Deutsches Textilforschungszentrum Nord-West gGmbH, 47798 Krefeld, Germany.
Justus Liebig University, Food Chemistry and Food Biotechnology, 35392 Giessen, Germany.
J Dairy Sci. 2021 Feb;104(2):1548-1559. doi: 10.3168/jds.2019-17110. Epub 2020 Dec 11.
Textiles represent promising support materials for enzymes. The goal of the present work was to investigate the immobilization of commercial peroxidase on a polyester needle felt and the repeated use in the gentle degradation of norbixin in whey from dairy cheese as a practical application. High enzyme loads were obtained by a 2-step immobilization procedure. First, the number of functional groups on the textile surface was increased by a modification with amino-functional polyvinylamine. Second, the enzyme was immobilized by using 2 types of crosslinking agents. Due to the iron content of peroxidase, inductively coupled plasma-optical emission spectrometry was used for the quantitative determination of the enzyme load on the textile. The enzyme activity was evaluated using common 2,2'-azino-di-(3-ethylbenzthiazoline-6-sulfonic acid) assay for peroxidases. By the variation of enzyme input and crosslinker concentration, a maximal enzyme load of 80 mg/g of textile was achieved, and a maximum specific activity of 57 U/g of textile. For the visualization of the enzyme on the fiber surface, fluorescence microscopy as well as scanning probe microscopy were used. The immobilized peroxidase showed significant activity, even after 50 reuse cycles. In addition, the potential of the new support and enzyme combination in commercial whey bleaching was demonstrated successfully on a 10-L scale.
纺织品是一种很有前途的酶支持材料。本工作的目的是研究将商业过氧化物酶固定在聚酯针刺毡上，并将其在温和降解乳清中的核黄素中的重复使用作为实际应用。通过两步固定化程序获得了高酶负载量。首先，通过用氨基官能化的聚乙烯亚胺修饰来增加纺织品表面的官能团数量。其次，通过使用 2 种交联剂将酶固定化。由于过氧化物酶的铁含量，使用电感耦合等离子体-光学发射光谱法对纺织品上的酶负载量进行定量测定。通过常用的过氧化物酶的 2,2'-联氮-二-(3-乙基苯并噻唑啉-6-磺酸)测定法评估酶活性。通过改变酶输入和交联剂浓度，可以实现 80mg/g 纺织品的最大酶负载量和 57U/g 纺织品的最大比活度。为了在纤维表面可视化酶，使用了荧光显微镜和扫描探针显微镜。固定化过氧化物酶即使在 50 次重复使用后仍表现出显著的活性。此外，还成功地在 10L 规模上展示了新的支撑材料和酶组合在商业乳清漂白中的潜力。
