Laboratory evolution of Pyrococcus furiosus alcohol dehydrogenase to improve the production of (2S,5S)-hexanediol at moderate temperatures

There is considerable interest in the use of enantioselective alcohol dehydrogenases for the production of enantio- and diastereomerically pure diols, which are important building blocks for pharmaceuticals, agrochemicals and fine chemicals. Due to the need for a stable alcohol dehydrogenase with activity at low-temperature process conditions (30°C) for the production of (2S,5S)-hexanediol, we have improved an alcohol dehydrogenase from the hyperthermophilic archaeon Pyrococcus furiosus (AdhA). A stable S-selective alcohol dehydrogenase with increased activity at 30°C on the substrate 2,5-hexanedione was generated by laboratory evolution on the thermostable alcohol dehydrogenase AdhA. One round of error-prone PCR and screening of ∼1,500 mutants was performed. The maximum specific activity of the best performing mutant with 2,5-hexanedione at 30°C was tenfold higher compared to the activity of the wild-type enzyme. A 3D-model of AdhA revealed that this mutant has one mutation in the well-conserved NADP(H)-binding site (R11L), and a second mutation (A180V) near the catalytic and highly conserved threonine at position 183

Novel basin modelling concept for simulating deformation from mechanical compaction using level sets

As sedimentation progresses in the formation and evolution of a depositional geologic basin, the rock strata are subject to various stresses. With increasing lithostatic pressure, compressional forces act to compact the porous rock matrix, leading to overpressure buildup, changes in the fluid pore pressure and fluid flow. In the context of petroleum systems modelling, the present study concerns the geometry changes that a compacting basin experiences subject to deposition. The purpose is to track the positions of the rock layer interfaces as compaction occurs. To handle the challenge of potentially large geometry deformations, a new modelling concept is proposed that couples the pore pressure equation with a level set method to determine the movement of lithostratigraphic interfaces. The level set method propagates an interface according to a prescribed speed. The coupling term for the pore pressure and level-set equations consists of this speed function, which is dependent on the compaction law. The two primary features of this approach are the simplicity of the grid and the flexibility of the speed function. A first evaluation of the model concept is presented based on an implementation for one spatial dimension accounting for vertical effective stress. Isothermal conditions with a constant fluid density and viscosity were assumed. The accuracy of the implemented numerical solution for the case of a single stratigraphic unit with a linear compaction law was compared to the available analytical solution [38]. The multi-layer setup and the nonlinear case were tested for plausibility