At the heart of bioprocesses the activity and the physiological state of microorganisms are variables still difficult to assess. Most of the information is obtained from delayed off-line measurements and remains insufficient for the development of real time control strategies to optimize the potential of micro-organisms and design high performance processes. On-line quantification of the physiological state of cells is paramount for the understanding and improvement of cell metabolism and thus to control pathways of interest. The main objective of SPECTRE is to develop an on-line system able to monitor the physiological state of microorganisms during fermentation or cell cultures. SPECTRE is partly in continuity with the work developped during the previous ANR FASST program (Programme ANR-06-BIOE-003-01-FASST : Fermentation Alcoolique d'hydrolysats lingo-cellulosiques et obtention de Souches adaptées aux Stress Technologiques). During FASST, advanced methods for the determination of yeast strain viability state were developed. In association with off-line data, on-line dielectric spectroscopy was able to track variations of cell cytoplasm conductivity and microscopy image analysis showed that cell size distribution and cell optical properties were strongly correlated with yeast cell viability. The results of the program have been positively evaluated by the ANR and ADEME expert boards. Dielectric spectroscopy (DS) has been operational for the last ten years. This technique is now routinely used in a number of cell culture and fermentation processes for the determination of biomass concentration. However, it can also give access to informations dependent on the biomass state, but has to be completed by additional techniques to access the value of biologically significant variables. The determination of total cell volume, viability, and cell size are required to calculate the membrane capacitance Cm, representative of the cell enveloppe state, and the cytoplasmic conductivity si, a marker of water and ion exchanges between cells and their environment. Off-line measurements, on samples taken during fermentation or cell cultures, give a differed access to the information provided by the DS and are not suitable for online control. The proposed SPECTRE project is based primarily on : - the study of a coupling of two innovative technologies - spatially resolved optical spectroscopy (SRS) and dielectric spectroscopy (DS) - for the online determination of cell physiological marker variables (size, membrane capacitance, intracellular conductivity...). - the implementation of associated measurement (quantitative microscopy, flow cytometry, optical density, fluorescence, ...) which will allow - the validation of the information collected by DS and SRS, and - the selection of the most relevant additional physical variables (and their associated measurement techniques) eventually able to further improve the robustness of the physiological state evaluation of the cultivated populations. The project will lead to the development of generic tools allowing the real-time control of the physiological state of microbial populations. SPECTRE connects six academic teams expert in Bioprocess Engineering and an SME, leader in the SRS domain and in the associated data analysis techniques. Each team will use cell models chosen both for their established academic and industrial interest.

Electrochemical pressure impedance spectroscopy (EPIS) shows a high sensitivity towards transport processes in cells with gaseous reactants (Grübl, Bessler et al. 2016). This novel technique is based on analyzing the dynamic current/voltage/pressure behavior by either current excitation/pressure detection or pressure excitation/voltage detection with frequencies in the range of 100 Hz to 1 mHz. For polymer electrolyte membrane fuel cells (PEMFC), EPIS is expected to allow the observation of flow and mass transport phenomena with high accuracy. These phenomena, covering gas transport in the gas diffusion layers and the catalyst support as well as evacuation of the produced water, govern the cell performance at high-current operation. The diversity of transport phenomena, their coupling with electrochemistry and temperature, and dependence on structural properties such as pore sizes and size distributions makes understanding and modeling difficult. With conventional techniques, in particular electrochemical impedance spectroscopy (EIS), transport phenomena are difficult to be distinguished from each other and their signal may be (partially) masked by charge-transfer processes. The key hypothesis governing the present proposal is that EPIS can significantly increase measurement sensitivity and accuracy of transport-related phenomena in PEMFCs. We therefore propose a combined experimental and modeling study of EPIS for PEMFCs. A single-cell setup with pressure excitation and detection will be developed and operated by CNRS – Université de Lorraine (CNRS, France). A dynamic multi-physics model will be developed and used for data analysis by Offenburg University of Applied Sciences (HSO, Germany). The primary goal of the study is the development and evaluation of EPIS for PEMFC diagnosis. In addition, the investigation is expected to bring a significant contribution on understanding, characterization and quantification of the different transport phenomena involving gases and liquid water in the cell. Finally, the use of low-cost pressure excitation and/or detection equipment such as a loudspeaker and/or pressure sensors will be tested, for the sake of significant reduction in the cost of the overall diagnosis technique. Our vision is that EPIS can become a standard diagnosis tool in everyday lab practice complementing EIS with limited add-on effort (financially and technically) but strongly enhanced output. The three-year project will consist in continuous interaction between the two partners, CNRS being in charge of the experimental part and generation of data, HSO working on model development and data interpretation. The work packages include a preparative phase for experiments and models; the development and the use of the EPIS tool covering the measurement device and the simulation tool; interpretation of the results; and evaluation and assessment of the EPIS technique.

Plant proteins, particularly from oilseed meals, are promising renewable resources for texturizing ingredients in food and cosmetic matrices. Application of these ingredients would get rid of petroleum-based products in cosmetics, and to strengthen the use of animal proteins in food, responding to major socio-economic challenges. However, they have insufficient performance compared to currently used products. The functional limitations of plant proteins currently represent a major bottleneck to their industrial development, which must be improved to reach the quality standards of animal proteins and synthetic molecules. Enzymatic transformation processes can be implemented on protein isolates to improve their functional properties. Among these processes, proteolysis and enzymatic cross-linking are the most feasible, sustainable and compatible processes for the use of plant proteins in cosmetics and food industries. Nevertheless, the understanding and control of obtaining functional protein products by these two processes are limited by the lack of knowledge of the relationships between the product characteristics and their properties. Moreover, the process implementations result from laborious and empirical experimental approaches, leading to non-optimal production ways with regard to industrial technical, economic (cost, production duration) and environmental criteria. The main objective of PROSPER project is to develop a generic methodology for obtaining tailor-made plant protein ingredients for targeted applications in food and cosmetic, by controlling enzymatic transformation processes. Its purpose is to allow (i) an improvement in the level of understanding of the relationship between product properties and functionalities; (ii) wider use of plant proteins as functional ingredients; (iii) technological innovation to improve protein functionalities and open up new fields of application. To meet this objective, an original strategy of product engineering will be applied, structured around four main work packages. Reliable analytical tools for the characterization of the products obtained and the monitoring of proteolysis and enzymatic crosslinking processes will be developed initially. Then, the methodology aims to associate the characteristics of the products obtained with functional properties of interest using supervised machine learning methods. In a third step, relationships can be established between the characteristics of the products and a kinetic monitoring parameter of the process, as a representative criterion of a targeted functionality. Modeling/simulation tools for enzymatic transformation processes, based on experimental data regressions, will be developed. Obtained models will be coupled with the established correlations to numerically explore the influence of operating conditions sets on the targeted functionalities, and thus to identify in a rational way the original production routes of a product with targeted functionality. Then, in a last step, these models will be associated with multi-criteria optimization and decision-making tools, in order to establish the optimum functioning of these transformation routes on technical and economic criteria and to analyze these routes towards environmental impacts through life cycle analysis studies.

Photoluminescence has recently burst into the world of textiles due to the large number of potential applications ranging from security to architecture and decoration. This project, aims to develop new white light generating devices based on the use of textile materials made photoluminescent by a judicious choice of quantum dots (QDs) excited by laser diodes emitting in the blue or violet range. It is based on the eco-design of a formulation associating heavy metal-free QDs with biocompatible monomers and its coating onto a textile material by an environmentally friendly photo-induced process. In this context, the key steps of the project will consist in: • Synthesizing heavy metal-free QDs with tunable optical properties in the visible range; • Developing a photo-induced synthesis of highly photoluminescent QDs/polymer nanoassemblies by adjusting the size, composition, organization/spatial distribution of QDs in the polymer film and implementing this technology in textile applications; • Preparing new photoluminesent textiles with targeted photometric characteristics when combined with violet/blue LED chips; • Establishing the feasibility of the process by manufacturing the functionalized textile in a semi-industrial line at the R&D lab of the industrial partner. The expected result is a new generation of photoluminescent textiles for building and transportation lighting applications. These systems will be able to provide cold to warm white light and polychromatic patterns under the excitation of LEDs and with higher energy efficiency than devices currently available. The LumiTex project takes advantage of the down conversion principle to produce visible light when the photoluminescent material is excited by violet/blue LEDs. For that purpose, it promotes the development of down converters-based QDs capable of satisfying all the constraints inherent to this original approach. Thus, by exploiting the intrinsic characteristics of QDs/polymer nanomaterials, this innovative strategy makes it possible to advance LED-based lighting by correcting some of its shortcomings (homogeneity of illumination, better color rendering, absence of glare, elimination of shadows, ...) while allowing substantial savings in electrical energy. This unprecedented, fast and inexpensive approach should open the door to a new generation of technical textiles; it could also be extended to other surfaces (paper, glass, plastic, wood...) to initiate a whole series of original developments.

Soils and wastes contaminated with heavy metals are prone to create major problems because of their toxicity and their management is generally expensive. But this drawback can be turn into advantage if these solid matrices contain compounds of industrial interest. However, metal concentrations are generally too low for conventional mining and metallurgical recovery. Hence, new extraction and processing technologies must be developed to ensure production of strategic metals, while preserving soil functions, and improving soil and waste quality by decreasing their toxicity. These processes would provide a range of economic, social and environmental values from materials and lands of initial low value. AGROMINE is the conception of agro-metallurgical production chains based on the culture of hyperaccumulator plants on contaminated matrices (soils, wastes) or naturally rich in metals (ultramafic soils) to produce high value metal compounds. These chains are developed for nickel (Ni) and cobalt (Co), metals of high strategic importance. They are based on a previous work devoted to the synthesis process of ammonium and nickel sulfate double salt hexahydrate (ANSH) from the biomass of Alyssum murale. They combine agromining (or phytomining) and hydrometallurgy. • Agromining is an alternative treatment for contaminated soils and wastes, and an application of phytotechnologies to exploit secondary resources. On soils naturally rich in metals it generates incomes for farmers or managers and metal removal improves soil (or matrix) quality. Here the main innovation is the production of hyperaccumulator plants on constructed agrosystems. • Hydrometallurgy produces metals with a niche strategy, seeking forms of Ni and Co of strong industrial interest. Focus is put here on Ni and Co carboxylates, which is completely innovative, but attention will still be given on Ni and Co salts for surface treatment. The AGROMINE project involves 4 research teams of Nancy (LRGP and CRPG, LIEC, LSE of Labex Ressources 21) and two SMEs (Soléo Services and Microhumus), which have a long collaboration history. It also has strong connections with joint activities between Labex Ressources 21 and ERAMET, a major French nickel mining company. ERAMET has expressed its interest for the project by providing a support letter. The consortium maintains regular contacts with the main international actors of phytomining: Albania (UAT), Québec (INRS-ETE), China (SYSU), Australia (CMLR-UQ) and the United States (USDA). Work is organized in 1 management task and 5 scientific tasks, including: 1. Characterization of matrices, including soils, sediments and sludge: new agrosystems containing metal contaminated matrices will be designed, characterized and prepared to grow hyperaccumulators; 2. Selection of hyperaccumulators and control of metal bioavailability to identify the best Ni and/or Co hyperaccumulators for each environmental condition and metal recovery; 3. Implementation of agromining at platform scale with constructed agrosystems; 4. Hydrometallurgy for metal recovery from biomass and production of high-value compounds, based on our experience on the patented synthesis of a Ni salt (ANSH), and focus on the preparation of Co salts and Ni and Co carboxylates; 5. Life Cycle Assessment and economic evaluation of the agromining chain as well as its transfer to the end-users. AGROMINE is intended to produce economic and social value from low value material and land. It is not planned to supplant conventional mining technologies. Contrary to popular belief, our field data have shown that this process makes profit: agromining on 4 000 ha producing 200 kg Ni ha-1 converted in ANSH would give an economic benefit of c.a. € 6.15 million per year. The results obtained in AGROMINE would be of great importance for the two SMEs and for the ECONICK start-up, which is currently in an incubating process in Nancy.