{A major|A significant} drawback for {practical|sensible} application of halohydrin dehalogenase HheG in biocatalysis is its rather low thermal stability and low organic solvent tolerance. We {therefore|consequently|as a result|for that reason|thus|hence} pursued a stabilization of HheG {via|by way of|through|by means of} immobilization as cross-linked enzyme crystals. {Since|Because|Given that|Considering that|Due to the fact|Considering the fact that} glutaraldehyde inactivates HheG, we introduced a cysteine residue {in the|within the|inside the} crystal interface, which enabled thiol-specific cross-linking at predefined cross-linking {sites|websites|web sites|internet sites|web-sites|web pages}. Variant HheG D114C displayed {improved|enhanced} crystallizability and yielded {stable|steady} and catalytically active CLECs {using|utilizing|making use of|employing|working with|applying} bis-maleimidoethane as cross-linker. {Effective|Efficient|Successful|Powerful|Productive|Helpful} cross-linking {at the|in the} predefined {site|website|web site|internet site|web-site|web page} {could be|might be|could possibly be|may be|may very well be} confirmed {via|by way of|through|by means of} the CLEC crystal structure. {Compared to|In comparison to|In comparison with|When compared with} soluble enzyme, the CLECs displayed {significantly|considerably|substantially|drastically} {improved|enhanced} stability and activity at {higher|greater|larger} temperatures, {lower|reduce|reduced|decrease} pH values and {in the|within the|inside the} presence of water-miscible organic solvents, which enabled their reuse {over|more than} 21 days {in the|within the|inside the} azidolysis of cyclohexene oxide. Fmoc-Lys-OH (hydrochloride) site Ribavirin web PMID:23773119
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