Periodic Reporting for period 1 - PROSPLIGN (Prospecting natural lignin biodiversity towards unlocking value-added bioactive motifs and molecules)
Reporting period: 2024-01-01 to 2025-06-30
The global community is facing a critical need for new, sustainable, and natural ingredients for various high-value sectors, including pharmaceuticals, cosmetics, and fragrances. While terrestrial plant biodiversity represents a vast, untapped source of bioactive compounds, many of these resources remain inaccessible due to a lack of effective technologies to unlock their potential. Lignin, the second most abundant biopolymer on Earth after cellulose, is a prime example. As a by-product of industries like pulp and paper, it is often underutilised or treated as waste. The conventional methods used to process lignin often employ harsh conditions that destroy its valuable chemical structure, losing its potential for producing high-value compounds.
The PROSPLIGN project directly addresses this challenge by pioneering a new approach to bioprospecting utilising lignin as its feedstock. The central objective is to apply state-of-the-art methodologies to explore the bioactivity potential of lignin, an abundant and sustainable biopolymer. The project's four key objectives are:
1. To map lignin's natural biodiversity and procure a wide range of European lignin sources.
2. To design complementary chemical and enzymatic methods to selectively break down lignin and isolate novel bioactive small molecules.
3. To verify the bioactivity profiles of these molecules for use in pharmaceutical, cosmetics, and fragrance applications.
4. To propose innovative and sustainable production methods, confirming their environmental and economic viability.
PROSPLIGN’s reproducible and scalable pipeline for unlocking lignin's potential aims to create a new generation of bio-based, high-value products. This will contribute to diminishing pressure on natural resources by elevating lignin from an underutilised industrial by-product into a valuable raw material with broad high-tech applications, strengthening the competitiveness of the European bioeconomy. Beyond discovery, PROSPLIGN is designing sustainable production pathways for promising compounds, either directly or following derivatisation. Their viability will be assessed through Life Cycle Analysis (LCA) and Techno-Economic Assessment (TEA), ensuring scalability and alignment with EU Green Deal goals.
By combining chemical innovation, enzyme engineering, advanced data integration, and high-throughput bioassays, PROSPLIGN avoids the pitfalls of traditional bioprospecting. No animal testing, no invasive harvesting, and no speculative genomics are required – instead, the project valorises lignin as an overlooked biomass “mines” to deliver functional molecules. In doing so, PROSPLIGN addresses Europe’s key challenges of reducing reliance on imports, offering sustainable alternatives to fossil-based chemistry, and opening new value chains in bio-based industries.
The PROSPLIGN project directly addresses this challenge by pioneering a new approach to bioprospecting utilising lignin as its feedstock. The central objective is to apply state-of-the-art methodologies to explore the bioactivity potential of lignin, an abundant and sustainable biopolymer. The project's four key objectives are:
1. To map lignin's natural biodiversity and procure a wide range of European lignin sources.
2. To design complementary chemical and enzymatic methods to selectively break down lignin and isolate novel bioactive small molecules.
3. To verify the bioactivity profiles of these molecules for use in pharmaceutical, cosmetics, and fragrance applications.
4. To propose innovative and sustainable production methods, confirming their environmental and economic viability.
PROSPLIGN’s reproducible and scalable pipeline for unlocking lignin's potential aims to create a new generation of bio-based, high-value products. This will contribute to diminishing pressure on natural resources by elevating lignin from an underutilised industrial by-product into a valuable raw material with broad high-tech applications, strengthening the competitiveness of the European bioeconomy. Beyond discovery, PROSPLIGN is designing sustainable production pathways for promising compounds, either directly or following derivatisation. Their viability will be assessed through Life Cycle Analysis (LCA) and Techno-Economic Assessment (TEA), ensuring scalability and alignment with EU Green Deal goals.
By combining chemical innovation, enzyme engineering, advanced data integration, and high-throughput bioassays, PROSPLIGN avoids the pitfalls of traditional bioprospecting. No animal testing, no invasive harvesting, and no speculative genomics are required – instead, the project valorises lignin as an overlooked biomass “mines” to deliver functional molecules. In doing so, PROSPLIGN addresses Europe’s key challenges of reducing reliance on imports, offering sustainable alternatives to fossil-based chemistry, and opening new value chains in bio-based industries.
During the first 18 months, the project has made significant technical and scientific progress across all objectives.
• Lignin Sourcing and Characterisation: A comprehensive desk-based review of European lignin biodiversity has provided a foundation for understanding the relationship between lignins and their bioactivity potential. Over 30 different lignin types sourced from diverse locations, plant species, and production methods - representing an untapped wealth of chemical information for biodiscovery. A framework for lignin quality validation was completed. Further details available to the public in summary report D2.4.
• Lignin Depolymerisation: A range of chemical and enzymatic methods for lignin depolymerisation which were validated using model compounds before testing against real lignin. The chemical-based workflow generated >500 unique samples for bioassay. The enzymatic workflow generated 120 unique depolymerised lignin for bioassay studies. Multi Variate Data Analysis (MVDA) identified critical process parameters for optimal depolymerisation outcomes. Further details available to the public in summary report D3.3.
• Bioactivity Screening and Purification: The depolymerised lignin samples were tested in preliminary bioactivity screenings relevant for pharmaceutical, cosmetics, and fragrances applications. The preliminary screenings resulted in a high "hit" rate with a substantial number of samples chosen for scale-up and purification to isolate bioactive components. The purification process has made excellent initial steps by successfully separating complex “hit” mixtures into distinct fractions of molecules with verified bioactivity. The validated bioactivity of semi-purified fractions paves the way for efficient bioactivity-guided isolation of individual compounds in the second half of the project. Further details available to the public in summary reports D4.1 and D4.8.
• Lignin Sourcing and Characterisation: A comprehensive desk-based review of European lignin biodiversity has provided a foundation for understanding the relationship between lignins and their bioactivity potential. Over 30 different lignin types sourced from diverse locations, plant species, and production methods - representing an untapped wealth of chemical information for biodiscovery. A framework for lignin quality validation was completed. Further details available to the public in summary report D2.4.
• Lignin Depolymerisation: A range of chemical and enzymatic methods for lignin depolymerisation which were validated using model compounds before testing against real lignin. The chemical-based workflow generated >500 unique samples for bioassay. The enzymatic workflow generated 120 unique depolymerised lignin for bioassay studies. Multi Variate Data Analysis (MVDA) identified critical process parameters for optimal depolymerisation outcomes. Further details available to the public in summary report D3.3.
• Bioactivity Screening and Purification: The depolymerised lignin samples were tested in preliminary bioactivity screenings relevant for pharmaceutical, cosmetics, and fragrances applications. The preliminary screenings resulted in a high "hit" rate with a substantial number of samples chosen for scale-up and purification to isolate bioactive components. The purification process has made excellent initial steps by successfully separating complex “hit” mixtures into distinct fractions of molecules with verified bioactivity. The validated bioactivity of semi-purified fractions paves the way for efficient bioactivity-guided isolation of individual compounds in the second half of the project. Further details available to the public in summary reports D4.1 and D4.8.
The key advancement is the development of a reproducible and scalable pipeline for bioprospecting from lignin. The projects proven ability to source diverse lignins from all over Europe and transform them into valuable bioactive molecules lays the groundwork for unlocking other underutilised biomass resources. The project has advanced the TRLs for its key processes:
• Chemical Depolymerisation advanced to TRL 4, with the technology successfully applied to over 30 unique lignins.
• Enzymatic Depolymerisation advanced to TRL 3, with novel enzyme produced and validated on model compounds, with application to real lignin samples underway.
• Bioactive Molecules Isolation & Purification advanced to TRL 4, having simplified complex crude mixtures of lignin-derived molecules through chromatography.
A need for further uptake/success is technologies to overcome the bottleneck of isolating bioactive molecules from heterogeneous mixtures. The project is already addressing this through a dual strategy based upon prep-HPLC and a triage protocol which prioritises the fractionation of samples based upon novelty, bioactivity and isolability.
• Chemical Depolymerisation advanced to TRL 4, with the technology successfully applied to over 30 unique lignins.
• Enzymatic Depolymerisation advanced to TRL 3, with novel enzyme produced and validated on model compounds, with application to real lignin samples underway.
• Bioactive Molecules Isolation & Purification advanced to TRL 4, having simplified complex crude mixtures of lignin-derived molecules through chromatography.
A need for further uptake/success is technologies to overcome the bottleneck of isolating bioactive molecules from heterogeneous mixtures. The project is already addressing this through a dual strategy based upon prep-HPLC and a triage protocol which prioritises the fractionation of samples based upon novelty, bioactivity and isolability.