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JPIFACCE-Agriculture, Food Security and Climate Change

Production of ferulic acid, 2,3 Butanediole and microbial plant biostimulants from lignocellulosic biomass by a two-step cascading process

Alternative title: Production of ferulic acid, 2,3 Butanediole and microbial plant biostimulants from lignocellulosic biomass by a two-step cascading process

Awarded: NOK 3.0 mill.

Project Number:

283972

Application Type:

Project Period:

2018 - 2021

Location:

Partner countries:

The main aim of the DEBUT project was to develop a small scale biorefinery process using novel plant biostimulants, which was performed by Romanian partners in the Norwegian-Romanian ERA-NET consortium. The biorefinery process produces versatile chemicals with application to the chemical industry as well as the health and wellness market. Natural deep eutectic solvents (NADES), are a novel group of compounds that are used in the biorefinery process. Ethaline and glyceline are two of these NADES that have been evaluated. In addition, ferulic acid that is found in plant cell walls is a highly prized chemical, an anti-oxidant that is used in skin care products to reduce the signs of aging. NIVA's role within this project was to carry out a series of ecotoxicological risk assessments on the chemical mixtures produced from the biorefinery process (NADES, Ferulic acid and combined mixture). The chemical risk assessments included a series of acute and chronic bioassays in marine and freshwater organisms. The bioassays used represented multiple species from different trophic groups including algae, crustacea and fish. Overall, ethaline was more toxic to the aquatic organisms than glyceline, with no obvious difference between marine and freshwater species. The most sensitive response was observed in the marine copepod Tisbe battagliai with LC50 concentrations (concentration causing mortality in 50% of the population) of 0.025 and 0.033% of the original concentration for ethaline and glyceline respectively. This was equivalent to an approximate 4000-fold and 3000-fold dilution of the original ethaline and glyceline solutions respectively. The results of the toxicity assessment for ferulic acid showed that of the organisms tested the marine species were approximately five times more sensitive than the freshwater species. The most sensitive responses were observed in the marine algae Skeletonema pseudocostatum and the marine copepod T. battagliai with EC/LC50 concentrations of 105 and 111 mg/L respectively. In contrast, Daphnia were the most sensitive freshwater species with an LC50 of 563 mg/L. Investigations to determine the genotoxicity of these NADES to a selection of aquatic organisms has been attempted using the comet assay. The comet assay, which provides a measure of DNA damage in plant and animal cells is being developed for use in the freshwater crustacean Daphnia sp. and in unicellular algae. Challenges in the optimisation of the assay have been overcome. These involve, for example, optimising procedures for breaking down cellular membranes of marine and freshwater algae, as well as ensuring sufficient Daphnia biomass are available in order to perform the genotoxic assessment. The genotoxicity of the NADES and ferulic acid as products of the biorefinery process will be made in the final part of the project. This will provide important information on the sub-lethal effects of the chemicals, which will be used to improve the environmental risk assessment in fresh and marine waters.

At least one scientific publication will be submitted from this project

DEBUT is an international project aimed at developing a small biorefinery process, with two steps: 1) biomass pre-treatment with natural deep eutectic solvents (NADES) and feruloyl-esterase (FAE); and 2) one pot production of a versatile chemical, 2,3 butanediol (2,3-BD), from NADES-FAE pre-treated lignocellulose biomass, by simultaneous saccarification and fermentation (SSF), performed by a plant biostimulant microbial consortia. The main biochemicals resulting from the biorefinery process are ferulic acid and 2,3-BD. Ferulic acids address mainly health and wellness related industries, whilst 2,3-BD is a versatile chemical, with a fast-growing market. The proposed biorefinery process produces also agricultural inputs such as lignin, minerals and microbial plant biostimulants. The novelty of the project is related to the use of a plant biostimulants microbial consortium for simultaneous saccarification and fermentation of lignocellulose, and to the generation of in-situ secondary DES from the ferulic acid (as HBD) and either components of the initial DES or other components (as HBA) which would provide easier separation of the extracted FA from lignocellulose and impurities. For compliance with European safety regulations selected NADES, leachates from test sites and ferulic acid extracts will be evaluated for their potential adverse effects by a battery of state-of-the art techniques, to ensure that they do not affect natural ecosystem productivity.

Funding scheme:

JPIFACCE-Agriculture, Food Security and Climate Change