3 Results
The construction of an efficient biocatalytic in vivo cascade necessitates a balanced expression of the cascade genes to avoid side product accumulation. Besides the well-characterized initiating Cyp[18, 21], CDH and CHMO have been employed for PCL monomer synthesis from cyclohexane [16], but nothing is known about respective reaction kinetics and possible inhibitions by pathway intermediates as they have been observed before, e.g., for Bayer-Villiger monooxygenases [22, 23]. Consequently, CDH and CHMO were characterized as the first step in this study to support the rational engineering of a productive 3-step cascade based on the optimized Cyp-containing whole-cell biocatalyst[18].