Enzyme amino acid ammonia lyase

Many amino acids can lose ammonia to give an unsaturated acid. The enzymes that catalyse these reactions are known as amino acid ammonia lyases. The one that concerns us at the end of the shikimic acid pathway is phenylalanine ammonia lyase, which catalyses the elimination of ammonia from phenylalanine to give the common metabolite cinnamic acid. 

In higher plants the polymer, lignin, and various aromatic secondary metabolites, notably many alkaloids (Chapter 6) and flavonoids (Section 5.4) are formed from the aromatic amino acids, L-phenylalanine (5.77) and/or L-tyrosine (5.75). [For some alkaloids as well as some microbial metabolites, tryptophan (5.7 ) is the source of their particular aromatic rings.] There are for these metabolites common pathways leading from phenylalanine, and in some plants tyrosine, to phenylpropanoid (Cg-Cg) intermediates. The first step from phenylalanine involves the enzyme L-phenylalanine ammonia lyase (known, perhaps affectionately, as PAL), an enzyme widely distributed, and well-characterized. Elimination of ammonia occurs to give cinnamic acid 5.23). It involves loss of the (3-/ ro-5)-proton of L-phenylalanine (5.77), and thus occurs in the aw z-sense [L-tyrosine ammonia lyase functions to remove also the (3-/ ro-5)-proton in tyrosine] [12, 13]. 

A PNH3 electrode covered with this enzyme is used [123] for the determination of L-phenylalanine with very little interference fiom otho amino acids, notably L-tyrosine. A similar selectivity is obtained with phenylalanine decarboxylase [103] which, when attached to a pC02 electrode, enables the specific determination of phenylalanine. Other ammonia lyases are used te determine amino acids. Methionine lyase [124] and histidine ammonia lyase [12S] produce ammonia which is detected with a PNH3 electrode. 

Phenylalanine ammonia-lyase (PAL EC 4.3.1.5) is a pivotal enzyme in controlling flow of carbon from aromatic amino acids to secondary aromatic compounds (Figure 1) (28). PAL primarily deaminates phenylalanine to form t-cinnamic acid, however, in many species, it also less efficiently deaminates tyrosine to form -coumaric acid. Because PAL is restricted to plants and is an important enzyme in plant development, Jangaard (29) suggested that PAL inhibitors might make safe and effective herbicides, however, in his screen of several herbicides, he found no compound to have a specific effect on PAL. This was also the case in studies by Hoagland and Duke (30, 31.) in which 16 herbicides were screened. 

This enzyme [EC 4.1.99.1], also known as L-tryptophan indole-lyase, catalyzes the hydrolysis of L-tryptophan to generate indole, pyruvate, and ammonia. The reaction requires pyridoxal phosphate and potassium ions. The enzyme can also catalyze the synthesis of tryptophan from indole and serine as well as catalyze 2,3-elimination and j8-replacement reactions of some indole-substituted tryptophan analogs of L-cysteine, L-serine, and other 3-substituted amino acids. 

Phenylalanine Ammonia-Lyase. The building units of lignin are formed from carbohydrate via the shikimic acid pathway to give aromatic amino acids. Once the aromatic amino acids are formed, a key enzyme for the control of lignin precursor synthesis is phenylalanine ammonia-lyase (PAL) (1). This enzyme catalyzes the production of cinnamic acid from phenylalanine. It is very active in those tissues of the plant that become lignified and it is also a central enzyme for the production of other phenylpropanoid-derived compounds such as flavonoids and coumarins, which can occur in many parts of the plant and in many different organs (35). Radioactive phenylalanine and cinnamic acid are directly incorporated into lignin in vascular tissue (36). 

When soybean leaves and pine needles were exposed to ozone, there was an initial decrease in the levels of soluble sugars followed by a subsequent increase. Ozone exposure also caused a decrease in the activity of the glycolytic pathway and the decrease in the activity was reflected in a lowered rate of nitrate reduction. Amino acids and protein also accumulated in soybean leaves following exposure. Ozone increased the activities of enzymes involved in phenol metabolism (phenylalanine ammonia lyase and polyphenoloxidase). There was also an increase in the levels of total phenols. Leachates from fescue leaves exposed to ozone inhibited nodulation. 

Coumaroyl-CoA is produced from the amino acid phenylalanine by what has been termed the general phenylpropanoid pathway, through three enzymatic conversions catalyzed by phenylalanine ammonia-lyase (PAL), cinnamate 4-hydroxylase (C4H), and 4-coumarate CoA ligase (4CL). Malonyl-CoA is formed from acetyl-CoA by acetyl-CoA carboxylase (ACC) (Figure 3.2). Acetyl-CoA may be produced in mitochondria, plastids, peroxisomes, and the cytosol by a variety of routes. It is the cytosolic acetyl-CoA that is used for flavonoid biosynthesis, and it is produced by the multiple subunit enzyme ATP-citrate lyase that converts citrate, ATP, and Co-A to acetyl-CoA, oxaloacetate, ADP, and inorganic phosphate. ... 

Lyases are an attractive group of enzymes from a commercial perspective, as demonstrated by then-use in many industrial processes.240 They catalyze the cleavage of C-C, C-N, C-O, and other bonds by means other than hydrolysis, often forming double bonds. For example, two well-studied ammonia lyases, aspartate ammonia lyase (aspartase) (E.C. 4.3.1.1) and phenylalanine ammonia lyase (PAL) (E.C. 4.3.1.5), catalyze the trans-elimination of ammonia from the amino acids, l-aspartate and L-phenylalanine, respectively. Most commonly used in the synthetic mode, the reverse reaction has been used to prepare the L-amino acids at the ton scale (Schemes 19.30 and 19.31).240 242 These reactions are conducted at very high substrate concentrations such that the equilibrium is shifted, resulting in very high conversion to the amino acid products. 

Figure 7.16 Phylogenetic relationships in key enzymes of pathways leading to SM, based on amino acid sequences, (a) Ornithine decarboxylase (ODC). (b) Tyrosine decarboxylase (TyrDC). (c) Tryptophan decarboxylase (TDC). (d) Phenylalanine ammonia-lyase (PAL). Numbers at nodes are bootstrap values.

Various commercial routes for the production of L-phenyalanine have been developed because of the utilization of this amino acid in the dipeptide sweetener Aspartame. One route that has been actively pursued is the synthesis of L-phenylalanine from trans-cinnamic acid using the enzyme phenylalanine ammonia lyase (105,106). This enzyme catalyzes the reversible, nonoxidative deamination of L-phenylalanine and can be isolated from various plant and microbial sources (107,108). 

L-Phenylalanine,which is derived via the shikimic acid pathway,is an important precursor for aromatic aroma components. This amino acid can be transformed into phe-nylpyruvate by transamination and by subsequent decarboxylation to 2-phenylacetyl-CoA in an analogous reaction as discussed for leucine and valine. 2-Phenylacetyl-CoA is converted into esters of a variety of alcohols or reduced to 2-phenylethanol and transformed into 2-phenyl-ethyl esters. The end products of phenylalanine catabolism are fumaric acid and acetoacetate which are further metabolized by the TCA-cycle. Phenylalanine ammonia lyase converts the amino acid into cinnamic acid, the key intermediate of phenylpropanoid metabolism. By a series of enzymes (cinnamate-4-hydroxylase, p-coumarate 3-hydroxylase, catechol O-methyltransferase and ferulate 5-hydroxylase) cinnamic acid is transformed into p-couma-ric-, caffeic-, ferulic-, 5-hydroxyferulic- and sinapic acids,which act as precursors for flavor components and are important intermediates in the biosynthesis of fla-vonoides, lignins, etc. Reduction of cinnamic acids to aldehydes and alcohols by cinnamoyl-CoA NADPH-oxido-reductase and cinnamoyl-alcohol-dehydrogenase form important flavor compounds such as cinnamic aldehyde, cin-namyl alcohol and esters. Further reduction of cinnamyl alcohols lead to propenyl- and allylphenols such as... 

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