Modification of chimeric (2S,3S)-butanediol dehydrogenase based on structural information

A chimeric (2s,3s)-butanediol dehydrogenase (clbdh) was engineered to have the strict (s)-configuration specificity of the (2s,3s)-bdh (bslbdh) derived from brevibacterium saccharolyticum as well as the enzymatic stability of the (2r,3s)-bdh (kpmbdh) from klebsiella pneumonia by swapping the domains of two native bdhs. however,while clbdh possesses the stability,it lacks the specificity. in order to assist in the design a bdh having strict substrate specificity,an x-ray structural analysis of a clbdh crystal was conducted at 1.58 å. the results obtained show some readily apparent differences around the active sites of clbdh and bslbdh. based on this structural information,a novel (2s,3s)-bdh having a preferred specificity was developed by introducing a v254l mutation into clbdh. the influence of this mutation on the stability of clbdh was not evaluated. nevertheless,the technique described herein is an effective method for the production of a tailor-made bdh. © 2015 bentham science publishers.