Chemo‐biocatalytic one‐pot two‐step conversion of cyclic amine to lactam using whole cell monoamine oxidase

Abstract BACKGROUND Most biocatalysts currently involved in one‐pot chemoenzymatic cascades are pure enzymes, while whole cells and crude enzyme extracts remain unexplored. This work aims to develop a chemo‐biocatalytic one‐pot two‐step system involving whole cell monoamine oxidase (MAO, EC 1.4.3.4) coupled with a Cu‐based oxidative system (CuI/H 2 O 2 ) for the transformation of 1,2,3,4‐tetrahydroisoquinoline (THIQ) to 3,4‐dihydroisoquinolin‐1( 2H )‐one (DHIO). RESULTS MAO‐N variants D9 and D11 were tested as whole cell and crude lysate biocatalysts for biological oxidation. Whole Escherichia coli OverExpress C43(DE3) cells expressing MAO‐N D9 showed the best performance ( V max = 36.58 mmol L −1 h −1 , K M = 8.124 mmol L −1 , maximum specific productivity 89.3 μ mol min −1 g − 1 DCW ) and were employed in combination with CuI/H 2 O 2 in a sequential one‐pot two‐step process. The biotransformation was scaled‐up to the initial volume of 25 mL and after triple THIQ feeding, 48.2 mmol L −1 of the intermediate 3,4‐dihydroisoquinoline (DHIQ) was obtained with a yield of 71.3%. Afterwards, chemical catalysts (1 mol% CuI and 10 eq. H 2 O 2 ) were added to the biologically produced DHIQ, which was transformed to ∼30 mmol L −1 DHIO at 69.4% overall yield. CONCLUSION As MAO‐N variants have wide substrate specificity, this work broadens the portfolio of one‐pot chemoenzymatic processes employing whole cell biocatalysts, representing an alternative to using pure enzymes. © 2016 Society of Chemical Industry