Abstract

Phenylalanine ammonia-lyase [PAL, EC 4.3.1.24 (formerly EC 4.3.1.5)], functions in the plant phenylpropanoid biosynthetic pathway to deaminate the amino acid L-phenylalanine forming trans-cinnamic acid and ammonia. The human inherited metabolic disorder phenylketonuria (PKU) is characterized by an inability of individuals to metabolize phenylalanine. Toward the development of a plant-PAL based therapeutic for the treatment of this disorder, a comparative analysis of PAL activities within various members of the Poaceae was undertaken. This led to the identification of a Zea mays cultivar, Japanese Striped corn with very high levels of PAL specific activity in seedling tissues. The root tissues of this corn variety contain greater levels of PAL gene transcripts and PAL activities, compared to those of the shoot tissues, and are intensely colored due to the accumulation of anthocyanin pigments. PAL activities in the root tissues of young seedlings of another corn variety that lacked root anthocyanins (Indian Blue corn) were generally 30-50% lower than those of Japanese Striped corn seedlings at equivalent growth stages. In general, various stress or hormonal treatments led to minimal changes in PAL specific activity of maize tissues, as compared to controls. The PAL enzymes of Japanese Striped corn root tissues are robust; roots retained 90% of their PAL activity after freeze-drying and >50% activity after freeze-drying and a subsequent 15-week storage at 4 degrees C. This work serves as a prelude to the formulation of a dietary supplement for treatment of PKU based on preserved edible cereal root tissues with high levels of intrinsic PAL activity.

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