@Article{IPB-1685,
author = {Brock, A. and Brandt, W. and Dräger, B.},
title = {{The functional divergence of short-chain dehydrogenases involved in tropinone reduction}},
year = {2008},
pages = {388-401},
journal = {Plant J.},
doi = {10.1111/j.1365-313X.2008.03422.x},
volume = {54},
abstract = {Tropane alkaloids typically occur in the Solanaceae and are also found in Cochlearia officinalis , a member of the Brassicaceae. Tropinone reductases are key enzymes of tropane alkaloid metabolism. Two different tropinone reductases form one stereoisomeric product each, either tropine for esterified alkaloids or pseudotropine that is converted to calystegines. A cDNA sequence with similarity to known tropinone reductases (TR) was cloned from C. officinalis . The protein was expressed in Escherichia coli , and found to catalyze the reduction of tropinone. The enzyme is a member of the short‐chain dehydrogenase enzyme family and shows broad substrate specificity. Several synthetic ketones were accepted as substrates, with higher affinity and faster enzymatic turnover than observed for tropinone. C. officinalis  TR produced both the isomeric alcohols tropine and pseudotropine from tropinone using NADPH \+ H\+ as co‐substrate. Tropinone reductases of the Solanaceae, in contrast, are strictly stereospecific and form one tropane alcohol only. The Arabidopsis thaliana  homologue of C. officinalis  TR showed high sequence similarity, but did not reduce tropinone. A tyrosine residue was identified in the active site of C. officinalis  TR that appeared responsible for binding and orientation of tropinone. Mutagenesis of the tyrosine residue yielded an active reductase, but with complete loss of TR activity. Thus C. officinalis  TR presents an example of an enzyme with relaxed substrate specificity, like short‐chain dehydrogenases, that provides favorable preconditions for the evolution of novel functions in biosynthetic sequences.}    
}