Enzyme Cascades and Synthetic Biology Routes to Non-Natural Alkaloids

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Plants and medicinal compounds from plants have been used for centuries to treat pain, infections, asthma, cancer and many of these plant natural products are used in modern medicine, for example: the analgesics morphine and codeine; anti-malarial quinine for malaria; vinblastine and homoharringtonine for cancer treatment, and ephedrine for asthma. There are over 14,000 natural compounds of the alkaloid family and many possess biological activities. Recently, there has been intensive interest in producing some of these natural compounds using microbes, rather than isolating them from plants in parts of the world that are difficult to access. Synthetic biology approaches to achieve this are proving to be extremely powerful and recent articles have highlighted the possibility of producing morphine from yeast.In previous work we have investigated some of the early steps and enzymes that use amino acids and perform transformations towards a key intermediate in the pathways that plants use to synthesise the benzylisoquinoline alkaloids, a family of over 2500 alkaloids. We now aim to use these enzymes, together with other enzymes not used in the plant pathway, and new enzymes that have been little explored in biocatalytic applications. The enzymes will be used in a laboratory bacterium to generate a wide range of structurally diverse compounds that are not found in nature. Compounds will screened for antibacterial properties. Enzymes will be used in in vitro cascades to generate three, four, five, or more multiple enzyme pathways, and also by establishing the pathways in bacteria we will be able to generate new compounds in a reproducible way using a renewable resource.