β-elimination of hyaluronate by red king crab hyaluronidase
| Autor: | Maria Timchenko, Dmitrii Sliadovskii, Oleg Gusev, Irina Pozdnyakova-Filatova, Evgeny Sogorin, Maxim Molchanov, Tatyana Ponomareva, Victor Marchenkov |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
DNA
Complementary Magnetic Resonance Spectroscopy Science Carbohydrates Hepatopancreas Hyaluronoglucosaminidase Biochemistry Article Substrate Specificity Transcriptome Functional clustering Hyaluronidase Polysaccharides Complementary DNA medicine Tetrasaccharide Protein analysis Animals Zymography Hyaluronic Acid Databases Protein Phylogeny Cloning chemistry.chemical_classification Animal biotechnology Multidisciplinary biology Hydrolysis biology.organism_classification Chromatography Ion Exchange Enzymes Computational biology and bioinformatics Enzyme chemistry Sequence annotation Enzyme mechanisms Biocatalysis Medicine Anomura medicine.drug Biotechnology |
| Zdroj: | Scientific Reports Scientific Reports, Vol 11, Iss 1, Pp 1-11 (2021) |
| ISSN: | 2045-2322 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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