NDLI: Next generation sequencing in predicting gene function in podophyllotoxin biosynthesis.

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Next generation sequencing in predicting gene function in podophyllotoxin biosynthesis.

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Next generation sequencing in predicting gene function in podophyllotoxin biosynthesis.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Marques, Joaquim V. Kim, Kye-won Lee, Choonseok May, Gregory D. Crow, John A. Lewis, Norman G. Costa, Michael A. Davin, Laurence B.
Description	Author Affiliation: Marques JV ( Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340, USA.)
Abstract	Podophyllum species are sources of (-)-podophyllotoxin, an aryltetralin lignan used for semi-synthesis of various powerful and extensively employed cancer-treating drugs. Its biosynthetic pathway, however, remains largely unknown, with the last unequivocally demonstrated intermediate being (-)-matairesinol. Herein, massively parallel sequencing of Podophyllum hexandrum and Podophyllum peltatum transcriptomes and subsequent bioinformatics analyses of the corresponding assemblies were carried out. Validation of the assembly process was first achieved through confirmation of assembled sequences with those of various genes previously established as involved in podophyllotoxin biosynthesis as well as other candidate biosynthetic pathway genes. This contribution describes characterization of two of the latter, namely the cytochrome P450s, CYP719A23 from P. hexandrum and CYP719A24 from P. peltatum. Both enzymes were capable of converting (-)-matairesinol into (-)-pluviatolide by catalyzing methylenedioxy bridge formation and did not act on other possible substrates tested. Interestingly, the enzymes described herein were highly similar to methylenedioxy bridge-forming enzymes from alkaloid biosynthesis, whereas candidates more similar to lignan biosynthetic enzymes were catalytically inactive with the substrates employed. This overall strategy has thus enabled facile further identification of enzymes putatively involved in (-)-podophyllotoxin biosynthesis and underscores the deductive power of next generation sequencing and bioinformatics to probe and deduce medicinal plant biosynthetic pathways.
ISSN	00219258
e-ISSN	1083351X
Journal	Journal of Biological Chemistry
Issue Number	1
Volume Number	288
Language	English
Publisher	American Society for Biochemistry and Molecular Biology (United States)
Publisher Date	2013-01-04
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Plants, Medicinal Metabolism Podophyllotoxin Biosynthesis Podophyllum Sequence Analysis, DNA Amino Acid Sequence Catalysis Computational Biology Cytochrome P-450 Enzyme System Databases, Factual Gene Expression Regulation, Plant Lignans Chemistry Microsomes Models, Biological Models, Chemical Molecular Sequence Data Plant Extracts Sequence Homology, Amino Acid Transcriptome Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. Biochemistry Molecular Biology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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