A Novel Hydroxylation Step in the Taxane Biosynthetic Pathway: A New Approach to Paclitaxel Production by Synthetic Biology

Frontiers in Bioengineering and Biotechnology
Raul Sanchez-MuñozElisabeth Moyano

Abstract

Engineered plant cell lines have the potential to achieve enhanced metabolite production rates, providing a high-yielding source of compounds of interest. Improving the production of taxanes, pharmacologically valuable secondary metabolites of Taxus spp., is hindered by an incomplete knowledge of the taxane biosynthetic pathway. Of the five unknown steps, three are thought to involve cytochrome P450-like hydroxylases. In the current work, after an in-depth in silico characterization of four candidate enzymes proposed in a previous cDNA-AFLP assay, TB506 was selected as a candidate for the hydroxylation of the taxane side chain. A docking assay indicated TB506 is active after the attachment of the side chain based on its affinity to the ligand 3'N-dehydroxydebenzoyltaxol. Finally, the involvement of TB506 in the last hydroxylation step of the paclitaxel biosynthetic pathway was confirmed by functional assays. The identification of this hydroxylase will contribute to the development of alternative sustainable paclitaxel production systems using synthetic biology techniques.

References

Dec 1, 1994·Journal of Computer-aided Molecular Design·Y P Pang, A P Kozikowski
Jul 13, 2000·Journal of Molecular Biology·O EmanuelssonG von Heijne
Feb 15, 2001·Proceedings of the National Academy of Sciences of the United States of America·A SchoendorfR B Croteau
Nov 15, 2001·Proceedings of the National Academy of Sciences of the United States of America·S JenneweinR B Croteau
Jun 8, 2002·Biotechnology Progress·Rosa M CusidóM Teresa Piñol
Sep 17, 2002·Proceedings of the National Academy of Sciences of the United States of America·Kevin WalkerRodney Croteau
Dec 21, 2002·Journal of Experimental Botany·Elisabet MoyanoKirsi-Marja Oksman-Caldentey
May 6, 2003·Archives of Biochemistry and Biophysics·Stefan JenneweinRodney Croteau
Sep 25, 2003·Annual Review of Plant Biology·Mary A Schuler, Daniele Werck-Reichhart
Oct 11, 2003·Biotechnology Advances·S R Rao, G A Ravishankar
Jun 5, 2004·Archives of Biochemistry and Biophysics·MyDoanh Chau, Rodney Croteau
Oct 21, 2004·The Journal of Biological Chemistry·Kevin D WalkerRodney Croteau
Sep 3, 2005·Biochemical and Biophysical Research Communications·Robert M Long, Rodney Croteau
Oct 14, 2005·Journal of Computational Chemistry·James C PhillipsKlaus Schulten
May 23, 2007·Nucleic Acids Research·Markus Wiederstein, Manfred J Sippl
Aug 21, 2007·Current Pharmaceutical Biotechnology·Kham Vongpaseuth, Susan C Roberts
Apr 29, 2009·Journal of Computational Chemistry·Garrett M MorrisArthur J Olson
May 23, 2009·The FEBS Journal·Matteo MagnaniMaurizio Botta
Feb 1, 2006·Phytochemistry Reviews : Proceedings of the Phytochemical Society of Europe·Rodney CroteauMark R Wildung
Jun 1, 2006·Phytochemistry Reviews : Proceedings of the Phytochemical Society of Europe·Rüdiger Kaspera, Rodney Croteau
Aug 24, 2010·Biotechnology Progress·Oscar ExpositoRosa M Cusido
Jan 1, 1990·Methods in Molecular Biology·D R Duncan, J M Widholm
Oct 13, 2011·Molecular Systems Biology·Fabian SieversDesmond G Higgins
Feb 4, 2012·The Arabidopsis Book·Søren BakDanièle Werck-Reichhart
Mar 1, 2012·Bioinformatics·Konstantin OkonechnikovUNKNOWN UGENE team
Nov 28, 2012·Biochemical Engineering Journal·Michael C NaillSusan C Roberts
May 24, 2014·Plant Biotechnology Journal·Miriam OnrubiaAlain Goossens
Jun 10, 2014·Plant Biotechnology Journal·Ana-Belén Sabater-JaraRosa M Cusidó
Jan 1, 2013·Metabolites·Rainer BreitlingEriko Takano
Apr 8, 2015·Nucleic Acids Research·Weizhong LiRodrigo Lopez
Apr 23, 2015·Plant Biotechnology Journal·Karla Ramírez-EstradaJavier Palazon
May 8, 2015·Nature Protocols·Lawrence A KelleyMichael J E Sternberg
Aug 27, 2016·Briefings in Bioinformatics·Dukka B Kc

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Datasets Mentioned

BETA
KP178208

Methods Mentioned

BETA
transfection
Assay
PCR

Software Mentioned

ACD
Clustal Omega
Unipro UGENE
PROSAweb
TargetP
TMHMM
AutoDockTools
ChemSketch
NAMD
Phyre2

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