alexa Molecular investigations of Piper nigrum (Black pepper) fruits in search for natural products biosynthetic target genes.
General Science

General Science

Journal of Forensic Biomechanics

Author(s): Cotinguiba F, Manke K, Furlan M, Vogt T

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Phenylpropanoid derivatives, especially amides, are the main compounds found in several Piper species. Compounds like piperine show important biological activities as antitumor, antifungal, insecticidal and trypanocidal. Among the amides isolated from black pepper, the most important compound responsible for the spicy taste is piperine. The biosynthesis of piperine and related compounds is not yet understood partly due to the limited availibility of plant material. In an initial step to investigate the biosynthesis and molecular biology of Piper phenylpropanoids we describe the construction of a cDNA library from total RNA from P. nigrum and the identifi cation of a limited set of genes encoding enzymes both primary and secondary metabolism. For choosing the appropriate tissue for RNA extraction, the concentration of piperine was monitored in various plant tissues (leaves, seeds, stems) using HPLC-UV analysis. Piperine is concentrated only in fruits and virtually absent in other organs, indicating that the enzymatic machinery responsible for the biosynthesis of this compound should be restriced tothe berries only. Therefore, total RNA was extracted from this organ. Preparation of the cDNA (RT-PCR) was followed by construction of a (full length) cDNA library. Based on length classifi cation by agarose gel analysis DNA-sequencing was realized, providing initial information about the trancriptome of Piper nigrum fruits. The sequences were compared and identifi ed through BLAST program (www.ncbi.nlm.nih.gov/BLAST). Thorough investigation of the transcriptome by new “deep sequencing” techniques of Piper fruits will enable us in the future to understand the biosynthetic pathways involved in the formation of piperine and related amides, and will result ultimately in the identifi cation and functional characterisation of the key enzymes. These data may not only be used for biotechnological production of natural products but could facilitate the design and bioorganic synthesis of new molecules for industrial and pharmaceutical use. Financial Support: Fernando Cotinguiba, Maysa Furlan, and Thomas Vogt thank FAPESP, DAAD, and CAPES for the scholarship and fi nancial support

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This article was published in CongressoBrasileirodeGenetica and referenced in Journal of Forensic Biomechanics

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