Abstract
Phloroglucinol (1,3,5-trihydroxybenzene; PG) and its derivatives are phenolic compounds that are used for various industrial applications. Current methods to synthesize PG are not sustainable due to the requirement for carbon-based precursors and co-production of toxic byproducts. Here, we describe a more sustainable production of PG using plants expressing a native bacterial or a codon-optimized synthetic PhlD targeted to either the cytosol or chloroplasts. Transgenic lines were analyzed for the production of PG using gas and liquid chromatography coupled to mass spectroscopy. Phloroglucinol was produced in all transgenic lines and the line with the highest PhlD transcript level showed the most accumulation of PG. Over 80% of the produced PG was glycosylated to phlorin. Arabidopsis leaves have the machinery to glycosylate PG to form phlorin, which can be hydrolyzed enzymatically to produce PG. Furthermore, the metabolic profile of plants with PhlD in either the cytosol or chloroplasts was altered. Our results provide evidence that plants can be engineered to produce PG using a bacterial gene. Phytoproduction of PG using a bacterial gene paves the way for further genetic manipulations to enhance the level of PG with implications for the commercial production of this important platform chemical in plants.
Highlights
Molecular farming is used to produce valuable products in plants that are important for human and veterinary medicine and for many industrial applications[1,2,3]
Metabolic engineering of plants using bacterial genes for production of commercially important platform chemicals, polymers and non-native proteins has been used for many years[2,5,7,8,45,46,47]
We report the production of PG, a platform chemical, and its glycosylated form phlorin in Arabidopsis by introducing a bacterial gene PhlD
Summary
Molecular farming is used to produce valuable products in plants that are important for human and veterinary medicine and for many industrial applications[1,2,3]. Phloroglucinol (PG) is the simplest member of the phloroglucinols family of organic compounds consisting of more than 700 naturally occurring derivatives that exhibit a wide array of useful biological activities[9]. Phloroglucinol and its derivatives have been used in pharmaceuticals, the dyeing industry, plant tissue culture and as a precursor for manufacturing of an energetic material called 1,3,5-triamino-2,4,6 trinitrobenzene (TATB)[4,11,13]. In the fruit juice industry phlorin is widely used as an orange peel marker for juice quality[12]. It has antimicrobial activity against oral bacteria[14]. Synthesis of TATB from PG would eliminate the waste streams and red water waste associated with its chemical synthesis
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