The industrial use of enzymes often requires their immobilization to facilitate downstream processing and enable reuse. However, controlling enzyme orientation during immobilization is challenging and typically restricted to the N- and C-terminal regions. In this work, we propose a strategy to immobilize more active and stable amine transaminases (ATAs) by combining protein engineering with immobilization techniques. Our approach involves the structure-guided insertion of histidine clusters (His-clusters) at flexible regions of ATA subunit interfaces, enabling immobilization on cobalt-chelated carriers. By screening multiple ATAs from various microbial sources and testing different His-clusters for each, we identified the most active and stable heterogeneous biocatalysts. Notably, the immobilized H2A variant of Chromobacterium violaceum ATA (CvATA-2HA) exhibited the highest activity per mass of biocatalyst (4 U g