Cloning and expression of a phenylalanine ammonia-lyase gene (BoPAL2) from Bambusa oldhamii in Escherichia coli and Pichia pastoris

Hsieh, Lu‐Sheng; Yeh, Chen-Pin; Pan, Horng‐Bin; Cheng, Chieh-Yang; Yang, Chien‐Chih; Lee, Ping-Du

doi:10.1016/j.pep.2010.01.009

Cited by 36 publications

(31 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Residues shaded in solid black match the consensus exactly, and the conserved active site motif (Ala-Ser-Gly) which can be converted into a MIO prosthetic group indicated by rectangle to L-phenylalanine was calculated as 422 lM and 45.9 nM s -1 , respectively. This K m value was similar to that of rice PAL (500 lM) (Sarma and Sharma 1999), and about half of BoPAL1 (1.01 mM) (Hsieh et al 2010b). The recombinant protein also exhibited activity towards L-tyrosine with K m and V max value as 78 lM and 7.09 nM s -1 , respectively, and the K m value was about two times higher than that of maize PAL (41 lM) (Rösler et al 1997).…”

Section: Purification and Functional Analysis Of Recombinant Pepalsupporting

confidence: 80%

“…The most closely related sequence to PePAL was BoPAL4, with 97.7 % identity of the amino acid sequence (685/701 amino acids). Furthermore, previous research on BoPAL2 and BoPAL4 showed that the recombinant proteins exhibited both PAL and TAL activities (Hsieh et al 2010b). PePAL was highly similar to BoPAL4, which indicated that it might have dual PAL/TAL activities.…”

Section: Phylogenetic Analysis Of Pepalmentioning

confidence: 83%

“…PAL activity was assayed by measuring trans-cinnamic acid formation by the absorbance increase at 290 nm (Hsieh et al 2010b).…”

Section: Expression Of the Recombinant Proteinmentioning

confidence: 99%

See 2 more Smart Citations

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

et al. 2012

View full text Add to dashboard Cite

show abstract

Section: Purification and Functional Analysis Of Recombinant Pepalsupporting

confidence: 80%

Section: Phylogenetic Analysis Of Pepalmentioning

confidence: 83%

See 1 more Smart Citation

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

et al. 2012

View full text Add to dashboard Cite

show abstract

“…Several enzymes have previously been characterized both in vivo and in vitro, and the data are available from both patent and scientific literature (3,(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32). However, the conditions of kinetic analysis are often quite different and may not represent the extent to which the enzymes perform in vivo.…”

mentioning

confidence: 99%

Highly Active and Specific Tyrosine Ammonia-Lyases from Diverse Origins Enable Enhanced Production of Aromatic Compounds in Bacteria and Saccharomyces cerevisiae

Jendresen

Stahlhut

et al. 2015

Appl Environ Microbiol

158

183

View full text Add to dashboard Cite

Phenylalanine and tyrosine ammonia-lyases form cinnamic acid and p-coumaric acid, which are precursors of a wide range of aromatic compounds of biotechnological interest. Lack of highly active and specific tyrosine ammonia-lyases has previously been a limitation in metabolic engineering approaches. We therefore identified 22 sequences in silico using synteny information and aiming for sequence divergence. We performed a comparative in vivo study, expressing the genes intracellularly in bacteria and yeast. When produced heterologously, some enzymes resulted in significantly higher production of p-coumaric acid in several different industrially important production organisms. Three novel enzymes were found to have activity exclusively for phenylalanine, including an enzyme from the low-GC Gram-positive bacterium Brevibacillus laterosporus, a bacterial-type enzyme from the amoeba Dictyostelium discoideum, and a phenylalanine ammonia-lyase from the moss Physcomitrella patens (producing 230 M cinnamic acid per unit of optical density at 600 nm [OD 600 ]) in the medium using Escherichia coli as the heterologous host). Novel tyrosine ammonia-lyases having higher reported substrate specificity than previously characterized enzymes were also identified. Enzymes from Herpetosiphon aurantiacus and Flavobacterium johnsoniae resulted in high production of p-coumaric acid in Escherichia coli (producing 440 M p-coumaric acid OD 600 unit ؊1 in the medium) and in Lactococcus lactis. The enzymes were also efficient in Saccharomyces cerevisiae, where p-coumaric acid accumulation was improved 5-fold over that in strains expressing previously characterized tyrosine ammonia-lyases. Small organic molecules of biotechnological interest include aromatic structures that are derived from p-coumaric acid (pHCA). pHCA can be formed from phenylalanine either through deamination to cinnamic acid (CA) by phenylalanine ammonialyase (PAL; EC 4.3.1.24) and subsequent hydroxylase activity by trans-cinnamate-4-monooxygenase or through deamination of tyrosine by tyrosine ammonia-lyase (TAL; EC 4.3.1.23) (Fig. 1). Considering that the route from phenylalanine requires activity of a P450 enzyme, which has been shown to be the limiting step in previous engineering strategies (1, 2), deamination of tyrosine for production of pHCA may be preferred in microbial cell factories. Tyrosine ammonia-lyases (TAL) have been employed for the production of plant biochemicals and aromatic compounds, e.g., for pHCA itself (3-5), or in the production of stilbenes, such as resveratrol (6, 7), flavonoids, such as naringenin (8, 9), cinnamoyl anthranilates (10), or plastic precursors, such as p-hydroxystyrene (11, 12), yet the TAL reaction may be the limiting step (10, 13).Due to the significant interest in production of phenolic compounds in biotechnological processes, we aimed at increasing the range of characterized phenylalanine ammonia-lyases (PAL) and tyrosine ammonia-lyases (TAL), and in particular at identifying the optimal TAL for use in microbial cell factories...

show abstract

“…The PCR product of PAL cDNA was purified using the QIAquick Gel extraction Kit (Qiagen, Germany), cloned into the pTZ 57R/T Vector, using a Fermentase PCR cloning Kit (Fermentas Inc., Vilnius, Lithuania) and transformed into competent GM2163 E. coli using chilled calcium chloride as recommended previously (Hsieh et al 2010). The Roche High Pure Plasmid Isolation kit was then used to purify the plasmid according to the manufacturer's instructions (Roche Applied Science, Germany).…”

Section: Pal Sequencing and Analysismentioning

confidence: 99%

cDNA cloning, Phylogenic Analysis and Gene Expression Pattern of Phenylalanine ammonia-lyase in Sugarcane (Saccharum officinarum L.)

Hashemitabar

Kolahi

Tabandeh

et al. 2014

Braz. arch. biol. technol.

View full text Add to dashboard Cite

show abstract

Cloning and expression of a phenylalanine ammonia-lyase gene (BoPAL2) from Bambusa oldhamii in Escherichia coli and Pichia pastoris

Cited by 36 publications

References 33 publications

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

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

Highly Active and Specific Tyrosine Ammonia-Lyases from Diverse Origins Enable Enhanced Production of Aromatic Compounds in Bacteria and Saccharomyces cerevisiae

cDNA cloning, Phylogenic Analysis and Gene Expression Pattern of Phenylalanine ammonia-lyase in Sugarcane (Saccharum officinarum L.)

Contact Info

Product

Resources

About