Rational metabolic-flow switching for the production of exogenous secondary metabolites in bamboo suspension cells

Abstract

The synthetic biology-driven production of high-value plant secondary metabolites in microbial hosts has attracted extensive attention despite various challenges, including correct protein expression and limited supplies of starting materials. In contrast, plant cell cultures are rarely used for this purpose owing to their slow proliferation rates and laborious transformation processes. Here, we propose a “rational metabolic-flow switching” strategy to efficiently produce exogenous secondary metabolites using suspension-cultured bamboo (Phyllostachys nigra; Pn) cells as model production hosts. The Pn cells biosynthesise hydroxycinnamic acid amides (HCAAs) of putrescine as major secondary metabolites, which indicates that the phenylpropanoid and polyamine biosynthetic pathways are highly active and that the Pn cells may produce alternative secondary metabolites derived from those pathways. Stable transformants of Pn cells expressing agmatine coumaroyltransferase of barley (Hordeum vulgare) were generated with the expectation of metabolic-flow switching from HCAAs of putrescine to those of agmatine. In the recombinant Pn cells, the levels of HCAAs of putrescine decreased and the HCAAs of agmatine were produced instead. The production titre of the major product, p-coumaroylagmatine, reached approximately 360 mg/L, providing a proof-of-concept for the usefulness of “rational metabolic-flow switching” in synthetic biology using plant cell hosts.