Protein Engineering
Engineering proteins for flavonoid biosynthesis.
 

Engineering plant proteins for improved synthesis in E. coli

Yajun Yan, Amruta Bedekar and Gloria Hu

Because plant enzyme are not naturally expressed in E. coli, they may require a certain amount of designed engineering to allow for full expression within the recombinant host. For example, the synthesis of some flavonoids in plants is mediated by the membrane-bound cytochrome-P450 enzymes. Even though E. coli has proven to be a versatile heterologous host, the expression and catalysis of plant membrane-bound P450s are hindered. To utilize the easy to culture and fast growing bacterium as a P450 biocatalyst, our laboratory has engineered some of the flavonoid P450s to allow robust in vivo synthesis of oxidized flavonoids such as hydroxylated flavonols and isoflavones. This was accomplished through the development of P450 enzymes through gene fusions. [Leonard et al., 2006] Our laboratory continues to engineering plant enzymes for improved expression in E. coli and in Yeast using a variety of traditional biochemical techniques and newly discovered biological tools.

Figure: The membrane-bound cytochrome-P450 enzyme.