Structure-Properties Correlation of Cross-Linked Penicillin G Acylase Crystals

In biocatalytic processes, the use of free enzymes is often limited due to lack long-term stability and reusability. To counteract this, can be crystallized then immobilized, generating cross-linked enzyme crystals (CLECs). As mechanical activity CLECs are crucial, different penicillin G acylases (PGAs) from Gram-positive organisms have proven promising candidates for industrial production new semisynthetic antibiotics, which characterize resulting regarding their catalytic properties. The greatest hardness Young’s modulus determined by indentation with an atomic force microscope were observed Bacillus species FJAT-PGA (26 MPa/1450 MPa), followed BmPGA (Priestia megaterium PGA, 23 MPa/1170 MPa) BtPGA (Bacillus thermotolerans 11 MPa/614 MPa). addition, FJAT- showed up 20-fold higher volumetric activities compared CLECs. Correlation structural characteristics indicated that a high solvent content low number cross-linking residues might lead reduced stability. Furthermore, seems restricted small water channels severe diffusion limitations. best our knowledge, we show first time in this study entire process chain characterization diverse industrially relevant performed at microliter scale discover most important relationships