• Journal of the Korean Society of Food Science and Nutrition
• /
• v.16 no.1
• /
• pp.10-17
• /
• 1987

In order to investigate the effects of growth regulators on the formation of capsaicinoids in callus of Capsicum annuum L. tissues were cultured in the Linsmaier and Skoog RM 1964 medium containing various growth regulators. Production of capsaicinoids during culture was monitored by gas chromatography. In the presence of $10^{-6}M$ of 2,4-D and kinetin in the medium, $1182{\mu}g$ of capsaicinoids were formed per 100g dry wt. of tissue, of which was greater than with any of three other growth regulators. IAA, NAA, and kinetin of same concentrations had 65%, 38%, 68% effect of 2.4-D in capsaicinoids formation, respectively. Production of capsaicinoids increased gradually in the presence of 2,4-B as culture period was proceeded. Of phenylpropanoids formed, cinnamic acid and coumaric acid were not significantly different in their levels, although growth regulators were varied. On the other hand, caffeic acid and ferulic acid formation were highest in the presence of 2,4-D. Effects of kinetin and IAA were about 70 percent of that of 2,4-D, whereas NAA had only about 30 percent effect. Phenylalanine ammonia-lyase activity in cultured tissue was increased during the periods; 52, 81, and 209 n moles of cinnamic acid per g fresh wt. were formed after 5, 15, and 25 days of culture, respectively.

• BMB Reports
• /
• v.32 no.2
• /
• pp.203-209
• /
• 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
• /
• v.6 no.5
• /
• pp.347-351
• /
• 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
• /
• v.37 no.3
• /
• pp.263-269
• /
• 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.

• Proceedings of the Korean Society of Life Science Conference
• /
• 2007.10a
• /
• pp.99.2-99.2
• /
• 2007

See Full Text

• 한국약용작물학회:학술대회논문집
• /
• 2010.10a
• /
• pp.390-391
• /
• 2010

• Journal of Life Science
• /
• v.19 no.9
• /
• pp.1226-1231
• /
• 2009

Enzyme substitution with recombinant phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is currently being explored for treatment of phenylketonuria (PKU), an autosomal recessive genetic disorder with mutations of the gene encoding phenylalanine-4-hydroxylase (EC 1.14.16.1). However, oral administration of PAL is limited because of proteolytic digestion in the gastrointestinal tract. The aim of this study was to determine the biochemical properties of PAL and delinate the susceptibility of wild-type PAL to pancreatic proteolysis by exploring several mutants, and to develop therapeutic drugs with PAL for PKU. The specific activity of PAL was assayed and its optimal pH, temperature stability, and intestinal protease susceptibility were investigated. Its $V_{max}$ values for phenylalanine and tyrosine were 1.77 and $0.47{\mu}mol$/ min/mg protein, respectively, and its $K_m$ values were $4.77{\times}10^{-4}$ and $4.37{\times}10^{-4}\;M$, respectively. PAL showed an optimal pH at 8.5, corresponding to the average pH range of the small intestine. It showed no loss of activity at $-80^{\circ}C$ for 5 months and possessed 93.4% of its activity under $4^{\circ}C$ for 4 wks. PAL was susceptible to chymotrypsin digestion and, to a lesser extent, to trypsin, elastase, carboxypeptidase A, and B. The trypsin and chymotrypsin cleaving sites were mutated to investigate protection from pancreatic digestion and the specific activities of these mutants were evaluated. The six mutants displayed low specific activities compared to the wild-type, suggesting that the primary trypsin and chymotrypsin cleaving sites may be essential for catalytic reaction. The PAL mutants could therefore be applied as a pretreatment modality without susceptibility to proteolytic attack, however, additional modification for enhancing enzymatic activity is needed to reduce the Phe levels effectively.

• The Plant Pathology Journal
• /
• v.24 no.2
• /
• pp.171-177
• /
• 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.

• Korean Journal of Plant Tissue Culture
• /
• v.25 no.2
• /
• pp.89-93
• /
• 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
• /
• v.5 no.4
• /
• pp.301-308
• /
• 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.