Conversion Of Cellobiose Research Articles

Conversion of Cellobiose to Formic Acid as a Biomass-Derived Renewable Hydrogen Source Using Solid Base Catalysts.

Formic acid is considered a promising hydrogen carrier. Biomass-derived formic acid can be obtained by oxidative decomposition of sugars. This study explored the production of formic acid from cellobiose, a disaccharide consisting of d-glucose linked by β-glycosidic bonds using heterogeneous catalysts under mild reaction conditions. The use of alkaline earth metal oxide solid base catalysts like CaO and MgO in the presence of hydrogen peroxide could afford formic acid from cellobiose at 343 K. While CaO gave 14 % yield of formic acid, the oxide itself was converted to a harmful metal peroxide, CaO2 after the reaction. In contrast, MgO could produce formic acid without the formation of the metal peroxide. The difficulty in selectively synthesizing formic acid from cellobiose using these solid base catalysts was due to the poor conversion of cellobiose to glucose. Using a combination of solid acid and base catalysts, a high formic acid yield of 33 % was obtained under mild reaction conditions due to the quantitative hydrolysis of cellobiose to glucose by a solid acid followed by the selective decomposition of glucose to formic acid by a solid base.

Tandem catalytic conversion of glucose and cellobiose to 5-ethoxymethylfurfural with record yields

The conversion of biomass-derived carbohydrates to 5-ethoxymethylfurfural (EMF) is a fascinating reaction with 100% carbon atom utilization. Although effective conversion of 5-hydroxymethylfurfural (HMF) and fructose to EMF have been achieved, the direct conversion of glucose only gave low EMF yields. Herein, we reveal that the combinations of common Lewis acid with Brønsted acid in ethanol universally catalyze rapid etherification of glucose to intractable ethyl glucoside (EGL), as impedes the further conversion toward the desired products. The synergy of large-pore tin phosphate (SnPO) bearing strong Lewis acidic sites with acidic ionic liquid 1-(4-sulfobutyl)-3-methylimidazolium chloride (BSO3HMIMCl) enable tandem catalytic conversion of glucose, cellobiose and starch to EMF with record yields (61.7%, 61.8% and 55.7%, respectively), far exceeding previous reports. This transformation probably occurs via multiple parallel pathways, where water molecules generated by dehydration cooperate with acid and Cl- to hydrolyze EGL back to glucose, thus forming a positive feedback on EMF production.

Magnetically separable spent coffee grounds as a potential novel support for the covalent immobilization of β‐glucosidase for cellobiose hydrolysis

AbstractBACKGROUNDThe industrial‐scale application of enzymes faces obstacles due to elevated costs and difficulties in stability and reuse. In this study, magnetic spent coffee grounds, an ecotoxic waste, have been utilized successfully for the first time to immobilize β‐glucosidase to overcome these challenges.RESULTSThe spent coffee grounds were magnetized and amine‐functionalized, followed by characterization using various techniques. Under optimized conditions, forming an imine bond between the functionalized support and β‐glucosidase resulted in a 62% immobilization yield (92.81 mg g−1 enzyme loading) and 12.5 U mg−1 activity after immobilization. A relatively small kinetic change was observed in the Km value (902 to 946 μmol L−1) after immobilization, suggesting minimal hindrance by AMSCG3 on substrate access or product release. Moreover, Glu@AMSCG3 showed exceptional stability (>90% residual activity) within a pH range of 3 to 6 after 2 h of incubation at 25 °C. A residual activity of 87.94% was maintained even at 80 °C and pH 5 after 2 h of incubation compared to the free β‐glucosidase, which showed only 6.5% residual activity at the same temperature. When cellobiose was hydrolyzed using Glu@AMSCG3 under optimum conditions, 91.33% cellobiose conversion was achieved initially, and over 79% conversion was maintained for 10 reusability cycles.CONCLUSIONThe improved stability of β‐glucosidase after covalent immobilization on amine‐modified magnetically separable spent coffee grounds indicates their potential as a support matrix for application in enzyme immobilization. © 2024 Society of Chemical Industry (SCI).

Theoretical study of cellobiose conversion by supported metal catalysts

Using density functional theory and metadynamics simulations with density functional tight-binding theory, we study cellobiose hydrolysis and glucose hydrogenation with SiO2-supported Pt and Pd catalysts in hot water, relevant to green cellulose conversion. We found that cellobiose can be hydrolyzed by hydrogen atom adsorbed on metal or proton spilled-over SiO2 to form glucose. Glucose can then be hydrogenated by hydrogen atoms adsorbed at metal/water interface forming sorbitol. The reaction barriers of hydrolysis and hydrogenation at Pt/water interface are lower than at Pd/water interface, which explains an experimental finding that Pt performs better than Pd as a catalyst.

Numerical simulation of enzymatic hydrolysis in a bionic intestinal segmentation reactor

Enzymatic hydrolysis plays a critical role in the microbial conversion of lignocellulosic biomass, which has been widely performed in continuous stirred-tank reactors (CSTRs). However, low hydrolysis efficiency in CSTRs hinders the industrial application of lignocellulose to produce biofuels. A bionic intestinal segmentation reactor (BISR) was proposed for efficient enzymatic hydrolysis. In BISR, the dynamic reactor's wall with enzyme secretion led to a 2.0-fold improvement in the conversion of cellobiose as a model substrate compared to the control with a static one. Meanwhile, the reaction rate reached 0.89 mmol/l/s, which was increased by 6.7-fold. Flow fields further illustrated that secondary flow during the segmentation facilitated radial mass transfer between cellobiose and enzyme. Dimensionless secondary flow intensity Sem increased from 0.93 to 2.35 with amplitude, while decreasing from 2.86 to 2.26 with period. More importantly, the analysis of dimensionless Péclet number (Pe) and Damköhler number (Da) indicated that secondary flow could significantly enhance the hydrolysis process controlled by mass transfer.

An amino and triazole-containing metal-organic framework: cellobiose sensing and its catalytic conversion under mild conditions.

A novel metal-organic framework functionalized with amino and triazole moieties, namely Zn-TDA, has been synthesized. It represents the first example of a MOF-based sensor for highly selective sensing of cellobiose over other saccharides. Furthermore, Zn-TDA was used as a catalyst for the conversion of cellobiose to 5-hydroxymethylfurfural (5-HMF), achieving an excellent yield of 68.7% within 90 min under mild conditions.

Selective cellobiose photoreforming for simultaneous gluconic acid and syngas production in acidic conditions

Here, we demonstrate the selective cellobiose (building block of cellulose) photoreforming for gluconic acid and syngas co-production in acidic conditions by rationally designing a bifunctional polymeric carbon nitride (CN) with potassium/sulfur co-dopant. This heteroatomic doped CN photocatalyst possesses enhanced visible light absorption, higher charge separation efficiency than pristine CN. Under acidic conditions, cellobiose is not only more efficiently hydrolyzed into glucose but also promotes the syngas and gluconic acid production. Density functional theory (DFT) calculations reveal the favorable generation of •O2− during the photocatalytic reaction, which is essential for gluconic acid production. Consequently, the fine-designed photocatalyst presents excellent cellobiose conversion (>80%) and gluconic acid selectivity (>70%) together with the co-production of syngas (∼56 μmol g−1 h−1) under light illumination. The current work demonstrates the feasibility of biomass photoreforming with value-added chemicals and syngas co-production under mild condition.

Efficient and Economic Utilization of Cellobiose for Glycosylation Modification by Regulating Carbon Source Supply and Metabolic Pathway In Vivo.

Glycosylation, one of the most common and significant modifications in nature, has prompted the development of a cellobiose phosphorolysis route for glycosylation in vivo. However, the process of glycosylation is hampered by the notably low conversion rate of cellobiose. In this work, regulation of the carbon source supply by changing the ratio of glucose to cellobiose improved the conversion rate of cellobiose, resulting in enhancing the efficiency of glycosylation and the production of vitexin. Moreover, three genes (pgm, agp, and ushA) involved in the degradation of UDP-glucose were knocked out to relieve the degradation and diversion of the cellobiose phosphorolysis route. Finally, through the optimization of conversion conditions, we observed a continuous enhancement in cellobiose conversion rate and vitexin production in BL21ΔushAΔagp-TcCGT-CepA, corresponding to an increased concentration of added glucose. The maximum production of vitexin reached 2228 mg/L with the addition of 2 g/L cellobiose and 6 g/L glucose, which was 312% of that in BL21-TcCGT-CepA with the addition of 2 g/L cellobiose. The conversion rate of cellobiose in BL21ΔushAΔagp-TcCGT-CepA reached 88%, which was the highest conversion rate of cellobiose to date. Therefore, this study presents a cost-effective and efficient method to enhance the conversion rate of cellobiose during the glycosylation process.

Kinetic and thermodynamic evidences of the Diels-Alder cycloaddition and Pechmann condensation as key mechanisms of hydrochar formation during hydrothermal conversion of Lignin-Cellulose

We investigated the kinetics and thermodynamics of the Diels-Alder cycloaddition and Pechmann condensation to evaluate the significance of both mechanisms toward hydrochar formation during hydrothermal carbonization and liquefaction of biomass. We performed well-designed experiments of semi-continuous hydrothermal conversion of pure compounds representative of lignin-cellulose fractions (i.e., D-(+)-cellobiose and 1-(3,4-dimethoxyphenyl)-2-(4-hydroxy-2-methoxyphenoxy)propane-1,3-diol) and corresponding hydrochar precursors at 220–370 °C for 60 min. We developed thermodynamically-feasible multi-phase reaction pathways and specified the kinetic and thermodynamic parameters of each pathway by fitting the reversible power-law kinetic model, Arrhenius equation, and correlation between conventional definition of Gibbs free energy and the Gibbs free energy isotherm equation into experimental data of time evolution of chemical species concentration measured using GC–MS, XRD, and μGC. Reaction pathways of mixtures of pure compounds were a combination of those for individual pure compounds. We discovered that endothermic Diels-Alder cycloaddition involving dienes and dienophile was prominent in D-(+)-cellobiose conversion pathways and functioned as an intermediate reaction producing precursors for various endothermic and exothermic precipitation mechanisms. The temperature dependence of solid organics formation via this indirect route depended on the net ΔH of a set of reaction pathways after Diels-Alder cycloaddition. In contrast, we found that the Pechmann condensation involving phenols and β-ketoester/β-ketoacid was significant in the conversion pathway of 1-(3,4-dimethoxyphenyl)-2-(4-hydroxy-2-methoxyphenoxy)propane-1,3-diol and facilitated direct precipitation of organics including 4,10-dimethyl-2,7-dioxo-2,7-dihydropyrano[2,3-g]chromene-5-carbaldehyde and 4,5-dimethoxy-2,7-dioxo-6,7-dihydro-2H-benzo[g]chromene-10-carboxylic acid. The endothermic nature of Pechmann condensation preferred higher temperatures for a higher equilibrium conversion toward solid product formation. Results in this study contributed to the improved understanding of fundamentals of hydrochar formation.

Catalytic valorization of biomass carbohydrates into levulinic acid/ester by using bifunctional catalysts

Methyl levulinate (ML) and levulinic acid (LA), important platform chemicals derived from biomass carbohydrates, are potential candidates for use as fuel additives and chemicals. In this work, bifunctional solid acid catalysts were prepared by impregnation via an environmentally friendly method and used for the conversion of glucose to ML and LA. In the conversion of glucose to levulinate, the isomeric conversion of glucose into fructose mainly utilized the Lewis acid sites properties exhibited by chromium, and the dehydration of fructose to 5-hydroxymethylfurfural and its degradation to levulinate mainly exploited the Brønsted acid sites properties exhibited by A15. The yield of ML and LA was 43.1 % and 1.2 %, respectively, and glucose was completely transformed into methanol within 2.0 h at 200 °C. The excellent activity of the best catalyst may be due to the sufficient total acidity and suitable ratio of the number of Lewis acid sites to that of Brønsted acid sites. The results for five catalysis cycles proved that the catalyst has good stability and reusability. In addition, the catalyst prepared was effective for the conversion of other carbohydrates, namely, sucrose, cellobiose, and 5-hydroxymethylfurfural to ML with yields of 83.3 %, 77.7 %, and 57.4 %, respectively.

Preparation of Ga\(_{2}\)O\(_{3}\)-modified sulfated zirconia mesopore and its application on cellobiose hydrolysis

Heterogeneous catalyst plays vital role in biomass processing due to slow rate of biological and naturally pathway processes. Solid acid sulfated zirconia (SZ) is a promising catalyst with properties that can be tuned up. Sulfated zirconia was successfully modified by 2%, 5% and 10% (wt.) Ga2O3 (xGa-SZ; x = 2, 5 and 10) via template-assisted sol-gel method. The catalysts were characterized through various method (XRD, SEM-EDS and Gas Sorption analysis) and applied on hydrolysis of cellobiose, a model compound of cellulose. Diffraction pattern showed xGa-SZ formed completely tetragonal phase whereas un-promoted SZ contains mixed phase of monoclinic and tetragonal. Acidity evaluation via gravimetric method using ammonia as probe molecule indicates the Ga2O3 promoted sulfated zirconia has larger acidity. The SEM-EDS results confirmed the presence of Gallium element on the surface of promoted xGa-SZ. Gas sorption analysis shows specific surface area is improved (83 m2∙g-1 to 123 m2∙g-1) and increased pore radii (36 Å to 56 Å). The adsorption-desorption isotherm displayed pattern of meso-porosity material. At higher T and longer time, SZ yield more glucose than xGa-SZ. However, at shorter time, 2Ga-SZ and 10Ga-SZ show better hydrolysis performance. The solid acid 10Ga- SZ shows potential performance as heterogeneous catalyst for cellobiose conversion in modest conditions.

Cooperative Catalysis Mechanism of Brønsted and Lewis Acids from Al(OTf)3 with Methanol for β-Cellobiose-to-Fructose Conversion: An Experimental and Theoretical Study.

Al-containing catalysts, e.g., Al(OTf)3, show good catalytic performance toward the conversion of cellulose to fructose in methanol solution. Here, we report the catalytic isomerization and alcoholysis mechanisms for the conversion of cellobiose to fructose at the PBE0/6-311++G(d,p), aug-cc-pVTZ theoretical level, combining the relevant experimental verifications of electrospray ionization mass spectrometry (ESI-MS), high-performance liquid chromatography (HPLC), and the attenuated total reflection-infrared (ATR-IR) spectra. From the alcoholysis of Al(OTf)3 in methanol solution, the catalytically active species involves both the [CH3OH2]+ Brønsted acid and the [Al(CH3O)(OTf)(CH3OH)4]+ Lewis acid. There are two reaction pathways, i.e., one through glucose (glycosidic bond cleavage followed by isomerization, w-G) and another through cellobiulose (isomerization followed by glycosidic bond cleavage, w-L). The Lewis acid ([Al(CH3O)(OTf)(CH3OH)4]+) is responsible for the aldose-ketose tautomerization, while the Brønsted acid ([CH3OH2]+) is in charge of ring-opening, ring-closure, and glycosidic bond cleavage. For both w-G and w-L, the rate-determining steps are related to the intramolecular [1,2]-H shift between C1-C2 for the aldose-ketose tautomerization catalyzed by the [Al(CH3O)(OTf)(CH3OH)4]+ species. The Lewis acid ([Al(CH3O)(OTf)(CH3OH)4]+) exhibits higher catalytic activity toward the aldose-ketose tautomerization of glycosyl-chain-glucose to glycosyl-chain-fructose than that of chain-glucose to chain-fructose. Besides, the Brønsted acid ([CH3OH2]+) shows higher catalytic activity toward the glycosidic bond cleavage of cellobiulose than that of cellobiose. Kinetically, the w-L pathway is predominant, whereas the w-G pathway is minor. The theoretically proposed mechanism has been experimentally testified. These insights may advance on the novel design of the catalytic system toward the conversion of cellulose to fructose.

Anchoring Ru Active Sites on N−CNTs for an Improved One‐pot Cellobiose Conversion

AbstractIn this contribution, N‐doped carbon nanotubes were obtained through nitrogen plasma treatment and used as the anchoring sites for Ru catalysts in one‐pot cellobiose conversion to hexitols. XPS and HR TEM‐EDS analysis revealed a successful introduction of N on CNTs using 300 min nitrogen plasma treatment. Both the activation ability and the amount of activated H2 were improved by the N‐anchored Ru active phase, leading to a much‐enhanced hexitols yield. The hexitols yield reached 91.5% on Ru/CNT−P‐t300 catalyst, being substantially higher than those on Ru/AC (56.84%) and Ru/CNT (76.84%). DFT simulation results verified the enhanced H2 activation ability. The Eb value of the Ru4 cluster binds with (5, 5) CNT and (5, 5) N−CNT was −4.11 and −4.66 eV, respectively, suggesting that the graphitic‐N engaged as the anchoring sites for Ru via strong interaction. The research highlights a green and facile strategy to dope N element as the anchoring sites for noble metal catalyst in improving H2 activation capability for a broad hydrogenation application.

Controlling the Adsorption of β-Glucosidase onto Wrinkled SiO2 Nanoparticles To Boost the Yield of Immobilization of an Efficient Biocatalyst.

β-Glucosidase (BG) catalyzes the hydrolysis of cellobiose to glucose, a substrate for fermentation to produce the carbon-neutral fuel bioethanol. Enzyme thermal stability and reusability can be improved through immobilization onto insoluble supports. Moreover, nanoscaled matrixes allow for preserving high reaction rates. In this work, BG was physically immobilized onto wrinkled SiO2 nanoparticles (WSNs). The adsorption procedure was tuned by varying the BG:WSNs weight ratio to achieve the maximum controllability and maximize the yield of immobilization, while different times of immobilization were monitored. Results show that a BG:WSNs ratio equal to 1:6 wt/wt provides for the highest colloidal stability, whereas an immobilization time of 24 h results in the highest enzyme loading (135 mg/g of support) corresponding to 80% yield of immobilization. An enzyme corona is formed in 2 h, which gradually disappears as the protein diffuses within the pores. The adsorption into the silica structure causes little change in the protein secondary structure. Furthermore, supported enzyme exhibits a remarkable gain in thermal stability, retaining complete folding up to 90 °C. Catalytic tests assessed that immobilized BG achieves 100% cellobiose conversion. The improved adsorption protocol provides simultaneously high glucose production, enhanced yield of immobilization, and good reusability, resulting in considerable reduction of enzyme waste in the immobilization stage.

Highly Selective Photocatalytic Conversion of Glucose on Holo-Symmetrically Spherical Three-Dimensionally Ordered Macroporous Heterojunction Photonic Crystal

Highly Selective Photocatalytic Conversion of Glucose on Holo-Symmetrically Spherical Three-Dimensionally Ordered Macroporous Heterojunction Photonic Crystal

One-pot synthesis of 5-hydroxymethylfurfural from cellobiose and sucrose using niobium-modified montmorillonite catalysts

The synthesis of platform chemical 5-hydroxymethylfurfural (5-HMF) provides favorable and efficient paths for the utilization of renewable biomass resources. In this work, highly active niobium-modified montmorillonite (Nb-Mont) catalysts were synthesized by a facile and ion-exchange method and were used for the one-pot conversion of cellobiose and sucrose to 5-HMF with optimal results. 5-HMF yield of 68.6% was obtained from cellobiose in a methyl isobutyl ketone (MIBK)/saturated salt water (SSW) biphasic solvent at 170 °C after 3 h. With sucrose as substrate in DMSO at 140 °C after 2 h, 5-HMF yield was 65.4%. The presence of Lewis and Brønsted acid sites on Nb-Mont facilitate the hydrolysis of cellobiose and sucrose and play essential roles in the subsequent isomerization of glucose monomer and the dehydration of fructose monomer. A possible reaction mechanism was proposed for the transformation of cellobiose and sucrose into 5-HMF, and the isomerization process of glucose monomer was the most critical step, confirmed by density functional theory (DFT) calculations. [NbO(OH)2]+ species as active sites could effectively reduce the activation energy of the isomerization process, which accelerated the synergetic proton transfer process. Nb-Mont was recycled at least four times without a prominent loss in the catalytic activity. In this study, a promising catalytic strategy was provided for heterogeneous Nb containing catalysts used in the effective synthesis of 5-HMF from carbohydrates.

PHB production from cellobiose with Saccharomyces cerevisiae

Replacement of petrochemical-based materials with microbially produced biodegradable alternatives calls for industrially attractive fermentation processes. Lignocellulosic materials offer non-edible alternatives for cultivated sugars, but require often use of expensive sugar releasing enzymes, such as β-glucosidases. These cellulose treatment costs could be reduced if microbial production hosts could use short cellodextrins such as cellobiose directly as their substrates. In this study, we demonstrate production of poly(hydroxybutyrate) (PHB) in yeast Saccharomyces cerevisiae using cellobiose as a sole carbon source. Yeast strains expressing PHB pathway genes from Cupriavidus necator and cellodextrin transporter gene CDT-1 from Neurospora crassa were complemented either with β-glucosidase gene GH1-1 from N. crassa or with cellobiose phosphorylase gene cbp from Ruminococcus flavefaciens. These cellobiose utilization routes either with Gh1-1 or Cbp enzymes differ in energetics and dynamics. However, both routes enabled higher PHB production per consumed sugar and higher PHB accumulation % of cell dry weight (CDW) than use of glucose as a carbon source. As expected, the strains with Gh1-1 consumed cellobiose faster than the strains with Cbp, both in flask and bioreactor batch cultures. In shake flasks, higher final PHB accumulation % of CDW was reached with Cbp route (10.0 ± 0.3%) than with Gh1-1 route (8.1 ± 0.2%). However, a higher PHB accumulation was achieved in better aerated and pH-controlled bioreactors, in comparison to shake flasks, and the relative performance of strains switched. In bioreactors, notable PHB accumulation levels per CDW of 13.4 ± 0.9% and 18.5 ± 3.9% were achieved with Cbp and Gh1-1 routes, respectively. The average molecular weights of accumulated PHB were similar using both routes; approximately 500 kDa and 450 kDa for strains expressing either cbp or GH1-1 genes, respectively. The formation of PHB with high molecular weights, combined with efficient cellobiose conversion, demonstrates a highly potential solution for improving attractiveness of sustainable polymer production using microbial cells.

Rapid evolution and mechanism elucidation for efficient cellobiose-utilizing Saccharomyces cerevisiae through Synthetic Chromosome Rearrangement and Modification by LoxPsym-mediated Evolution

Lack of cellobiose utilization capability for many microorganisms results in carbon source waste in lignocellulosic biorefinery. In this study, genes for cellobiose transport and hydrolysis were introduced to Saccharomyces cerevisiae synV, a semi-synthetic yeast with an inducible SCRaMbLE (Synthetic Chromosome Rearrangement and Modification by LoxPsym-mediated Evolution) system incorporated into its chromosome V, endowing cellobiose utilization capability to this strain. Thereafter, two evolved strains with 98.1% and 79.2% improvement, respectively, in cellobiose utilization rate were obtained through induced SCRaMbLE. Further studies suggested that the enhanced cellobiose utilization capability directly correlated with copy number increases of introduced genes and some chromosome structural variations. In particular, it was experimentally demonstrated for the first time that deletion of redox stress related gene MXR1 and ATP conversion related gene ADK2 contributed to enhanced cellobiose conversion. Thereafter, the effectiveness of MXR1 and ADK2 deletions was demonstrated in artificial hydrolysate and rice straw hydrolysate, respectively.

Immobilization-Stabilization of β-Glucosidase for Implementation of Intensified Hydrolysis of Cellobiose in Continuous Flow Reactors

Cellulose saccharification to glucose is an operation of paramount importance in the bioenergy sector and the chemical and food industries, while glucose is a critical platform chemical in the integrated biorefinery. Among the cellulose degrading enzymes, β-glucosidases are responsible for cellobiose hydrolysis, the final step in cellulose saccharification, which is usually the critical bottleneck for the whole cellulose saccharification process. The design of very active and stable β-glucosidase-based biocatalysts is a key strategy to implement an efficient saccharification process. Enzyme immobilization and reaction engineering are two fundamental tools for its understanding and implementation. Here, we have designed an immobilized-stabilized solid-supported β-glucosidase based on the glyoxyl immobilization chemistry applied in porous solid particles. The biocatalyst was stable at operational temperature and highly active, which allowed us to implement 25 °C as working temperature with a catalyst productivity of 109 mmol/min/gsupport. Cellobiose degradation was implemented in discontinuous stirred tank reactors, following which a simplified kinetic model was applied to assess the process limitations due to substrate and product inhibition. Finally, the reactive process was driven in a continuous flow fixed-bed reactor, achieving reaction intensification under mild operation conditions, reaching full cellobiose conversion of 34 g/L in a reaction time span of 20 min.

Biosynthesis of fuel-grade ethanol from cellobiose by a cell-factory of non-GMO Saccharomyces cerevisiae/starch-gel-cellulase

The aim of this work was to ferment cellobiose to fuel-grade ethanol in one-step bioconversion employing a cell-factory (CF) of non-Genetically Modified Organism (non-GMO) Saccharomyces cerevisiae. The proposed technology involved the production of a freeze-dried preparation of CF of non-GMO S. cerevisiae covered by a layer of starch-gel (SG) containing cellulase enzyme. Parameters studied for one-step bioconversion of cellobiose to ethanol included temperature (33–40 °C), cellobiose concentration (70–150 g/L), and cellulase activity (60–150 FPU/g). High conversion (90%) of cellobiose in a 48 h one-step process was achieved using 70 g/L cellobiose, 150 FPU/g at 30 °C, indicating the efficiency of CF. S. cerevisiae in CF were also capable to ferment at 40 °C with the major role of enzyme activity and cellobiose concentration in fermentation. In our studies, the ethanol yield and conversion of cellobiose were found competitive, with other processes reported with S. cerevisiae as GMO and enzyme co-immobilized in alginates beads. Consequently, the results revealed a new concept for bio-ethanol production from cellobiose in one-step simultaneous-hydrolysis-fermentation (OSHF), which could be a precursor study for large-scale cellulose bioconversion for fuel-grade alcohol production.