Engineering Cyclic Amidases for Non-natural Amino Acid Synthesis
Hydantoinases/dihydropyrimidinases are important biotechnological enzymes involved in the production of α- and β-amino acids. Their isolation from new sources with different substrate specificities, improved activity, enantioselectivity, or higher stability continues to be of great industrial interest. Here, we provide a detailed description of how to produce high quantities of the recombinant hydantoinase/dihydropyrimidinase enzyme from Sinorhizobium meliloti CECT4114 (SmeDhp). Several techniques are combined to obtain this goal, from cloning to activity measurement by HPLC.
Key wordsα- and β-Amino acids Hydantoinases/dihydropyrimidinases Cloning Overexpression Metal-affinity HPLC
This work was supported by the Spanish Ministry of Education and Science and European Regional Development Fund (ERDF) through the project BIO2007-67009, the Andalusian Regional Council of Innovation, Science and Technology through the projects CV7-02651 and TEP4691 and the European Science Foundation COST Action CM0701.
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