The coimmobilization of D-amino acid oxidase and catalase enables the quantitative transformation of D-amino acids (D-phenylalanine) into α-keto acids (phenylpyruvic acid)
The coimmobilization of D-amino acid oxidase and catalase enables the quantitative transformation of D-amino acids (D-phenylalanine) into α-keto acids (phenylpyruvic acid)
复制标题
DOI：
10.1016/s0141-0229(98)00028-3
复制
发表时间：
1998-07-01
影响因子：
3.4
通讯作者：
Guisán, JM
中科院分区：
文献类型：
--
作者：
Fernández-Lafuente, R;Rodriguez, V;Guisán, JM
本文研究了可变胡芦巴D-氨基酸氧化酶催化D-苯丙氨酸(D/L外消旋混合物)温和立体定向氧化生成苯丙酮酸。这一反应的进行需要氧气的持续鼓泡，以使可溶性DAAO失活。酶在多孔载体内的固定化避免了酶与疏水的氧/水界面的相互作用；因此，这种失活原因完全避免了过氧化氢(反应的副产物)对酶和所需产品(苯丙酮酸)产生有害影响。使用固定化的DAAO和单独固定化的过氧化氢酶可以减少这些有害影响；然而，当两种酶共同固定化时，过氧化氢的清除要有效得多。共固定化过氧化氢酶防止过氧化氢破坏苯丙酮酸的效果比单独固定化过氧化氢酶高20倍以上。实际上，使用DAAO/过氧化氢酶共固定化的衍生物，以外消旋混合物为底物，D-苯丙氨酸完全氧化后，苯丙酮酸的产率可达98%以上。这也防止了酶的失活。用这种方法进行了40个反应循环，没有明显的酶活性损失。这些结果开辟了一种使用任何酶的有效和简单的方法，其中我们可以预料到与本文(C)1998 Elsevier Science Inc.中描述的问题非常相似的问题。
We have studied the production of phenylpyruvic acid by mild stereospecific oxidation of D-phenylalanine (from D/L racemic mixtures) catalyzed by D-amino acid oxidase (DAAO) from Trigonopsis variabilis. The performance of this reaction requires the continuous bubbling of oxygen that inactivated soluble DAAO. The immobilization of the enzyme inside porous supports avoids the interaction of the enzyme with the hydrophobic oxygen/water interfaces; therefore, this inactivation cause is fully prevented Hydrogen per-oxide la reaction by-product) exerts deleterious effects on both the enzyme and the desired product (phenylpyruvic acid). These deleterious effects could be reduced by using immobilized DAAO and separately immobilized catalase; however, a much more effective elimination of hydrogen peroxide was achieved when both enzymes were coimmobilized. The effectiveness of coimmobilized catalase in preventing phenylpyruvic destruction by hydrogen peroxide is more than 20-fold higher than that of separately immobilized catalase. In fact, by using DAAO/catalase-coimmobilized derivatives, more than 98% phenylpyruvic acid was obtained after complete oxidation of D-phenylalanine by using the racemic mixture as substrate. This also prevented enzyme from inactivation. In this way, 40 reaction cycles were performed without apparent loss of enzyme activity. These results open an effective and simple way of using any oxidase where we can expect very similar problems to those described in this paper (C) 1998 Elsevier Science Inc.