engleski

A proteomic approach to identify proteins involved in betanin synthesis and its regulation

Betanin is a natural plant pigment that belongs to the water-soluble, nitrogen-containing betalains family of pigments. The commercial importance of betalains stem from its frequent usage as natural food colorants and from its strong anti-oxidant and anti-cancer properties. The most important source of betanin is the red beet (Beta vulgaris subsp. vulgaris) root. The biosynthetic pathway of betanin is known, but all proteins involved in its production and regulation are not yet known. To broaden knowledge of proteins involved, a protemic approach was employed. In our laboratory we dispose of three Agrobacterium tumefaciens transformed sugar beet (Beta vulgaris L.) lines. A green primary tumor line was obtained which produced patches of red cells. From green parts, a green tumor line (Tg) was established, while from the red patches a red tumor line (Tr) was established which overproduced pigment betanin. Third, etiolated, line (Tw) is the Tr grown in complete darknes. In darkness betanin production reversibly ceases. The pigment accumulated in the Tr line was identified as betanin by comparing HPLC spectra of methanolic extracts from red beet and Tr tumor tissue. Total proteins were extracted from tumor tissues and separated on a two-dimensional SDS-PAG electrophoresis. The 2D gels of three cell lines were compared for differences in protein expression and the results are discussed. Further steps will include identification of candidate proteins by mass spectometry.

Proteomics; Mass Spectrometry

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