Singh Pinki Kumari, Pachaiappan Raman
Department of Biotechnology, School of Bioengineering, College of Engineering and Technology, SRM Institute of Science and Technology, Chengalpattu, Kattankulathur, Tamil Nadu, India.
Methods Mol Biol. 2025;2917:163-179. doi: 10.1007/978-1-0716-4478-2_15.
Chitinases are a variety of different enzymes that facilitate the breakdown of chitin, a complex sugar molecule presents in the outer shells of arthropods and the cellular walls of fungi. Chitinase zymography is a powerful analytical technique to visualize and characterize chitinase activity in complex biological samples. In plants, chitinases contribute to defense mechanisms by degrading fungal cell walls and deterring pathogen invasion. They also influence plant growth and development by modulating interactions with soil microbes. This chapter provides different zymographic protocols for chitinase detection, including those using polyacrylamide gel electrophoresis (PAGE) with various chitin derivatives and staining methods. Calcofluor White M2R, is offering unique advantages for visualizing chitinase activity on native PAGE overlayed chitin amended gels. This overview highlights the importance of selecting appropriate substrates and assay conditions to achieve accurate and reliable results in chitinase zymography, thereby advancing the potential of chitinase-based technologies in industrial and environmental fields.
几丁质酶是多种不同的酶，可促进几丁质的分解，几丁质是一种复杂的糖分子，存在于节肢动物的外壳和真菌的细胞壁中。几丁质酶活性电泳图谱是一种强大的分析技术，可用于可视化和表征复杂生物样品中的几丁质酶活性。在植物中，几丁质酶通过降解真菌细胞壁和阻止病原体入侵来促进防御机制。它们还通过调节与土壤微生物的相互作用来影响植物的生长和发育。本章提供了用于检测几丁质酶的不同活性电泳图谱方案，包括那些使用聚丙烯酰胺凝胶电泳（PAGE）以及各种几丁质衍生物和染色方法的方案。荧光增白剂M2R在可视化天然PAGE覆盖的几丁质改良凝胶上的几丁质酶活性方面具有独特优势。本概述强调了选择合适的底物和测定条件以在几丁质酶活性电泳图谱中获得准确可靠结果的重要性，从而提高基于几丁质酶的技术在工业和环境领域的潜力。
