Biosynthesis of hyaluronic acid in Nicotiana tabacum plants: an efficient and safe method for large-scale production

Thus far, the industrial production of Hyaluronic Acid (HA) has focused on two main methods, extraction from animal tissues or bacterial fermentation. The main drawbacks to these systems are contamination with microbial toxins and animal pathogens, the time-consuming processes of extraction and purification, and expensive investment in scaling-up production systems. Although HA precursors are naturally produced in plants, no HA is produced in plants due to the lack of Hyaluronic Acid Synthase (HAS). In this study, the human hyaluronic acid synthase2 (hHAS2) sequence was optimized and expressed in Nicotiana tabacum using Agrobacterium- mediated transformation. HA quantitative and qualitative analysis confirmed the integration and expression of the hHAS2 in transgenic plants. The highest hHAS2 concentration of 42 ng/kg (wet weight) in transgenic lines was measured by the hHAS2 ELISA kit. The production of HA polymers in the transgenic plants was verified and quantified by the HA ELISA kit. The antioxidant activity of HA with the highest concentration of 0.17 g/kg showed a maximum activity of 52%.