Department of Chemistry, Virginia Tech, Blacksburg, Virginia24061, United States.
Macromolecules Innovation Institute, Virginia Tech, Blacksburg, Virginia24061, United States.
Biomacromolecules. 2023 Feb 13;24(2):566-575. doi: 10.1021/acs.biomac.2c00538. Epub 2023 Jan 30.
Changes in mass and viscoelasticity of chitin layers in fungal cell walls during chitinase attack are vital for understanding bacterial invasion of and human defense against fungi. In this work, regenerated chitin (RChitin) thin films mimicked the fungal chitin layers and facilitated studies of degradation by family 18 chitinases from () and family 19 chitinases from () that possessed chitin-binding domains (CBDs) that were absent in the family 18 chitinases. Degradation was monitored via a quartz crystal microbalance with dissipation monitoring (QCM-D) in real time at various pH and temperatures. Compared to substrates of colloidal chitin or dissolved chitin derivatives and analogues, the degradation of RChitin films was deeply affected by chitinase adsorption. While the family 18 chitinases had greater solution activity on chitin oligosaccharides, the family 19 chitinases exhibited greater surface activity on RChitin films, illustrating the importance of CBDs for insoluble substrates.
在几丁质酶攻击过程中真菌细胞壁中几丁质层的质量和粘弹性变化对于理解细菌入侵和人体防御真菌至关重要。在这项工作中,再生几丁质(RChitin)薄膜模拟了真菌几丁质层,并促进了对来自()的家族 18 几丁质酶和来自()的家族 19 几丁质酶的降解研究,这些酶具有家族 18 几丁质酶中不存在的几丁质结合域(CBD)。通过石英晶体微天平实时监测在不同 pH 和温度下的降解情况。与胶体几丁质或溶解的几丁质衍生物和类似物的底物相比,RChitin 薄膜的降解受到几丁质酶吸附的深度影响。虽然家族 18 几丁质酶在几丁寡糖上具有更大的溶液活性,但家族 19 几丁质酶在 RChitin 薄膜上表现出更大的表面活性,这表明 CBD 对于不溶性底物的重要性。