Research of Multicopper Oxidase and Its Degradation of Histamine in LPZN19.

In order to explore the structural changes and products of histamine degradation by multicopper oxidase (MCO) in LPZN19, a 1500 bp gene in LPZN19 was cloned, and the recombinant MCO was expressed in BL21 (DE3). After purification by Ni-NTA affinity chromatography, the obtained MCO has a molecular weight of 58 kDa, and it also has the highest enzyme activity at 50 °C and pH 3.5, with a relative enzyme activity of 100%, and it maintains 57.71% of the relative enzyme activity at 5% salt concentration. The secondary structure of MCO was determined by circular dichroism, in which the proportions of the α-helix, β-sheet, β-turn and random coil were 2.9%, 39.7%, 21.2% and 36.1%, respectively. The 6xj0.1.A with a credibility of 68.21% was selected as the template to predict the tertiary structure of MCO in LPZN19, and the results indicated that the main components of the tertiary structure of MCO were formed by the further coiling and folding of a random coil and β-sheet. Histamine could change the spatial structure of MCO by increasing the content of the α-helix and β-sheet. Finally, the LC-MS/MS identification results suggest that the histamine was degraded into imidazole acetaldehyde, hydrogen peroxide and ammonia.