Search

Cytochrome P450 monooxygenases are highly desired biocatalysts owing to their ability to catalyse a wide variety of chemically challenging C-H activation reactions. The CYP102A subfamily of enzymes are natural catalytically self-sufficient proteins consisting of a haem and FMN-FAD reductase domain fused in a single-component system. They catalyse the oxygenation of saturated and unsaturated fatty acids to produce primarily ω-1, ω-2 and ω-3 hydroxy acids. These monooxygenases have potential applications in biotechnology; however, their substrate range is still limited and there is a continued need to add diversity to this class of biocatalysts. Herein, we present the characterisation of two new members of this class of enzymes, CYP102A25 (BMar) from Bacillus marmarensis and CYP102A26 (PHal) from Pontibacillus halophilus, both of which express readily in a recombinant bacterial host. BMar exhibits the highest activity toward myristic acid and shows moderate activity towards unsaturated fatty acids. PHal exhibits broader activity towards mid-chain-saturated (C14 -C18 ) and unsaturated fatty acids. Furthermore, PHal shows good regioselectivity for the hydroxylation of myristic acid, targeting the ω-2 position for C-H activation.

Small-scale spill detection and removal represents a fundamental concern for a wide variety of industries such as food production plants. Often these spills are detected and cleaned manually resulting in slower operations and less productivity. In this design project, a robotic solution, including detection of the spill and removal of it, is presented to maintain high productivity and operations standards in a given organization. The design consists of a plastic structure including acrylic and polylactide (PLA), a vacuum motor, spill detection electronics using image processing and control apparatus. For initial testing purposes, the robotic attachment was mounted to iRobot Create 2 as a movement platform. Testing methods consisted of rounds of spill cleaning for a variety of materials including oil and syrup. As a result of those tests, the design can be scaled to different industries such as Oil and Gas, Energy, Supermarket and Warehouses. Results show improved spill detection and removal compared to manual methods.

Cytochrome P450 monooxygenases are able to catalyse a range of synthetically challenging reactions ranging from hydroxylation and demethylation to sulfoxidation and epoxidation. As such they have great potential for biocatalytic applications but are underutilised due to often-poor expression, stability and solubility in recombinant bacterial hosts. The use of self-sufficient P450 s with fused haem and reductase domains has already contributed heavily to improving catalytic efficiency and simplifying an otherwise more complex multi-component system of P450 and redox partners. Herein, we present a new addition to the class VII family with the cloning, sequencing and characterisation of the self-sufficient CYP116B62 Hal1 from Halomonas sp. NCIMB 172, the genome of which has not yet been sequenced. Hal1 exhibits high levels of expression in a recombinant E. coli host and can be utilised from cell lysate or used in purified form. Hal1 favours NADPH as electron donor and displays a diverse range of activities including hydroxylation, demethylation and sulfoxidation. These properties make Hal1 suitable for future biocatalytic applications or as a template for optimisation through engineering.

The main purpose of Life Cycle Assessment (LCA) to date has been to evaluate life cycle impacts of different design solutions and materials for a car, its sub-systems and components. Considerable number of publications are available on LCA of automotive components. This research aims to extend the LCA approach by evaluating and comparing the effects of mass reduction of passenger seats for different vehicle models in order to provide strategic support for decision making in the development process and to validate the environmental benefits of design alternatives under investigation. For this purpose, the paper presents a comprehensive LCA of passenger seats with detailed consideration of alternative scenarios for the use phase for different vehicle models.