구용 약제로 개발하기 위하여 효소활성을 나타내기 위한 최적 조건들을 알아야 하며, 위장관내 소화효소에 의해 분해되지 않는 구조적 안정성을 유지하여야 한다. 따라서 본 연구에서는 PAL의 생화학적 특성을 규명하고, 이를 바탕으로 위장관내 소화효소로부터 저항할 수 있는 변이형들을 만들고자 하였으며, 이러한 구조적 변화를 통하여 효소의 특이 활성도가 유지될 수 있는지를 보고자 하였다. PAL의 특이 활성도를 측정하였고, 효소 활성을 나타내기 위한 최적 pH, 온도 변화에 따른 효소 활성도, 단백분해효소에 의한 활성도 변화를 측정하였다. PAL의 Vmax는 페닐알라닌과 티로신에 대하여 각각 1.77, $0.47{\mu}mol$/ mg x protein로 나타났으며, Km은 페닐알라닌에 대하여 $4.77{\times}10^{-4}\;M$,티로신에 대하여 $4.37{\times}10^{-4}\;M$로 나타났다. 또한 pH 8.5에서 가장 높은 활성을 나타내었는데, 이는 소장의 평균 pH와 유사하다. PAL의 효소 활성은 $-80^{\circ}C$에서 5개월 동안 유지되었으며, $4^{\circ}C$에서 1주일 동안 93.4%의 활성을 유지하였다. PAL은 키모트립신에 의해 쉽게 분해되었으며, 이보다 약한 정도로 트립신, elastase, carboxypeptidase A, B에 의해 분해 되었다. 췌장 소화효소에 대한 저항성을 증가시키기 위하여 트립신, 키모트립신 절단부위 아미노산을 변이시켜 유전자 변이형을 만들었고, 효소 활성도를 측정하였다. 6개의 유전자 변이형은 모두 저하된 효소 활성도를 나타내었는데, Y110H는 0.084, Y110A와 Y110L은 0, R123A는 0.11, R123H는 0.074, R123Q는 0.033으로 나타났다. 이러한 결과는 트립신 및 키모트립신 절단부위 아미노산이 PAL의 효소 활성에 필수적인 역할을 하고 있음을 나타낸다. PAL 변이형은 단백분해작용으로부터 보호할 수 있는 전처치 방법이지만, 페닐알라닌을 효과적으로 저하시키기 위해서 효소활성을 유지할 수 있는 다음 단계의 처치가 필요하다.

• The Plant Pathology Journal
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• 제24권2호
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• pp.171-177
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• 2008

In this study, we investigated the induced defense response and protective effects against Stemphylium vesicarium by application of benzothiadiazole ($Bion^{(R)}$) and di-potassium phosphate salt $(K_2HPO_4)$ to onion. Onion leaves were sprayed with $Bion^{(R)}$ and $K_2HPO_4$, then inoculated 2 days later with a virulent strain of S. vesicarium under greenhouse conditions. Disease severity and activities of peroxidase (PO), polyphenoloxidase, phenylalanine ammonia-lyase (PAL) and phenol contents were evaluated in the treated leaf tissues. Reduction in the disease severity was observed in plants treated with $Bion^{(R)}$ and $K_2HPO_4$. Onion plants treated with $Bion^{(R)}$ and $K_2HPO_4$ and inoculated with the pathogen showed significantly higher PAL activity, PO activity, and phenol contents than inoculated water-treated plants 2 days after the treatment. In conclusion, the results of this study provide evidence that application of simple non-toxic chemical solutions as di-potassium phosphate and $Bion^{(R)}$ can control Stemphylium leaf blight of onion.

• 식물조직배양학회지
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• 제25권2호
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• pp.89-93
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• 1998

Tomato phenylalanine ammonia-lyase 5 (tPAL5) was identified that alternate initiation sites were utilized differentially in response to environmental stimuli (Lee et al, 1992b). In this study, we tried to look into tissue -or cell- specific expression pattern of tPAL5 gene by fusing with ${\beta}-glucuronidase$ (GUS) gene in transgenic tobacco plants. In transgenic plants, root and stem extracts contained 8~12 fold higher levels of GUS activity than petiole or leaf tissue while the highest levels of induction was observed from leaf tissue by mechanical wounding (5~11 fold). In trans-sections of stems and petioles, GUS activity was restricted to phloem cells(outer region) of developing vascular bundle and mainly at apical tip region in the root tissues. The levels of GUS activity was drastically reduced (10~12 fold reduction) when the 5'-upstream region of tPAL5 gene (-1151bp from ATG codon) was deleted up to -665. The levels of GUS expression, however, raised up by 6~8 fold when deleted up to -455. Therefore, we conclude that there are positive cis-elements at the region -1151 to -1008 and at -455 to -195 while the negative cis-element is at -1008 to -455.

• Plant Biotechnology Reports
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• 제5권4호
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• pp.301-308
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• 2011

This study tested the relative and combined efficacy of ShPx2 and ShPAL transgenes by comparing Nicotiana tabacum hybrids with enhanced levels of $_L$-phenylalanine ammonia lyase (PAL) activity and cationic peroxidase (Prx) activity with transgenic parental lines that overexpress either transgene. The PAL/Prx hybrids expressed both transgenes driven by the 35S CaMV promoter, and leaf PAL and Prx enzyme activities were similar to those of the relevant transgenic parent and seven- to tenfold higher than nontransgenic controls. Lignin levels in the PAL/Prx hybrids were higher than the PAL parent and nontransgenic controls, but not significantly higher than the Prx parent. All transgenic plants showed increased resistance to the necrotrophs Phytophthora parasitica pv. nicotianae and Cercospora nicotianae compared to nontransgenic controls, with a preponderance of smaller lesion categories produced in Prx-expressing lines. However, the PAL/Prx hybrids showed no significant increase in resistance to either pathogen relative to the Prx parental line. These data indicate that, in tobacco, the PAL and Prx transgenes do not act additively in disease resistance. Stacking with Prx did not prevent a visible growth inhibition from PAL overexpression. Practical use of ShPAL will likely require more sophisticated developmental control, and we conclude that ShPx2 is a preferred candidate for development as a resistance transgene.