Immobilization of Haloalkane Dehalogenase LinB from Sphingobium japonicum UT26 for Biotechnological Applications


Bidmanova, S., Damborsky, J., Prokop, Z.




Haloalkane dehalogenases are enzymes capable of converting a broad range of aliphatic halogenated compounds to corresponding alcohols. These dehalogenase-based biotransformations are attractive for various biological processes, e.g. biocatalysis, bioremediation and detoxification, which often require protein immobilization. Different immobilization techniques, including (i) cross-linking using glutaraldehyde, dextran polyaldehyde, disuccinimidyl suberate and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, (ii) formation of cross-linked enzyme aggregates, and (iii) entrapment methods using organically modified sol-gel ORMOCER and polyvinyl alcohol particles, were systematically investigated for immobilization of selected haloalkane dehalogenase LinB from Sphingobium japonicum UT26. All tested methods and their combinations were critically compared with regard to residual enzyme activity, leaching, and their overall application advantages and limitations. The best results with the 47%, 41% and 33% retention of initial enzymatic activity were obtained by using glutaraldehyde, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, with adipic acid dihydrazide and cross-linked enzyme aggregates methodology, respectively. Following entrapment of cross-linked enzyme aggregates into polyvinyl alcohol particles, with final 25% retention of the initial enzymatic activity, secures high degree of protection for the enzyme against harsh technological conditions, and provides biocatalyst with superior mechanical properties and easy separation from the reaction media.

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Bidmanova, S., Damborsky, J., Prokop, Z., 2013: Immobilization of Haloalkane Dehalogenase LinB from Sphingobium japonicum UT26 for Biotechnological Applications. Journal of Biocatalysis & Biotransformation 2: 1-7.

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