Lipoxygenases have proven to be a potential biocatalyst for various industrial applications. However, low catalytic activity, low thermostability, and narrow range of pH stability largely limit its application. Here, a lipoxygenase (LOX) gene from Enterovibrio norvegicus DSM 15893 (EnLOX) was cloned and expressed in Escherichia coli BL21 (DE3). EnLOX showed the catalytic activity of 40.34 U mg⁻¹ at 50 °C, pH 8.0. Notably, the enzyme showed superior thermostability, and wide pH range stability. EnLOX remained above 50% of its initial activity after heat treatment below 50 °C for 6 h, and its melting point temperature reached 78.7 °C. More than 70% of its activity was maintained after incubation at pH 5.0–9.5 and 4 °C for 10 h. In addition, EnLOX exhibited high substrate specificity towards linoleic acid, and its kinetic parameters of V_max, K_m, and K_cat values were 12.42 mmol min⁻¹ mg⁻¹, 3.49 μmol L⁻¹, and 16.86 s⁻¹, respectively. LC-MS/MS analysis indicated that EnLOX can be classified as 13-LOX, due to its ability to catalyze C₁₈ polyunsaturated fatty acid to form 13-hydroxy fatty acid. Additionally, EnLOX could improve the farinograph characteristics and rheological properties of wheat dough. These results reveal the potential applications of EnLOX in the food industry.

The eukaryotic and archaeal translation factor IF5A requires a post-translational hypusine modification, which is catalyzed by deoxyhypusine synthase (DHS) at a single lysine residue of IF5A with NAD⁺ and spermidine as cofactors, followed by hydroxylation to form hypusine. While human DHS catalyzed reactions have been well characterized, the mechanism of the hypusination of archaeal IF5A by DHS is not clear. Here we report a DHS structure from Pyrococcus horikoshii OT3 (PhoDHS) at 2.2 Å resolution. The structure reveals two states in a single functional unit (tetramer): two NAD⁺-bound monomers with the NAD⁺ and spermidine binding sites observed in multi-conformations (closed and open), and two NAD⁺-free monomers. The dynamic loop region V288–P299, in the vicinity of the active site, adopts different positions in the closed and open conformations and is disordered when NAD⁺ is absent. Combined with NAD⁺ binding analysis, it is clear that PhoDHS can exist in three states: apo, PhoDHS-2 equiv NAD⁺, and PhoDHS-4 equiv NAD⁺, which are affected by the NAD⁺ concentration. Our results demonstrate the dynamic structure of PhoDHS at the NAD⁺ and spermidine binding site, with conformational changes that may be the response to the local NAD⁺ concentration, and thus fine-tune the regulation of the translation process via the hypusine modification of IF5A.