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Chitosans with defined characteristics have been shown to possess reproducible bioactivities for numerous applications. A promising approach for producing chitosans with defined degrees of polymerization (DP), degrees of acetylation (DA), and patterns of acetylation (PA) involves using chitin-modifying enzymes. One such enzyme, the chitinase Tv-ECH1 belonging to the glycoside hydrolase (GH) family 18, seems to have an important role in the biocontrol properties of the fungus Trichoderma virens, suggesting its potential in generating novel chitosans for plant health applications. In this study, the Tv-ECH1 enzyme was overexpressed in the methylotrophic yeast Pichia pastoris, yielding large amounts (up to 2mgmL-1) of purified recombinant enzyme of high activity, high purity, and high stability, making the system promising for industrial production of Tv-ECH1. The purified Tv-ECH1 chitinase displayed a wide optimal pH range from 4.5 to 6 and an optimal temperature of 37°C. Detailed subsite specificity analyses revealed high preference for acetylated residues at all four subsites analyzed (-2, -1, +1, +2), making Tv-ECH1 a promising candidate for the biotechnological production of specific chitosan oligomers and for the characterization of chitosan polymers via enzymatic fingerprinting.

Global changes in climate are leading to increased occurrence and duration of drought episodes with concurrent reduction in crop yields. Expansion of the irrigated land area does not appear to be a viable solution in many regions to deliver crop productivity. The development of crop drought tolerance traits by either genetic modification or plant breeding represent the principal approaches to meeting this challenge to date. Biostimulants are an emerging category of crop management products which can enhance crop productivity under abiotic stress conditions. The ability of some biostimulant products such as Ascophyllum nodosum extracts (ANE) to enhance the tolerance of crops to drought stress has been observed by growers. The objective of this study was to investigate if different commercial ANE biostimulants provided the same tolerance to tomato plants (cv. Moneymaker) subjected to a defined drought period. A compositional characterisation of the key macromolecules of ANEs was performed. In addition, the role of ANE biostimulants in inducing changes of chlorophyll and osmolytes levels, MDA production, dehydrin isoform pattern and dehydrin gene expression levels was assessed. The three ANE biostimulants evaluated were found to provide different levels of tolerance to drought stressed tomato plants. The level of drought tolerance provided was related to changes in the concentration of osmolytes and expression of tas14 dehydrin gene. Taken together, our results highlight that despite the fact all ANE biostimulants were manufactured from the same raw material, their ability to maintain crop productivity during and after drought stress was not the same.