Enzymatic cross-linking of gelatine with laccase and tyrosinase
Enzymatic cross-linking of gelatine with laccase and tyrosinase
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DOI：
10.3109/10242422.2012.646036
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发表时间：
2012-02-01
影响因子：
1.8
通讯作者：
Guebitz, Georg M.
中科院分区：
文献类型：
--
作者：
Jus, Suzana;Stachel, Ines;Guebitz, Georg M.
传统的蛋白质交联涉及有毒化学品的使用。在此，明胶和明胶水解产物与来自钝葡萄座腔菌（Botryosphaeria obtusa）（BoT 1和BoT 2）、双孢蘑菇（Agaricus bisporus）（AbT）和来自刺孢疣微菌（VsT）的酪氨酸酶以及与来自毛栓菌（Trametes hirsuta）（ThL）和T. versicolor（TvL）。酪氨酸残基的酶促氧化通过UV/维斯和荧光光谱法指示，并通过氧消耗测量进一步证实。使用模型底物（Tyr-Ala）的二聚化，证明了通过使用RP-HPLC和LC-MS。酶交联显着增加了可溶性材料的分子量的沉淀点，如通过SDS-PAGE和尺寸排阻色谱法证明。在酚类分子如儿茶素的存在下，交联的效果进一步增强。
Conventional cross-linking of proteins involves the use of toxic chemicals. Here, cross-linking of gelatine and gelatine hydrolysates with tyrosinases from Botryosphaeria obtusa (BoT1 and BoT2), Agaricus bisporus (AbT) and from Verrucomicrobium spinosum (VsT) and with laccases from Trametes hirsuta (ThL) and T. versicolor (TvL) was demonstrated. Enzymatic oxidation of tyrosine residues was indicated by UV/VIS and fluorescence spectroscopy and further confirmed by oxygen consumption measurements. Using a model substrate (Tyr-Ala) dimerization was demonstrated by using RP-HPLC and LC-MS. Enzymatic cross-linking significantly increased the molecular weight of the soluble material up to the point of precipitation as demonstrated by both SDS-PAGE and size exclusion chromatography. The effect of cross-linking was further enhanced in the presence of phenolic molecules such as catechin.