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[与多糖和玻璃载体共价结合的聚半乳糖醛酸酶的作用方式及抑制作用]

[Mode of action and inhibition of polygalacturonase covalently bound to polysaccharide and glass carriers].

作者信息

Bock W, Krause M, Göbel H, Anger H, Schawaller H J, Flemming C, Gabert A

出版信息

Nahrung. 1978;22(2):185-97. doi: 10.1002/food.19780220209.

Abstract

Endo-polygalacturonase (EC 3.2.1.15.) from Aspergillus spec. is much changed as far as its mode of action and the interaction with vegetable inhibitors of pectinase (from green beans and cucumbers) are concerned when it is covalently bound to insoluble carriers (Sepharose, cellulose powder, macroporous glass and nonporous ballotinis). Whereas a 2% degradation of substrate by the soluble enzyme caused a 50% decrease of viscosity of citrus pectic acid, the comparable degradation of substrate was increased to a level of about 10% with the investigated polygalacturonase carrier complexes apparently independent of the properties of the carriers and the kind of binding of the enzyme. In contrast to this the higher degradation of substrate of 15 and 20% respectively which was further stated at a 50% decrease of viscosity is unambiguously connected with the carriers and is in direct correlation with the specific activity of the polygalacturonase carrier complexes. Contrary to the soluble enzyme the covalently bound enzyme produces more lower oligomerous galacturonic acids by an exo-mechanism or by multiple attack already at the beginning of the hydrolysis of pectic acid. During the final stage there is an enrichment of trigalacturonic acid besides mono- and digalacturonic acids independent of the state of solution of the enzyme. It could further be stated that the strong inhibition of the soluble endo-polygalacturonase by selected pectinase inhibitors which was described earlier is reduced by degrees with the enzyme covalently bound to the insoluble carriers.

摘要

来自曲霉属的内切多聚半乳糖醛酸酶(EC 3.2.1.15.)在与不溶性载体(琼脂糖、纤维素粉、大孔玻璃和无孔玻璃珠)共价结合后,其作用方式以及与果胶酶植物抑制剂(来自绿豆和黄瓜)的相互作用发生了很大变化。可溶性酶使底物降解2%会导致柑橘果胶酸粘度降低50%,而对于所研究的多聚半乳糖醛酸酶-载体复合物,底物的类似降解增加到约10%的水平,这显然与载体的性质和酶的结合方式无关。与此相反,分别在粘度降低50%时进一步观察到的底物15%和20%的较高降解率与载体明确相关,并且与多聚半乳糖醛酸酶-载体复合物的比活性直接相关。与可溶性酶相反,共价结合的酶在果胶酸水解开始时就通过外切机制或多次攻击产生更多的低聚半乳糖醛酸。在最后阶段,除了单半乳糖醛酸和二半乳糖醛酸外,三半乳糖醛酸也会富集,这与酶的溶液状态无关。还可以指出,先前描述的选定果胶酶抑制剂对可溶性内切多聚半乳糖醛酸酶的强烈抑制作用会随着酶与不溶性载体共价结合而逐渐降低。

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