Federal Research Center "Pushchino Scientific Center for Biological Research of the RAS", Pushchino, Russia, 142290.
Institute of Theoretical and Experimental Biophysics of the RAS, Pushchino, Russia, 142290.
Sci Rep. 2021 Nov 19;11(1):22600. doi: 10.1038/s41598-021-01890-3.
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 [Formula: see text]-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.
甲壳动物透明质酸酶在酶学性质和结构特征方面都知之甚少。在这项工作中，我们表明，红王蟹的肝胰腺匀浆具有比其商业对应物高一个数量级的透明质酸酶活性。酶谱分析显示，具有透明质酸酶活性的蛋白质的分子量为 40-50 kDa。肝胰腺转录组分析和 cDNA 克隆及测序的结果揭示了一种透明质酸酶序列，其预期分子量为 42.5 kDa。进一步的分析表明，透明质酸酶的酶切遵循[公式：见文本]-消除机制，这种机制为细菌透明质酸酶所熟知。离子交换色谱的结果表明，透明质酸盐降解的最终产物是不饱和四糖。因此，我们鉴定了一种新型的真核生物透明质酸酶，它不属于现代透明质酸酶分类。
