Advances and future directions in betalain metabolic engineering

Betalains are the yellow and violet pigments that substitute anthocyanins in plants belonging to the order Caryophyllales. These pigments have attracted much attention because of their bioactivities, which range from an antioxidant capacity to the chemoprevention of cancer. However, the biosynthetic pathway of betalains remains under discussion; the main steps have been characterized in recent years, but multiple side reactions are possible. The key enzymes involved have only recently been described, providing clues about the regulation of betalain biosynthesis. In this review, we provide a comprehensive view of the biosynthetic scheme of betalains and discuss the different reactions that have been demonstrated experimentally or proposed in the literature. Copyright © 2013 Elsevier Ltd. All rights reserved.

Anthocyanins are red and violet pigments that color flowers, fruits and epidermal tissues in virtually all flowering plants. A single order, Caryophyllales, contains families in which an unrelated family of pigments, the betalains, color tissues normally pigmented by anthocyanins. Here we show that CYP76AD1 encoding a novel cytochrome P450 is required to produce the red betacyanin pigments in beets. Gene silencing of CYP76AD1 results in loss of red pigment and production of only yellow betaxanthin pigment. Yellow betalain mutants are complemented by transgenic expression of CYP76AD1, and an insertion in CYP76AD1 maps to the R locus that is responsible for yellow versus red pigmentation. Finally, expression of CYP76AD1 in yeast verifies its position in the betalain biosynthetic pathway. Thus, this cytochrome P450 performs the biosynthetic step that provides the cyclo-DOPA moiety of all red betacyanins. This discovery will contribute to our ability to engineer this simple, nutritionally valuable pathway into heterologous species.