Biochemical Characterization of a Prokaryotic Phenylalanine Ammonia Lyase

Xiang, Longkuan; Moore, Bradley S.

doi:10.1128/jb.187.12.4286-4289.2005

Cited by 109 publications

(63 citation statements)

References 26 publications

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“…The high pH optimum of the reactions may be a reflection of the reaction mechanism and is generally identical for all enzymes and similar to what has been observed previously for RsTAL, SeSam8, and other homologous enzymes (18,20,21,23,27,(78)(79)(80)(81). Alkaline pH may also be used for biocatalysis, where tyrosine produced by fermentation at physiological pH can be converted to pHCA in high titers by subsequent reaction at alkaline pH in the presence of TAL-expressing E. coli cells or cell paste (82).…”

Section: Resultsmentioning

confidence: 73%

“…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.…”

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confidence: 99%

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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

172

183

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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...

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Section: Resultsmentioning

confidence: 73%

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

172

183

View full text Add to dashboard Cite

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“…1) [16,25]. Although plant PALs, such as from Petroselinum crispum, can convert only phenylalanine into cinnamic acid, PALs from Rhodosporidium toruloides, which have been thoroughly studied, can convert tyrosine to p-hydroxy-cinnamic acid, as well as phenylalanine into cinnamic acid [23].…”

Section: Introductionmentioning

confidence: 99%

“…Although plant PALs, such as from Petroselinum crispum, can convert only phenylalanine into cinnamic acid, PALs from Rhodosporidium toruloides, which have been thoroughly studied, can convert tyrosine to p-hydroxy-cinnamic acid, as well as phenylalanine into cinnamic acid [23]. Alternatively, the marine actinomycete Streptomyces maritimus can convert the amino acid L-phenylalanine into cinnamic acid by a novel bacterial phenylalanine ammonia lyase (PAL) in the enterocin type II polyketide synthesis process [16,25,26]. PAL from S. maritimus, having high substrate specificity for L-phenylalanine, allows for efficient cinnamic acid production [25].…”

Section: Introductionmentioning

confidence: 99%

Cinnamic acid production using Streptomyces lividans expressing phenylalanine ammonia lyase

Noda

Miyazaki

Miyoshi

et al. 2011

J Ind Microbiol Biotechnol

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Cinnamic acid production was demonstrated using Streptomyces as a host. A gene encoding phenylalanine ammonia lyase (PAL) from Streptomyces maritimus was introduced into Streptomyces lividans, and its expression was confirmed by Western blot analysis. After 4 days cultivation using glucose as carbon source, the maximal level of cinnamic acid reached 210 mg/L. When glycerol (30 g/L) was used as carbon source, the maximal level of produced cinnamic acid reached 450 mg/L. In addition, using raw starch, xylose or xylan as carbon source, the maximal level of cinnamic acid reached 460, 300, and 130 mg/L, respectively. We demonstrated that S. lividans has great potential to produce cinnamic acid as well as other aromatic compounds.

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“…More recently, PALs from plants [157] and prokaryotic origin [145,158,159] have become more readily available for bioconversion reactions through their overexpression in efficient enzyme production systems. Diversa recently isolated a set of 18 new PALs by a sequence-based approach [160], 16 of which were from bacterial species whose genome sequence was already publicly available and erroneously had been annotated as genes encoding a histidine ammonia lyase.…”

Section: Phenylalanine Ammonia Lyase-catalyzed Production Of L-phenylmentioning

confidence: 99%

Use of Enzymes in the Synthesis of Amino Acids

Sonke

Kaptein

2009

Amino Acids, Peptides and Proteins in Organic Chemistry

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Biochemical Characterization of a Prokaryotic Phenylalanine Ammonia Lyase

Cited by 109 publications

References 26 publications

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

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

Cinnamic acid production using Streptomyces lividans expressing phenylalanine ammonia lyase

Use of Enzymes in the Synthesis of Amino Acids

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