Hyper-production of 13C-labeled trans-resveratrol in Vitis vinifera suspension cell culture by elicitation and in situ adsorption

Bioproduction of ¹³C-labeled trans-resveratrol in plant cell culture has attracted considerable attention with regard to potential applications for human benefit and to better understanding their absorption and in vivo metabolism in humans and animals. In the present work, two elicitors (SA and JA) and adsorbents (HP2MGL) were introduced into the Vitis vinifera cell suspension culture, and the results indicated that they could work synergistically to improve the production of trans-resveratrol (2666.7mgL^-1). Afterward, 1mM [1-¹³C]-l-phenylalanine (Phe) was added to bioproduce ¹³C-labeled trans-resveratrol with the ¹³C enrichment from 35.7% at day 5 to 20.8% at day 10. Purification of the products by several chromatographic steps was reported, and the ¹³C labeling position was verified using ¹³C NMR. Our results indicated that, in this case, the rate limiting step of production was the post-biosynthetic events rather than the biosynthesis in elicited culture. Furthermore, our results suggested the importance of simultaneous optimization of biosynthetic pathways of metabolites and their post-biosynthetic steps toward achieving commercial plant cell culture. With the help of this technique, we could expect to do industrial scaling-up in a bioreactor to produce high amounts of the non-labeled and ¹³C-labeled trans-resveratrol in the near future.