β-elimination of hyaluronate by red king crab hyaluronidase

• Published in: Scientific Reports
• Main Authors: Sliadovskii, Dmitrii, Ponomareva, Tatyana, Molchanov, Maxim, Pozdnyakova-Filatova, Irina, Timchenko, Maria, Marchenkov, Victor, Gusev, Oleg, Sogorin, Evgeny
• Format: Article in Journal/Newspaper
• Language: English
• Published: Springer Science and Business Media LLC 2021
• Subjects: Multidisciplinary; Red king crab
• Online Access: http://dx.doi.org/10.1038/s41598-021-01890-3; https://www.nature.com/articles/s41598-021-01890-3.pdf; https://www.nature.com/articles/s41598-021-01890-3

Abstract Crustacean hyaluronidases are poorly understood both in terms of their enzymatic properties and in terms of their structural features. In this work, we show that the hepatopancreas homogenate of the red king crab has a hyaluronidase activity that is an order of magnitude higher than its commercial counterpart. Zymography revealed that the molecular weight of a protein with hyalorunidase activity is 40–50 kDa. Analysis of the hepatopancreas transcriptome and results of cloning and sequencing of cDNA revealed a hyaluronidase sequence with an expected molecular weight of 42.5 kDa. Further analysis showed that hyaluronat enzymatic cleavage follows the $$\beta $$ β -elimination mechanism, which is well known for bacterial hyaluronidases. The results of ion-exchange chromatography showed that the final product of hyaluronate degradation is unsaturated tetrasaccharide. Thus, we identified a new hyaluronidase of higher eukaryotes, which is not integrated into the modern classification of hyaluronidases.