Ligninolytic enzymes from white-rot fungi white-rot fungi are widely used in biotechnological processes. However, the application of these enzymes as free enzymes is limited due to their instability and lack of reusability. Enzyme stabilization is therefore a major challenge in biocatalytic process research, and immobilization methods are desirable. Using cross-linked enzyme aggregates (CLEAs) such as magnetic CLEAs, porous-CLEAs and combi-CLEAs is a promising technique for overcoming these issues. Cross-linking methods can stabilize and immobilize enzymes by interconnecting enzyme molecules via multiple bonds using cross-linking agents such as glutaraldehyde. The high catalyst density and microporous assembly of CLEAs guarantee high catalyst activity, which, together with their long shelf life, operational stability, and reusability, provide a cost-efficient alternative to matrix-assisted immobilization approaches. Here, we review current progress in ligninolytic enzyme immobilization and provide a comprehensive review of CLEAs. Moreover, we summarize the use of these CLEAs for biocatalysis processes, bioremediation such as dye decolourization, wastewater treatment or pharmaceutically active compound elimination.

白腐真菌中的木质素分解酶白腐真菌被广泛应用于生物技术过程中。然而，由于它们的不稳定性和缺乏可重复使用性，限制了这些酶作为游离酶的应用。因此，酶的稳定化是生物催化过程研究中的主要挑战，并且固定化方法是理想的。使用交联的酶聚集体（CLEA），例如磁性CLEA，多孔CLEA和combi-CLEA，是解决这些问题的有前途的技术。交联方法可以通过使用戊二醛等交联剂通过多个键使酶分子相互连接来稳定和固定化酶。CLEA的高催化剂密度和微孔组装确保了高催化剂活性，以及​​长寿命，操作稳定性和可重复使用性，为基质辅助固定方法提供了一种经济高效的替代方法。在这里，我们综述了木质素分解酶固定化的最新进展，并对CLEAs进行了全面的综述。此外，我们总结了这些CLEA在生物催化过程，生物修复（如染料脱色，废水处理或药物活性化合物消除）中的用途。