Consolidated bioprocessing of corn cob-derived hemicellulose: engineered industrial Saccharomyces cerevisiae as efficient whole cell biocatalysts

Background Consolidated bioprocessing, which combines saccharolytic and fermentative abilities in a single microorganism, is receiving increased attention to decrease environmental and economic costs in lignocellulosic biorefineries. Nevertheless, the economic viability of lignocellulosic ethanol is also dependent of an efficient utilization of the hemicellulosic fraction, which contains xylose as a major component in concentrations that can reach up to 40% of the total biomass in hardwoods and agricultural residues. This major bottleneck is mainly due to the necessity of chemical/enzymatic treatments to hydrolyze hemicellulose into fermentable sugars and to the fact that xylose is not readily consumed by Saccharomyces cerevisiaethe most used organism for large-scale ethanol production. In this work, industrial S. cerevisiae strains, presenting robust traits such as thermotolerance and improved resistance to inhibitors, were evaluated as hosts for the cell-surface display of hemicellulolytic enzymes and optimized xylose assimilation, aiming at the development of whole-cell biocatalysts for consolidated bioprocessing of corn cob-derived hemicellulose. Results These modifications allowed the direct production of ethanol from non-detoxified hemicellulosic liquor obtained by hydrothermal pretreatment of corn cob, reaching an ethanol titer of 11.1 g/L corresponding to a yield of 0.328 g/g of potential xylose and glucose, without the need for external hydrolytic catalysts. Also, consolidated bioprocessing of pretreated corn cob was found to be more efficient for hemicellulosic ethanol production than simultaneous saccharification and fermentation with addition of commercial hemicellulases. Conclusions These results show the potential of industrial S. cerevisiae strains for the design of whole-cell biocatalysts and paves the way for the development of more efficient consolidated bioprocesses for lignocellulosic biomass valorization, further decreasing environmental and economic costs.This work has been carried out at the Biomass and Bioenergy Research Infrastructure (BBRI)-LISBOA-01-0145-FEDER-022059, supported by Operational Programme for Competitiveness and Internationalization (PORTUGAL2020), by Lisbon Portugal Regional Operational Programme (Lisboa 2020) and by North Portugal Regional Operational Programme (Norte 2020) under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) and has been supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020, the “Contrato-Programa” UIDB/04050/2020, the MIT-Portugal Program (Ph.D. Grant PD/BD/128247/2016 to Joana T. Cunha) and through Project FatVal (POCI-01-0145-FEDER-032506) and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte.info:eu-repo/semantics/publishedVersio

Applications of yeast flocculation in biotechnological processes

A review on the main aspects associated with yeast flocculation and its application in biotechnological processes is presented. This subject is addressed following three main aspects – the basics of yeast flocculation, the development of “new” flocculating yeast strains and bioreactor development. In what concerns the basics of yeast flocculation, the state of the art on the most relevant aspects of mechanism, physiology and genetics of yeast flocculation is reported. The construction of flocculating yeast strains includes not only the recombinant constitutive flocculent brewer’s yeast, but also recombinant flocculent yeast for lactose metabolisation and ethanol production. Furthermore, recent work on the heterologous β-galactosidase production using a recombinant flocculent Saccharomyces cerevisiae is considered. As bioreactors using flocculating yeast cells have particular properties, mainly associated with a high solid phase hold-up, a section dedicated to its operation is presented. Aspects such as bioreactor productivity and culture stability as well as bioreactor hydrodynamics and mass transfer properties of flocculating cell cultures are considered. Finally, the paper concludes describing some of the applications of high cell density flocculation bioreactors and discussing potential new uses of these systems.Fundação para a Ciência e a Tecnologia (FCT) – PRAXIS XXI - BD11306/97

De invloed van de verstuivercapaciteit op de energetische en ecologische prestaties van een huishoudelijke stookolieketel

peer reviewedA.I.E.- ENERGY CONSERVATION AND EMISSIONS REDUCTION IN COMBUSTION - Convention visa n° 04/45785/STE

Yeast flocculation: competition between nonspecific repulsion and specific bonding in cell adhesion.

Yeast flocculation is governed by the competition between electrostatic repulsion (nonspecific interaction) and polysaccharide-protein bonds (specific interaction). The electrical surface potential, which is mainly due to phosphodiester linkages (of the cell wall phosphomannan), maintains the cells dispersed. Polysaccharides and proteins of the cell surface can readily penetrate the potential barrier and may establish specific bonds. The specific inhibition of flocculation by various mannosyl derivatives suggested that the protein receptor binds to the group Man alpha----3 Man alpha----PO4- ----6 Man alpha----2 Man alpha... of phosphomannan. Calcium, which is required for flocculation, could act as a bridge between the negatively charged groups of phosphomannan and those of the protein receptor. The role of calcium, however, cannot be restricted only to charge neutralization because other divalent cations inhibit flocculation; our results show that cation binding is governed by strong stereochemical constraints. Studies on protein-polysaccharide interactions have shown that electrical charges may remain uncompensated at short distance, but can be stabilized by hydrogen bonds. Calcium could induce a "locked" conformation of the receptor; this conformation is the only one capable of binding phosphomannan strongly enough to make cell adhesion possible

Fluorescence study of lectinlike receptors involved in the flocculation of the yeast Saccharomyces cerevisiae.

Flocculation of some yeasts involves lectinlike receptors with two different patterns of inhibition by sugars: mannose sensitive (MS) and glucose-mannose sensitive (GMS). The visualization and quantification of these receptors were performed using neoglycoproteins fluorescent probes. Fluorescence microscopy showed a homogeneous distribution of surface receptors for the strain belonging to the MS group and a polar distribution for cells belonging to the GMS group. Affinity constants, estimated by fluorimetry, were shown to have different values (MS, 2.6 +/- 0.7 x 10(5) M-1; GMS, 2 +/- 1 x 10(6) M-1), but the number of sites was estimated to be smaller for strain NCYC 1195 which belongs to the GMS group than for strain NCYC 869 from the MS group (MS, 2.4 +/- 0.2 x 10(7) sites/cell; GMS, 3.9 +/- 0.8 x 10(6) sites/cell)

Yeast flocculation: factors affecting the measurement of flocculence.

The physical meaning of the residual absorbance of a yeast suspension after flocculation and settling has been investigated. Starting with a dispersed suspension, agitation accelerates flocculation by increasing the probability of collision between particles. As flocculation advances, agitation also breaks the flocs. A stationary state is reached when flocculence (tendency to flocculate) is counterpoised by agitation. If the intensity of agitation is maintained constant, the free cell concentration reflects the flocculence, provided the stationary state is reached. The residual absorbance, determined after settling of the flocs, is a measure of the free cell concentration and represents an adequate parameter to characterize yeast flocculence

Apports récents sur l'anthropologie des Mésolithiques et des Néolithiques mosans

Chapitre 8info:eu-repo/semantics/publishe