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Self-Assembling Plant-based Hydrogels Induced by Redox Enzymes

Periodic Reporting for period 1 - SAPHIRE (Self-Assembling Plant-based Hydrogels Induced by Redox Enzymes)

Reporting period: 2020-03-18 to 2021-05-17

The SAPHIRE project was aimed at expanding Celignis expertise from analysis service provider to bioprocess development expert. The key objectives of the project are listed below:

1. Diversification of Celignis business.
2. Attract unique and rare skill set.
3. Increased visibility of the company.

In addition to the above-mentioned objectives, there were technical objectives for the project:

1. Develop novel technology for fractionation of hemicellulose from lignocellulosic biomass.
2. Develop plant-based hydrogels that can be used for cosmetic applications.
3. Complete mass balance of the process for providing the data for techno-economic analysis of the process.

The one-year project resulted in some significant business and technical outcomes which are listed below

Business Outcomes: Celignis has diversified its services from analysis service provider to bioprocess development service provider. The company has grown from 7 employees, at the project start date to 15 employees by 2022. Company has attracted 3 chemical engineers, one microbiologist, two biotechnologists and one-scale up engineer. This would not have been possible without the technical progress that has happened in SAPHIRE project.

Technical outcomes: A novel enzyme assisted hemicellulose extraction method was developed with the yields close to 70% and purities of the crude product reaching up to 80%. This purity is a great achievement because this reduces the downstream costs significantly. Prototypes of hydrogels with good emulsifying ability, thickening properties and stabilising properties were established. The mass balances and preliminary techno-economic analysis identified the enzyme cocktails required for producing the hydrogels with better performance (aiming at personal care and cosmetics industry) and the costs associated with procurement of these enzymes and their availability. This led to a multi-partner H2020 project proposal (EnXylaScope) being written by Celignis management. This proposal was successful and the project started in May 2021. EnXylaScope concerns the discovery of novel xylan-debranching enzymes and the development of their production platforms. Successful completion of that project will allow the enzyme suppliers to produce the required enzymes in bulk for low cost, a key requirement, identified in the SAPHIRE TechnoEconomic Analysis (TEA), for the development of a commercially-viable technology. SAPHIRE has taken an idea to proof of concept and from there to a multi-partner project where 13 partners are working for one main goal. This goal is to make xylan-based hydrogels to compliment the existing hydrogels from other polysaccharides in order to replace all the synthetic liquid plastics that are being used in personal care and cosmetic products.
The project involved integration and training of innovation associate (IA) to completion of the technical objectives of the project.

The IA was fully integrated in the company and training was provided at technical level, laboratory management and project management. The technical activities in the project involved chemo-enzymatic extraction of lignin, hemicellulose and cellulose fractions and their assembling into hydrogels using xylan sidechain acting enzymes and redox enzymes. The purity of the individual fractions used to form hydrogels and the final composition of the hydrogels was analysed. The purity of the fractions extracted and used is above 75% and the final composition of the hydrogels are 50% xylan, 30% cellulose, 10% lignin and 10% is a mix of inorganics and unidentified compounds. The hydrogels produced have good swelling ability and high-water holding capacity. The increased hydrophobicity of the xylan in the hydrogels has led to emulsification ability of up to 70% EI (Emulsification Index) and viscosity of up to 106 mPas-1 at 10% solids loading. Techno-economic analysis was performed to assess the process feasibility and major cost contributors were identified. Advancement in technology to reduce the process cost is currently ongoing through a multi-partner consortium project (EnXylaScope).
The results from the SAPHIRE project were disseminated and exploited through Celignis website channels and has gained attention of large industries. This has brought new project collaborations for Celignis through which advancement of the process and product to reach the industry uptake partners requirement is underway.
The project has advanced the state of the art of xylan extraction and its conversion to hydrogels. Conventionally, high alkali concentrations are used for xylan extraction and the process was almost always performed in the bleached pulp. Bleaching the pulp is energy intensive and requires toxic or expensive chemical agents. The SAPHIRE process has established a process to extract xylan from unbleached and using mild alkali treatments with the aid of enzymes. In addition to this, xylan hydrogel composites are mostly prepared using chemical cross-linking agents, while in the SAPHIRE process the hydrogel composites were achieved purely through enzymatic process.

The advancements in the technology to make the process economically viable and to bring down the process cost to <20 Euros/Kg product is underway. This improvement will help in at least partial replacement of liquid plastics used in the personal care and cosmetics sector. This will have a great societal impact, as the contaminated water bodies due to heavy use of liquid plastics can be slowly revived and future contamination can be prevented. Public can use high performance, skin- and environment-friendly products at low cost.
Emulsifying ability of hydrogels
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