Abstract: The present invention pertains to the use of engineered variants of enzyme CYP102A, also known as P450-BM3, for cyclopropanation of olefins containing electron-withdrawing groups. One exemplary enzyme variant, referred to as BM3-HStar, contains five mutations away from wild-type P450-BM3, and demonstrates high activity towards cyclopropanation of olefinic substrates using ethyldiazoacetate (EDA) and other carbene transfer reagents. Products of these reactions are potential precursors of levomilnacipran derivatives, a class of compounds that have been shown to be selective inhibitors of monoamine transporters. In addition, cyclopropanation reactions with the P450-BM3 enzyme variants of the invention can be conducted in whole cells expressing the enzyme variants and can proceed under aerobic conditions.

Abstract: Pilot data and data-bearer data are formed to be mutually orthogonal and in each other's null-space. Data to be transmitted is first modulated with the data-bearer matrix and then pilot data is added thereto. The pilot data may be added to each modulated symbol, thereby increasing significantly the density of pilot information available at the receiver for channel state estimation. When pilot data is added across an entire transmitted block of data, data detection performance is improved for even fast fading Rayleigh fading channels.