Molecular characterization of a phenylalanine ammonia-lyase gene (BoPAL1) from Bambusa oldhamii

Abstract

Phenylalanine ammonia-lyase is the first enzyme of general phenylpropanoid pathway. A PAL gene, designated as BoPAL1, was cloned from a Bambusa oldhamii cDNA library. The open reading frame of BoPAL1 was 2,139 bp in size and predicted to encode a 712-amino acid polypeptide. BoPAL1 was the first intronless PAL gene found in angiosperm plant. Several putative cis-acting elements such as P box, GT-1motif, and SOLIPs involved in light responsiveness were found in the 5'-flanking sequence of BoPAL1 which was obtained by TAIL-PCR method. Recombinant BoPAL1 protein expressed in Pichia pastoris was active. The optimum temperature and pH for BoPAL1 activity was 50°C and 9.0, respectively. The molecular mass of recombinant BoPAL1 was estimated as 323 kDa using gel filtration chromatography and the molecular mass of full-length BoPAL was about 80 kDa, indicating that BoPAL1 presents as a homotetramer. The Km and kcat values of BoPAL1 for L-Phe were 1.01 mM and 10.11 s(-1), respectively. The recombinant protein had similar biochemical properties with PALs reported in other plants.

Characterization and primary functional analysis of phenylalanine ammonia-lyase gene from Phyllostachys edulis

“…Using the Michaelis-Menten equation, hyperbolic plot was obtained by triplicate reactions and the averages were converted to double reciprocal plot (Lineweaver-Burk plot) to calculate the K m (Michaelis-Menten constant) and V max (Hsieh et al 2011). TAL activity of PePAL was assayed by monitoring p-coumaric acid formation at 310 nm (Kyndt et al 2002).…”

“…The annual forest production of P. edulis shoot and timber in China was about US $5 billion (Jiang 2002). When bamboo shoots are harvested, PAL activity might be induced to play a role in the biosynthesis of phenolic compounds, which causes hardening and browning substance deposition in the shoots and in the development of a bitter taste (Hsieh et al 2011). Meanwhile, PAL is also one of the key enzymes of lignin synthesis which affects the properties of bamboo timber.…”

Biochemical Evaluation of Phenylalanine Ammonia Lyase (PAL) from Endemic Cyathobasis fruticulosa (Bunge) Aellen. for the Dietary Treatment of Phenylketonuria (PKU)

“…The PAL activity of Cyathobasis fruticulosa is reportedly higher ((64.9±0.1) U/mg) than those of other plants, including Bambusa oldhamii with a PAL activity of 0.19 U/mg (25), Fragaria × ananassa 44.5 U/mg (28), Cucurbita pepo 26.6 U/mg (9), and Glycine max 0.43 U/mg (29). Plants are able to utilize light, carbon dioxide and various inorganic substances in an effi cient manner for their relatively simple vital demands.…”

“…The membrane was rinsed twice with double distilled H 2 O for 2 min and dried. The chromogenic band was visualized with UVP BioSpectrum 410 (Upland, CA, USA) imaging system (24,25).…”

Molecular cloning, expression, and subcellular localization of a PAL gene from Citrus reticulata under iron deficiency

“…In this paper, real time qPCR analysis reveals that a high CrPAL1 expression was found in roots (Table 2). Expression of the PAL gene in roots could be induced by different abiotic stimulation, such as low content of nitrogen, phosphate and iron, or by signalling compounds such as methyl jasmonate, jasmonic acid, H 2 O 2 , and salicylic acid (Hsieh et al 2011). Studies have also discovered that the total amount of phenolic compounds was higher under Fe deficiency (Hell and Stephan 2003).…”