Optimisation ofRamonda serbicaLEA protein production inEscherichia coliand its secondary structure analysis

• DOI: 10.1101/2025.02.19.639015
• Publication Date: 2025-02-22T03:20:27Z
• Authors: Pantelić Ana; Ilina Tatiana; Milić Dejana; Gradišar Helena; Radosavljević Jelena; Marija Vidović

Desiccation, an extreme form of dehydration, reduces the cellular water content to below 5 % and poses a major challenge for most plants. Ramonda serbica, tertiary relict homoiochlorophyllous resurrection plant, is exceptional model investigating vegetative desiccation tolerance. Late Embryogenesis Abundant (LEA) proteins are strongly involved in this adaptive trait, but their exact molecular function still unclear. In study, we report first successful recombinant production desiccation-induced LEA protein, RsLEAP30, from dicotyledonous plant species using Escherichia coli expression system. By employing immobilised metal affinity size-exclusion chromatography, achieved purify RsLEAP30 purity over 95 %, providing robust scalable method producing other proteins. Structural characterisation by circular dichroism spectroscopy, combined with silico modelling, revealed that predominantly disordered contains alpha-helical regions. We suggest structural duality underpins protective role chloroplasts, likely via interactions thylakoids desiccation-sensitive within photosynthesis-associated This may be crucial rapid recovery photosynthetic components upon rehydration. Our study provides new insights into structure-function relationship plants establishes foundation future investigations. Understanding mechanisms will pave way bioengineering strategies aimed at improving drought tolerance crops.