The transition to a biobased economy necessitates utilizing renewable resources as a sustainable alternative to traditional fossil fuels. Bioconversion is a way to produce many green chemicals from renewables, e.g., biopolymers like PHAs. However, fermentation and bioconversion processes mostly rely on expensive, and highly refined pure substrates. The utilization of crude fractions from biorefineries, especially herbaceous lignocellulosic feedstocks, could significantly reduce costs. This presentation shows the microbial production of PHA from such a crude stream by a wild-type thermophilic bacterium Schlegelella thermodepolymerans [1]. Specifically, it uses crude xylose-rich fractions derived from a newly developed biorefinery process for grassy biomasses (the ALACEN process). This new stepwise mild flow-through biorefinery approach for grassy lignocellulosic biomass allows the production of various fractions: a fraction containing esterified aromatics, a monomeric xylose-rich stream, a glucose fraction, and a native-like lignin residue [2]. The crude xylose-rich fraction was free of fermentation-inhibiting compounds meaning that the bacterium S.thermodepolymerans could effectively use it for the production of one type of PHA, polyhydroxybutyrate. Almost 90% of the xylose in the refined wheat straw fraction was metabolized with simultaneous production of PHA, matching 90% of the PHA production per gram of sugars, comparable to PHA yields from commercially available xylose. In addition to xylose, S. thermodepolymerans converted oligosaccharides with a xylose backbone (xylans) into fermentable xylose, and subsequently utilized the xylose as a source for PHA production. Since the xylose-rich hydrolysates from the ALACEN process also contain some oligomeric xylose and minor hemicellulose-derived sugars, optimal valorization of the C5-fractions derived from the refinery process can be obtained using S. thermodepolymerans. This opens the way for further exploration of PHA production from C5-fractions out of a variety of herbaceous lignocellulosic biomasses using the ALACEN process combined with S. thermodepolymerans. Overall, the innovative utilization of renewable resources in fermentation technology, as shown herein, makes a solid contribution to the transition to a biobased economy.[1] W. Zhou, D.I. Colpa, H. Permentier, R.A. Offringa, L. Rohrbach, G.J.W. Euverink, J. Krooneman. Insight into polyhydroxyalkanoate (PHA) production from xylose and extracellular PHA degradation by a thermophilic Schlegelella thermodepolymerans. Resources, Conservation and Recycling 194 (2023) 107006, ISSN 0921-3449, https://doi.org/10.1016/j.resconrec.2023.107006. [2] S. Bertran-Llorens, W.Zhou. M.A.Palazzo, D.I.Colpa, G.J.W.Euverink, J.Krooneman, P.J.Deuss. ALACEN: a holistic herbaceous biomass fractionation process attaining a xylose-rich stream for direct microbial conversion to bioplastics. Submitted 2023.
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tmoA and related genes encode the alpha-subunit of the hydroxylase component of the major group (subgroup 1 of subfamily 2) of bacterial multicomponent mono-oxygenase enzyme complexes involved in aerobic benzene, toluene, ethylbenzene and xylene (BTEX) degradation. A PCR-denaturing gradient gel electrophoresis (DGGE) method was developed to assess the diversity of tmoA-like gene sequences in environmental samples using a newly designed moderately degenerate primer set suitable for that purpose. In 35 BTEX-degrading bacterial strains isolated from a hydrocarbon polluted aquifer, tmoA-like genes were only detected in two o-xylene degraders and were identical to the touA gene of Pseudomonas stutzeri OX1. The diversity of tmoA-like genes was examined in DNA extracts from contaminated and non-contaminated subsurface samples at a site containing a BTEX-contaminated groundwater plume. Differences in DGGE patterns were observed between strongly contaminated, less contaminated and non-contaminated samples and between different depths, suggesting that the diversity of tmoA-like genes was determined by environmental conditions including the contamination level. Phylogenetic analysis of the protein sequences deduced from the amplified amplicons showed that the diversity of TmoA-analogues in the environment is larger than suggested from described TmoA-analogues from cultured isolates, which was translated in the DGGE patterns. Although different positions on the DGGE gel can correspond to closely related TmoA-proteins, relationships could be noticed between the position of tmoA-like amplicons in the DGGE profile and the phylogenetic position of the deduced protein sequence.
