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Author:
Date:
2013-09-05
[Abstract] Endo-β–mannanases in plant require post-translational modification, such as N-glycosylation and disulfide-linked dimerization, for their catalytic activity. Determination of the plant endo-β–mannanase activity needs to modify the assay conditions for optimizing their enzymatic reaction. Here, we describe a modified method for plant endo-β–mannanase assay. A high-salt buffer without thiol reductants is required for effective extraction of the enzyme. The enzyme is able to digest water-insoluble AZCL galactomannan to release water soluble dyed fragments, which is detected through measurement of absorbance at 590 nm wavelength. Increase in absorbance at 590 nm is correlated directly with enzyme activity.
[摘要] 对于其催化活性，植物中的内切-β-甘露聚糖酶需要翻译后修饰，例如N-糖基化和二硫键连接的二聚化。 植物内切-β-甘露聚糖酶活性的测定需要改变测定条件以优化它们的酶反应。 在这里，我们描述了植物内切-β-甘露聚糖酶测定的修改方法。 为了有效地提取酶，需要没有硫醇还原剂的高盐缓冲液。 该酶能够消化水不溶性AZCL半乳甘露聚糖以释放水溶性染色片段，其通过测量在590nm波长处的吸光度来检测。 在590nm处的吸光度的增加与酶活性直接相关
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