Purification and Characterization of the Enzyme Fucoidanase from Cobetia amphilecti Utilizing Fucoidan from Undaria pinnatifida

Abstract

Fucoidanase is an unstable enzyme with high specificity that requires a large about of time to screen it from microorganisms. In this study, enzymatic hydrolysis was used to produce low-molecular-weight fucoidan from microorganisms via the degradation of high-molecular-weight fucoidan without damage to the sulfate esterification structure of oligosaccharide. The microbial strain HN-25 was isolated from sea mud and was made to undergo mutagenicity under ultraviolet light. Fucoidanase was extracted via ultrasonication and its enzymatic activity was improved via optimization of the ultrasonic conditions. The enzymatic properties and degradation efficiency of fucoidanase were characterized. The microbial strain HN-25 is a Gram-negative aerobic and rod-shaped-cell bacterium, and therefore was identified as Cobetia amphilecti via 16s rDNA. The results proved that fucoidanase is a hydrolytic enzyme with a molecular weight of 35 kDa and with high activity and stability at 30 °C and pH 8.0. The activity of fucoidanase was significantly enhanced by sodium and calcium ions and inhibited by a copper ion and ethylenediaminetetraacetate (EDTA). There was a significant decrease in the molecular weight of fucoidan after enzymatic hydrolysis. The low-molecular-weight fuicodan was divided into four fractions, mainly concentrated at F3 (20~10 kDa) and F4 (≤6 kDa). These consequences suggest that fucoidanase obtained from Cobetia amphilecti is stable and efficient and could be a good tool in the production of bioactive compounds.

Leucine rich repeat-malectin receptor kinases IGP1/CORK1, IGP3 and IGP4 are required for arabidopsis immune responses triggered by β-1,4-D-Xylo-oligosaccharides from plant cell walls

“…The observation that Arabidopsis thaliana recognizes β-1,6-D-Glc oligosaccharides (purified here for the first time from fungal pustulan), as well as fucoidan and low molecular weight-alginates ( Figs. S1 and S2 ) is in line with the previous description of the effect of these compounds eliciting defensive and abiotic stress responses in different plant species ( Aitouguinane et al, 2020 , Aitouguinane et al, 2023 , Liu et al, 2023 ), and indicates that their mechanisms of perception (e.g. PRRs) are conserved in Arabidopsis thaliana .…”

“…S2 ), D-Glucuronyl in XUXX ( Fig. 1 ) and GalA in OGs ( Galletti et al, 2008 , Voxeur et al, 2019 , Liu et al, 2023 ). Also, Arabidopsis thaliana perceives glycan structures containing L-monosaccharides (L-Arabinose in arabinoxylans, L-Fucose in fucoidans and L-guluronic acid (G), in alginates: Fig.…”

“…These data are also in line with the recent demonstration of the activity of mixtures of oligosaccharides from xylan hydrolysis on crop protection ( Dewangan et al, 2023 , Pring et al, 2023 ). Similarly, several studies have reported the use of alginate in agriculture to stimulate plant growth and development ( Liu et al, 2023 ) and to induce plant resistance mechanisms, but the molecular bases of these responses have not been determined ( Klarzynski et al, 2000 , Aitouguinane et al, 2020 , Aitouguinane et al, 2023 , Wang et al, 2023 ). Together these results confirm the potential of glycans to activate PTI and to develop biological products that boost crop natural defence against pathogens, contributing to replacing chemical pesticides and implementing more sustainable agriculture practices.…”