Howard Michael B, Ekborg Nathan A, Weiner Ronald M, Hutcheson Steven W
Department of Cell Biology and Molecular Genetics, University of Maryland, Microbiology Bldg, College Park, MD 20742, USA.
J Ind Microbiol Biotechnol. 2003 Nov;30(11):627-35. doi: 10.1007/s10295-003-0096-3. Epub 2003 Nov 11.
Multiple industrial and medical uses of chitin and its derivatives have been developed in recent years. The demand for enzymes with new or desirable properties continues to grow as additional uses of chitin, chitooligosaccharides, and chitosan become apparent. Microorganisms, the primary degraders of chitin in the environment, are a rich source of valuable chitin-modifying enzymes. This review summarizes many methods that can be used to isolate and characterize chitin-modifying enzymes including chitin depolymerases, chitodextrinases, chitin deacetylases, N-acetylglucosaminidases, chitin-binding proteins, and chitosanases. Chitin analogs, zymography, detection of reducing sugars, genomic library screening, chitooligosaccharide electrophoresis, degenerate PCR primer design, thin layer chromatography, and chitin-binding assays are discussed.
近年来，几丁质及其衍生物已在多个工业和医学领域得到应用。随着几丁质、壳寡糖和壳聚糖的更多用途逐渐显现，对具有新的或理想特性的酶的需求持续增长。微生物是环境中几丁质的主要降解者，是有价值的几丁质修饰酶的丰富来源。本综述总结了许多可用于分离和表征几丁质修饰酶的方法，包括几丁质解聚酶、壳糊精酶、几丁质脱乙酰酶、N - 乙酰葡糖胺酶、几丁质结合蛋白和壳聚糖酶。文中还讨论了几丁质类似物、酶谱分析、还原糖检测、基因组文库筛选、壳寡糖电泳、简并PCR引物设计、薄层色谱和几丁质结合测定等方法。
