• The Korean Journal of Food And Nutrition
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• v.9 no.4
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• pp.430-433
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• 1996

The activity of PAL, an initial and key regulatory enzyme in biosynthesis of phenolics, peaked during ripening stage. The enzyme activities were 5.60 and 4.25 units/100g fr. wt. in ripening and overripening stages. The increase in PAL activity in ripening strawberry fruits is due to de novo synthesis of the enzyme. PAL extracts from ripe strawberry fruits have a native molecular weight of about 260, 000 Da.

• Food Science and Biotechnology
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• v.18 no.2
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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.

• Preventive Nutrition and Food Science
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• v.2 no.4
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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.

• 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.

• Food Science and Biotechnology
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• v.18 no.4
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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.

• Plant Biotechnology Reports
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• v.5 no.4
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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.

• Current Research on Agriculture and Life Sciences
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• v.20
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• pp.65-70
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• 2002

This study was conducted to investigate allelopathic potential of various crosses of rice cultivars using their inhibitory effect on barnyardgrass growth under field conditions, and to determine PAL activity and phenolic compounds involved in phenylpropanoid pathway from selected crosses of rice cultivars. Under field conditions, Kouketsumochi, Woo co chin yu possessed higher allelopathic potential inhibiting over 90% of barnyardgrass growth. Crosses of Kouketsumochi/Woo co chin yu, Dongjinbeyo/Kouketsumochi, Dongjinbeyo/Woo co chin yu showed over 80% inhibitory effects on barnyardgrass growth. The highest PAL activity, $63.46{\mu}kats/kg$ proteins was detected in Kouketsumochi which is the most important enzyme in phenylpropanoid pathway and also higher PAL activity in crosses with Kouketsumochi. Content of cinnamate was $2.64{\mu}g/g$ f.w. in Kouketsumochi which was 2 to 5 times higher than other rice cultivars tested, indicating that higher PAL activity resulted in more cinnamate. The similar trends in cinnamate content and PAL activity were observed in crosses of rice cultivars with Kouketsumochi.

• Journal of Life Science
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• v.15 no.2 s.69
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• pp.298-303
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• 2005

This experiment was carried to investigate the optimum time and times of abscisic acid (ABA) treatment for the coloration in 'Kyoho' grapes. The L-phenylalanine ammonia-lyase (PAL) enzyme activities were showed highly in both ABA treatments at veraison and 10 days after veraison. However, PAL enzyme was slightly higher in ABA treatment at 10 days after veraison than in that at veraison. Anthocyanin content showed a tendency that were increased during fruit development after veraison in all treatments, and was the highest in ABA treatment at 10 days after veraison. Fructose and glucose as soluble sugars were detected by HPLC and showed little differences in all treatments. In times of ABA treatment, PAL activity showed a tendency that decreased after increased in all treatments. PAL activity in 2 times treatment of ABA was higher than other treatments. Also, anthocyanin content was highest in 2 times treatment of ABA, as 5 folds of control and folds of 1 time treatment of ABA. Both fructose and glucose contents in all ABA treatments was slightly higher than control.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.7
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• pp.1006-1012
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• 2017

Objective: This study was conducted to investigate effects of zinc (Zn) bearing palygorskite (ZnPal) supplementation on growth performance, hepatic mineral content, and antioxidant status of broilers at early age. Methods: A total of 240 1-day-old Arbor Acres broiler chicks were allocated into 5 treatments with 6 replicates of 8 chicks each. Birds in 5 treatments were fed a basal diet supplemented with 0 (Control group; Analyzed Zn content: 81 mg/kg), 20, 40, 60, and 80 mg/kg Zn as ZnPal for 21 days, respectively. Blood, liver and intestinal mucosa were collected at 21 days of age. Results: Treatments did not affect growth performance of broilers during the 21-day study (p>0.05). The contents of hepatic Zn and magnesium (Mg) were linearly increased (p<0.001) by ZnPal supplementation. ZnPal inclusion linearly (p = 0.007) reduced malondialdehyde (MDA) concentration in serum. The activity of total superoxide dismutase (T-SOD) in liver increased linearly (p = 0.001) with concentration of ZnPal in diet. ZnPal inclusion linearly (p = 0.036) and quadratically (p = 0.005) increased T-SOD activity, and linearly (p = 0.012) increased copper/zinc superoxide dismutase (Cu/Zn SOD) activity in jejunal mucosa. The maximum responses of hepatic and jejunal antioxidant enzymes activities (T-SOD and Cu/Zn SOD) were found when supplementing the basal diet with 60 mg/kg Zn as ZnPal. Furthermore, ZnPal supplementation quadratically (p = 0.001) increased Cu/Zn SOD activity in ileal mucosa, and its maximum activity was observed in the diet supplemented with 20 mg/kg Zn as ZnPal. Conclusion: ZnPal supplementation did not alter growth performance of broilers. Dietary ZnPal inclusion could increase concentrations of hepatic trace minerals (Zn and Mg) and inhibit lipid peroxidation by reducing serum MDA accumulation, with the optimal dosage of Zn from ZnPal being 80 mg/kg diet (analyzed Zn content in the diet: 165 mg/kg), and 60 mg/kg Zn as ZnPal (analyzed Zn content in the diet: 148 mg/kg) was the optimum dosage for broilers to achieve maximum antioxidant enzyme activities.

• Journal of Microbiology and Biotechnology
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• v.30 no.4
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• pp.622-632
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• 2020

Phenylalanine ammonia-lyase (PAL) catalyzes the reversible deamination of phenylalanine to cinnamic acid and ammonia. Algae have been considered as biofactories for PAL production, however, biochemical characterization of PAL and its potency for myristicin biotransformation into MMDA (3-methoxy-4, 5-methylenedioxyamphetamine) has not been studied yet. Thus, PAL from Anabaena flos-aquae and Spirulina platensis has been purified, comparatively characterized and its affinity to transform myristicin was assessed. The specific activity of purified PAL from S. platensis (73.9 μmol/mg/min) and A. flos-aquae (30.5 μmol/mg/min) was increased by about 2.9 and 2.4 folds by gel-filtration comparing to their corresponding crude enzymes. Under denaturing-PAGE, a single proteineous band with a molecular mass of 64 kDa appeared for A. flos-aquae and S. platensis PAL. The biochemical properties of the purified PAL from both algal isolates were determined comparatively. The optimum temperature of S. platensis and A. flos-aquae PAL for forward or reverse activity was reported at 30℃, while the optimum pH for PAL enzyme isolated from A. flos-aquae was 8.9 for forward and reverse activities, and S. platensis PAL had maximum activities at pH 8.9 and 8 for forward and reverse reactions, respectively. Luckily, the purified PALs have the affinity to hydroaminate the myristicin to MMDA successfully in one step. Furthermore, a successful method for synthesis of MMDA from myristicin in two steps was also established. Gas chromatography-mass spectrometry (GC-MS) analysis was conducted to track the product formation.