Construction of a tailor-made L (2S,3S)-butanediol dehydrogenase by exchanging domains between native structural analogs

The development of a stable l-bdh chimera was attempted by exchanging whole domains between two native structural analogs,l-bdh and meso-bdh,because the s-configuration specificity of l-bdh is valuable from the standpoint of its application but its activity is unstable,whereas meso-bdh is stable. the domain chimeras obtained indicated that the leaf-like structures constituting three domains were likely to be mainly associated with chiral recognition,and the fourth domain,the basic domain,is likely to be mainly associated with enzyme stability. a combination of the leaf domains of l-bdh and the basic domain of meso-bdh attained a sufficient level of practical use as an artificial l-bdh chimera,because the resulting enzyme had both stability and s-configuration specificity. however,the levels of stability and specificity were slightly lower than those of the respective enzymes from which they were derived. © 2011 bentham science publishers ltd.