Bisepoxide Cross‐Linked Enzyme Aggregates—New Immobilized Biocatalysts for Selective Biotransformations
Bisepoxide Cross‐Linked Enzyme Aggregates—New Immobilized Biocatalysts for Selective Biotransformations
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DOI：
10.1002/cctc.201300806
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发表时间：
2014-05
期刊：
影响因子：
4.5
通讯作者：
Diána Weiser;A. Varga;K. Kovács;F. Nagy;A. Szilágyi;B. Vértessy;Csaba Paizs;L. Poppe
中科院分区：
文献类型：
--
作者：
Diána Weiser;A. Varga;K. Kovács;F. Nagy;A. Szilágyi;B. Vértessy;Csaba Paizs;L. Poppe
甘油二缩水甘油醚(GDE)是一种方便、廉价的双环氧化物交联剂，从两类酶制备的交联酶聚集体(CLEA)中得到了证明。荧光假单胞菌脂肪酶(AK)、洋葱伯克霍尔德氏菌脂肪酶(PS)、南极假丝酵母菌脂肪酶B(CalB)以及巴西石油公司的苯丙氨酸解氨酶(PAL)的GDE酶比它们的戊二醛(GA)交联酶具有更好的性质。超声研究表明，脂肪酶PS和PAL的GDE CLEA在机械上比GA CLEA更稳定。在RAc-1-苯乙醇的动力学拆分中，22℃下制备的GDE-脂肪酶CLEA对AK、CalB和PS的催化活性分别为69.6、134.8和127.4 U g−1，高于相应的GA-脂肪酶CLEA(22℃下制备的Gde-脂肪酶对AK、CalB和PS的催化活性分别为24.4、131.0和119.2 U g−1)。从PAL和牛血清白蛋白(BSA)中提取的GDE共Cleas在6M氨水中与(E)-3-(2-硫酚-2-基)丙烯酸的立体选择性氨加成反应至少可循环3次，而传统的GA-PAL Cleas不能从该介质中回收。
Glycerol diglycidyl ether (GDE) is a convenient and inexpensive bisepoxide cross‐linker as demonstrated by the preparation of cross‐linked enzyme aggregates (CLEAs) from two enzyme classes. The GDE CLEAs of lipase from Pseudomonas fluorescens (AK), lipase from Burkholderia cepacia (PS), and lipase B from Candida antarctica (CaL B) as well as of phenylalanine ammonia‐lyase (PAL) from Petroselinum crispum demonstrated improved properties as compared with their glutaraldehyde (GA) cross‐linked counterparts. Ultrasonication studies indicated that the GDE CLEAs of lipase PS and PAL were mechanically more stable than the GA CLEAs. In the kinetic resolution of rac‐1‐phenylethanol, the catalytic activity of the GDE–lipase CLEAs (U=69.6, 134.8, and 127.4 U g−1 for AK, CaL B, and PS prepared at 22 °C, respectively) surpassed that of the corresponding GA–lipase CLEAs (U=24.4, 131.0, and 119.2 U g−1 for AK, CaL B, and PS prepared at 22 °C, respectively). The GDE co‐CLEAs from PAL and bovine serum albumin (BSA) could be recycled at least three times if used for the stereoselective ammonia addition in 6 M ammonia to (E)‐3‐(thiophen‐2‐yl)acrylic acid, whereas the recycling of the conventional GA–PAL CLEAs from this medium failed.