Efficient biosynthesis of pinosylvin from lignin-derived cinnamic acid by metabolic engineering of Escherichia coli

Abstract

Background The conversion of lignin-derived aromatic monomers into valuable chemicals has promising potential to improve the economic competitiveness of biomass biorefineries. Pinosylvin is an attractive pharmaceutical with multiple promising biological activities. Results Herein, Escherichia coli was engineered to convert the lignin-derived standard model monomer cinnamic acid into pinosylvin by introducing two novel enzymes from the wood plant: stilbene synthase from Pinus pinea (PpSTS) and 4-Coumarate-CoA ligase from Populus trichocarpa (Ptr4CL4). The expression of Ptr4CL4 drastically improved the production of pinosylvin (42.5 ± 1.1 mg/L), achieving values 15.7-fold higher than that of Ptr4CL5 (another 4-Coumarate-CoA ligase from Populus trichocarpa) in the absence of cerulenin. By adjusting the expression strategy, the optimized engineered strain produced pinosylvin at 153.7 ± 2.2 mg/L with an extremely high yield of 1.20 ± 0.02 mg/mg cinnamic acid in the presence of cerulenin, which is 83.9% ± 1.17 of the theoretical yield. This is the highest reported pinosylvin yield directly from cinnamic acid to date. Conclusion Our work highlights the feasibility of microbial production of pinosylvin from cinnamic acid and paves the way for converting lignin-related aromatics to valuable chemicals.

E-Stilbenes: General Chemical and Biological Aspects, Potential Pharmacological Activity Based on the Nrf2 Pathway

“…PIN (3,5-dihydroxy-trans-stilbene) is found in the Pinaceae family, mainly in the heartwood of Pinus sylvestris (also known as Scots pine), and can also be found in the leaves of Pinus densiflora [ 286 , 287 ]. Pine tree parts are traditionally used in East Asia to treat a variety of health conditions, including inflammation, liver toxicity, and stomach disorders.…”

Stone Pine (Pinus pinea L.) High-Added-Value Genetics: An Overview

“…Hu et al (2022) [46] successfully engineered Escherichia coli to convert the ligninderived monomer cinnamic acid into pinosylvin by introducing an enzyme from stone pine. This enzyme, stilbene synthase (PpSTS), was efficient in the biosynthesis of pinosylvin and demonstrated the potential application for the biosynthesis of products derived from cinnamic acid.…”

“…Myristic acid 14 [46] Palmitic acid 16 [28,46,47] Palmitoleic acid 16 [46] 14-methylhexadecanoic acid 17 [41] cis-9-heptadecenoic acid 17 [46] Heptadecanoic acid 17 [46] cis-9,12-linoleic acid 18 [46,47] cis-9,12,15-linolenic acid 18 [46] cis-9-oleic acid 18 [46,47] Linoleic acid 18 [41] Oleic acid 18 [41] Pinolenic acid 18 [41] Stearic acid 18 [28,46,47] Arachidic acid 20 [28,46] cis-5,11,14-dihomo-γ-linolenic acid 20 [47] cis-11,14-eicosadienoic acid 20 [46] cis-11-gondoic acid 20 [46] Sciadonic acid 20 [41] Behenic acid 22 [41] In 1997, Wolff et al [41] attempted to find taxonomic parallels between classical classification and fatty acid analysis for pine seed oils in 49 different pine species. All tested species (including stone) were correctly clustered in sects.…”

“…Additionally, several studies on enzymes involved in evolution and environmental responses [28,41,53], seed physiology [42,45], and pathogen resistance [31,32,52] can be found for stone pine. Enzyme diversity is also a topic of interest [44][45][46], actively contributing to the knowledge of the stone pine proteome. It can be argued that evaluating the proteome is not enough to understand the full genetic components behind a species.…”