Researchers in the field of metabolic engineering aim to develop processes to produce
useful chemicals, sustainably and responsibly, using biotechnology. These processes are
often designed to replace products derived from fossil fuels, which are unsustainable and
contribute to climate change, or plant-based products, which compete with food
production for scarce land and are subject to supply uncertainty due to weather, crop
disease, and climate change. Here, we present two research projects in metabolic
engineering. First, we demonstrate microbial production of the red food dye betanin by
engineering the betalain biosynthesis pathway into yeast. Betanin is currently
manufactured through extraction from red beets specifically grown for dye production.
We achieved betanin production levels of 17 mg/L, which is equivalent to the amount of
betanin found in 10 g/L of beet extract. With further production improvements, this
bioprocess may become cost-competitive with agricultural production and is likely to
lead to a purer product. We also demonstrate the synthesis of a suite of non-natural
betalain dyes achieved through feeding of diverse amines to a yeast production host,
including several which have never been reported. In the second research project, we
discover that an enzyme that limits production levels of a drug family is toxic to the yeast
production host. This enzyme, norcoclaurine synthase, is critical to the production of
benzylisoquinoline alkaloids, an important family of medicines that are extracted from
plants like the opium poppy. We devised a novel subcellular compartmentalization
strategy, sequestering norcoclaurine synthase in the peroxisome to alleviate cytotoxicity
while maintaining access to the enzyme’s substrates. By targeting norcoclaurine synthase
for organellar compartmentalization, we achieved improved cell growth, final titer, and
culture productivity. These projects highlight the potential of engineering complex plant
pathways into microbial hosts for economical and sustainable chemical production.