The aim of the IBOS project is synthesizing new enantiopure nitrogen heterocycles using biosynthetic routes. These molecules will be synthesized with optimized enzymes;oxidases, aldolases/aminotransferases or nitrile-converting enzymes. Preferably simple carbon molecules such as aldehydes and ketones will be used as substrates. I mainly focus on aldolases and , if possible, aminotranferases
[Introduction]:
Aldolases are enzymes capable of enlarging the carbon skeleton of molecules, by coupling an aldehyde to a keton. In nature different types of aldolases are used in both synthesis of a carbon backbone and in degradation of (sugar) molecules. Dihydrodipicolinate synthase (DHDPS) and 2-keto-3-deoxygluconate (KDG) aldolase are aldolases I am interested in and I ll use them in my research. In this project enzymes of thermophilic archaea and bacteria will be used, as the enzymes should preferably have a long shelf-life and high stability.
[Action]:
In blasts, using the different aldolase sequences in hyperthermophilic bacteria and archaea, many hypothetical aldolase genes were found. Currently some of them are cloned and expressed in an E. coli expression system. Further in the project the enzymes will be optimized towards unnatural substrates using directed evolution. |
