Purification, biochemical characterisation and bioinformatic analysis of recombinant farnesol dehydrogenase from Theobroma cacao
| Autor: | Maizom Hassan, Chyan Leong Ng, Kok Wai Lam, Mahasakthy-Vijeyasri Satyaveanthan, Saidi-Adha Suhaimi, Alias Awang, Noor-Dina Muhd-Noor |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
0301 basic medicine Physiology Farnesol dehydrogenase Plant Science 01 natural sciences Cofactor 03 medical and health sciences chemistry.chemical_compound Affinity chromatography NAD (+) and NADP (+) Dependent Alcohol Oxidoreductases Genetics Coenzyme binding Phylogeny chemistry.chemical_classification Cacao biology Computational Biology Active site Farnesol 030104 developmental biology Enzyme Biochemistry chemistry Juvenile hormone biology.protein lipids (amino acids peptides and proteins) 010606 plant biology & botany |
| Zdroj: | Plant Physiology and Biochemistry. 161:143-155 |
| ISSN: | 0981-9428 |
| DOI: | 10.1016/j.plaphy.2021.01.050 |
| Popis: | The juvenile hormones (JH) in plants are suggested to act as a form of plant defensive strategy especially against insect herbivory. The oxidation of farnesol to farnesoic acid is a key step in the juvenile hormone biosynthesis pathway. We herein present the purification and characterisation of the recombinant Theobroma cacao farnesol dehydrogenase enzyme that catalyses oxidation of farnesol to farnesal. The recombinant enzyme was purified to apparent homogeneity by affinity chromatography. The purified enzyme was characterised in terms of its deduced amino acid sequences, phylogeny, substrate specificity, kinetic parameters, structural modeling, and docking simulation. The phylogenetic analysis indicated that the T. cacao farnesol dehydrogenase (TcFolDH) showed a close relationship with A. thaliana farnesol dehydrogenase gene. The TcFolDH monomer had a large N-terminal domain which adopted a typical Rossmann-fold, harboring the GxxGxG motif (NADP(H)-binding domain) and a small C-terminal domain. The enzyme was a homotrimer comprised of subunits with molecular masses of 36 kDa. The TcFolDH was highly specific to NADP+ as coenzyme. The substrate specificity studies showed trans, trans-farnesol was the most preferred substrate for the TcFolDH, suggesting that the purified enzyme was a NADP+-dependent farnesol dehydrogenase. The docking of trans, trans-farnesol and NADP+ into the active site of the enzyme showed the important residues, and their interactions involved in the substrate and coenzyme binding of TcFolDH. Considering the extensive involvement of JH in both insects and plants, an in-depth knowledge on the recombinant production of intermediate enzymes of the JH biosynthesis pathway could help provide a potential method for insect control. |
| Databáze: | OpenAIRE |
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