Synthesis and Investigation of the Bio-polymerization of Cutin Monomers and Derivatives

Research output: ResearchPh.D. thesis – Annual report year: 2018

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Cutin and suberin are two plant polyesters whose formation is tightly linked to plants ability to thrive in areal medium due to their ability to restrict gas and water loss and movement. Additionally, these two polymers present properties which make them suitable for the polymer industry. Hence, understanding the bio-formation of these polymers is important for both, fundamental biology as well as for industrial applications. In this context, the identification of cutin synthase 1 (CUS1) in tomato fruit, made possible through a collaborative effort between the group of Professor Jocelyn K. C. Rose at Cornell University and the group of Professor Mads H. Clausen at the Technical University of Denmark, represents an important milestone towards this goal. This enzyme, which is part of the GDSL esterase/lipase superfamily, was proven to catalyze the in vitro oligomerization of 2-mono(10,16-dihydroxyhexadecanoyl)glycerol (2-MHG), the main precursor of tomato cutin. However, up to date, very little is known about CUS enzymes, their selectivity, their tertiary structure and their mechanism of action.

This thesis presents the synthesis of another cutin monomer, a deuterated derivative of this monomer as well as five other 2-MHG derivatives. All these compounds have been synthesized to be used in CUS1-mediated polymerization to gain more information on CUS1 selectivity and mechanism of action. Additionally, the use of one of these compound as a CUS1 ligand in co-crystallization experiments has been initiated. Obtaining CUS1-ligand crystals would be an essential tool towards the discovery of CUS1 tertiary structure through X-ray diffraction (XRD) analysis. The second part of this dissertation discloses a biological approach towards the understanding of CUS1 mechanism of action. In particular, the interaction between CUS1 and its substrates through the formation of hydrophobic tunnels was investigated through the formation of CUS1 mutants through site-directed mutagenesis. Additionally, the investigation of several Arabidopsis GDSL mutants as potential suberin synthase was initiated through the production and characterization of knockdown mutants via RNA silencing.
Original languageEnglish
Place of PublicationLyngby, Denmark
PublisherTechnical University of Denmark (DTU)
Number of pages132
StatePublished - 2018

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