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Far Waste in the Landes forest

The Landes forest in our region of Nouvelle Aquitaine is the biggest artificial forest in Europe. The most common tree in this forest is the maritime pine which is an important tree accounting for 25 percent of the wood production in France. This wood production or other wood work brings many co-products like bark or sawdust; these co-products are well recycled but a part of this waste still remains. This wood waste represents a huge amount of biomass that could be used to produce many things.

We decided to focus on the transformation of this biomass into some building block, the 2,5-Furandicarboxylic acid (FDCA). A building block is a chemical that possesses some functional group allowing this molecule to be converted into a wide range of useful substances.

FDCA is an interesting building block because of its numerous uses; it can be used to produce biopolymers, that one may develop bioplastics for example,[1] but it also has great advantages in medicine as an anesthetic or even in the treatment of kidney stones [2]. FDCA have been ranked as the second most interesting bio based molecule by the Department Of Energy of the US just behind four carbon diacids [3].

Our main goal is to find a new way to produce this particular building block (FDCA) in order to reduce the use of oil which is currently the main source of building block. It could significantly prevent the pollution derived from the improper oil use.

The first part of our project is already performed in the industry in an environmentally sustainable way. The process corresponding to this first part is chemically made. Our main raw material is cellulose (coming from maritime pine) that we will use for our experiments. First of all, this cellulose will be broken down into glucose by acid hydrolysis, then this glucose will be isomerized into fructose and finally this fructose will be dehydrated into Hydroxymethylfurfural (HMF)[4].

The second part of our project, which is the conversion from HMF to FDCA is also done in the industry but is quite polluting, which is why we will use molecular biology to avoid this issue.

This process is based on the insertion of three different genes into E.coli to allow it to catalyze the transformation of HMF into FDCA. HMF is a toxic molecule and studies show that some strains are resistant to it and are able to transform HMF into FDCA. We choose three genes that are HmfH from C. basilensis [6], HmfO from P. putida [7] and Aldh1 from R. ornithinolytica [8]. This transformation is done in E. coli because it is a well-known bacterium and it also has a fast growth rate, allowing overnight cultures and genetic experimental results in mere hours.

These genes will perform the transformation of HMF into FDCA in two separated pathways : one for Aldh1 and one for HmfH that can be supported by HmfO which is an aspecific enzyme.

We will try many constructions with those genes to understand which one will demonstrate the best yield for the FDCA production. We will also try to improve these constructions, the proteins or even the strains in order to obtain a better yield, closer to the one obtained by chemical processes, but in a much more sustainable way.

References :

1. Corbion : FDCA for PEF https://www.corbion.com/bioplastics/products/fdca-for-pef
2. Lewkowski J et al., “Synthesis, chemistry and applications of 5-hydroxymethyl-furfural and its derivatives”. 2001, ARKIVOC 2001 ; pp. 17-54
3. Werpy T., Petersen G., et al., “Top Value Added Chemicals from Biomass Volume I — Results of Screening for Potential Candidates from Sugars and Synthesis Gas Top Value Added Chemicals From Biomass Volume I : Results of Screening for Potential Candidates” Other Inf. PBD 1 Aug 2004 Medium: ED; Size: 76 pp. pages (2004). DOI : 10.2172/15008859
4. DESPAX-MACHEFEL S., et al., “Développement de méthodologies de synthèse d’hydroxymethylfurfural a partir de biomasse lignocellulosique” , 2013
5. Liu J1, Tang Y., Wu K., Bi C., Cui Q., et al., “Conversion of fructose into 5-hydroxymethylfurfural (HMF) and its derivatives promoted by inorganic salt in alcohol”, Carbohydr Res, 2012, 10.1016/j.carres.2011.12.006
6. Koopman F., Wierckx N., de Winde JH., Ruijssenaars HJ., et al., “Identification and characterization of the furfural and 5-(hydroxymethyl)furfural degradation pathways of Cupriavidus basilensis HMF14”
7. “Conversion and assimilation of furfural and 5-(hydroxymethyl)furfural by Pseudomonas putida KT2440”, T. Michael et al, Juin 2017, Metabolic Engineering Communications https://doi.org/10.1016/j.meteno.2017.02.001
8. Yuana H., et al.,, “Improved production of 2,5-furandicarboxylic acid by overexpression of 5-hydroxymethylfurfural oxidase and 5-hydroxymethylfurfural/furfural oxidoreductase in Raoultella ornithinolytica BF60” , Bioresource Technologies, janvier 2018 https://doi.org/10.1016/j.biortech.2017.08.166