Cellulosic Biomass Research Articles

Functionalized magnetic nanoparticles for Cellic Ctec2 cellulase immobilization: Allowing reusability of enzyme in the conversion of cellulosic biomass

The method of enzyme immobilization can ameliorate the overall stability and restoration of enzymes, hence facilitating their broader application in several sectors. This investigation utilized cellulase as a hydrolytic enzyme. In order to enhance the stability and performance of the cellulase enzyme, the research employed immobilization technology to secure the Cellic Ctec2 cellulase to the synthesized Cs@Fe3O4 nanocomposites. Fe3O4 nanoparticles (NPs) were coated with chitosan obtained from co-precipitation method that served as enzyme carrier. The NPs (Cs@ Fe3O4) were observed under XRD; VSM (vibrating-sample magnetometer) shows saturation magnetizations (Ms), UV–vis, field emission scanning electron microscopy (FESEM), and Fourier transform infrared spectroscopy (FT-IR). Response surface approach was applied to optimize the conditions for immobilization of cellulase. The optimum immobilization of cellulase reaches to 99.1% of loading efficiency and 69.7% of recovery activity with 2.5% of glutaraldehyde concentration. Furthermore, under ideal circumstances the immobilized enzyme’s thermostability, pH stability, temperature tolerance and reusability, were studied with respect to free cellulase. Higher relative activity of cellulase enzyme was observed at pH 5 with 50 °C temperature than free enzyme. One percent CMC hydrolysis is considered for reusability of free and immobilized enzyme and releases 222 mg glucose/g substrate at 24 h, showing great quiescence in cellulosic biomass conversion. Immobilized cellulase demonstrated high reusability by retaining almost 61.2% up to the 5th cycles and 51.2% of activity-maintained 10th cycle of hydrolysis. Reusability of cellulase enzyme can attain a gradual decrease in relative activity as number of repeats of the cycle increases to 10 during hydrolysis and increases in glucose yield after hydrolysis.

Productivity and quality of paulownia wood in the Right Bank Forest Steppe of Ukraine

Purpose. To determine the effect of application of fertilizer, cryoprotectant and foliar feeding on the formation of yield and quality of paulownia. Methods. The research was conducted in 2021–2023 at the experimental plot of the Bila Tserkva National Agrarian University located in the forest plantations in the Right Bank Forest Steppe of Ukraine (Kyiv region). Design of the experiment: factor A – fertilization: 1) without fertilizers, 2) application of organic fertilizer Vermykompost before planting (400 kg/ha); factor B – application of cryoprotectant: 1) without cryoprotectant, 2) cryoprotectant Mars-EL (0.5 l/ha) applied at the beginning of leaf growth; factor B – foliar application of fertilizers: 1) no treatment, 2) Quantum-AmiNoFrost (1.5 l/ha), 3) SmartGrow Recovery (2.0 l/ha). Results. At the end of the first, second and third years of paulownia cultivation, the average diameter of the trunk was 8.8; 12.2 and 17.3 cm, respectively. The application of organic fertilizer contributed to the average annual increase in this indicator by 3.0, 1.5, and 1.7 cm, and application of cryoprotector – by 0.3, 0.3, and 0.4 cm per year, respectively. The highest values of the trunk diameter in the experiment were obtained under combined application of Vermykompost, cryoprotectant and foliar fertilizer Quantum-AmiNoFrost or SmartGrow Recovery: in the first year – 10.5–10.6 cm, in the second – 13.1–13.2 cm, and in the third year – 18.4 cm. During the first, second and third years of vegetation, dry matter content of paulownia biomass per plant averaged to 3.0, 12.9, and 26.7 kg, respectively. At the same time, the application of fertilizer provided an increase in the dry matter content of 0.7, 1.7, and 3.8 kg per plant, while application of cryoprotectant provided 0.3, 0.6 and 1.2 kg per plant, respectively. Similarly to the diameter of the trunk, dry matter accumulation per plant was the highest under combined action of all factors of the experiment: in the first year – 3.58–3.60 kg, in the second – 14.1–14.2 kg, and in the third – 29.4 kg. Conclusions. The yield of dry matter of paulownia in the first year of cultivation averaged 1.88 t/ha. Application of fertilizer provided dry matter increase of 0.45 t/ha, and cryoprotectant – 0.19 t/ha in the first year; 8.04 t/ha in the second year, with increases by factors of 0.45 and 0.19 t/ha, respectively, and in the third year – 16.68 t/ha, with increases by factors of 2.40 and 0.73 t/ha, respectively. In all years of the experiment, the highest dry matter yield was recorded in the combined treatment with the use of organic fertilizer, cryoprotectant and foliar fertilization: in the first year, dry matter yield was 2.24–2.25 t/ha, in the second 8.80–8.90 t/ha, and in the third year 18.40 t/ha. The average content of cellulose in paulownia biomass over the years of research was 43.8% and lignin – 20.3%. The use of organic fertilizer provided an increase in these indicators by 1.05 and 0.29%, respectively, and cryoprotectant – by 0.41 and 0.24%. The highest content of cellulose (44.5%) and lignin (20.7%) were noted for the combined action of all three factors of the experiment. As to the ash content, the average indicator for the experiment was 1.1%, and the deviations of his parameter did not depend on the influence of the experimental factors (the difference was within the experimental error).

Production of nanocellulose from lignocellulosic biomass and its potential applications: A review

<p>Lignocellulosic biomass is a complex natural polymer primarily composed of cellulose, hemicellulose, lignin, and various other chemical components. The cellulose in the lignocellulosic biomass can depolymerize into a nano-dimension biomaterial called nanocellulose, which possesses unique features with potential application in various fields. Nanocellulose production from lignocellulosic biomass has become the subject of extensive research in the last few decades in the fields of material sciences and biomedical engineering and has attracted the attention of scientists and technologists worldwide. This production faces many challenges in utilizing the cellulose from lignocellulosic biomasses and subsequent processing for their conversion into nanocellulosic materials and their further applications in various fields of science and technology. This current review not only focuses on the production of nanocellulose from lignocellulosic biomass through different production methods but also discusses various sources, types, properties, and their applications in material science and biomedical engineering. This research review certainly shows that in the future, nanocellulose has great potential to be used as a renewable source in the field of sustainable materials and nanocomposites.</p>

A novel Ruminiclostridium thermocellum cellulase system enhances cellulosic saccharification by elimination of cellobiose feedback inhibition

The Ruminiclostridium thermocellum cellulosome system is one of the most efficient cellulase systems in nature. The main hydrolysis product of cellulosic saccharification by R. thermocellum was cellobiose, which served as a strong feedback inhibitor to the cellulosome. In this study, R. thermocellum M3, which directly hydrolyzes cellulosic biomass into monosaccharides, was used to degrade cellobiose to investigate the kinetic characteristics of cellobiose degradation. Moreover, the transcriptomic characteristics of strain M3 under cellobiose and Avicel culture conditions were compared. The results indicated that strain M3 was able to grow and hydrolyze cellobiose under cellobiose concentrations higher than 20 g/L. The best growth performance and glucose production were obtained under 12 g/L cellobiose. Compared with that of Avicel, cellobiose had greater β-glucosidase activity under cellobiose culture, with a maximum glucose yield of 767.97±19.13 mg/g cellobiose. The transcription results indicated that R. thermocellum M3 resisted the feedback inhibition of cellobiose by regulating genes encoding β-glucosidase, cellobiose phosphorylase and ABC transporters. The high glucose yield of strain M3 was closely related to the expression of the bglX gene. In addition, the gene related to the chemotactic pathway of strain M3 also increased its perception and adaptation to cellobiose.

Effect of surfactant on pseudo-lignin formation

In this study, a nonionic surfactant (JFC-M) was used as an additive for hydrothermal pretreatment of crushed poplar wood. The pseudo-lignin extracted from holocellulose after hydrothermal pretreatment was characterized, and the composition of liquid and solid fractions obtained after pretreatment at different experimental conditions was analyzed. The results showed that the addition of JFC-M surfactant accelerated the dissolution of biomass cellulose and effectively inhibited the production of pseudo-lignin in hydrothermal processes, under the same hydrothermal pretreatment conditions. The pseudo-lignin yield for the control group was 14.2% (no JFC surfactant added), whereas when the JFC-M concentration was 2%, the pseudo-lignin yield was 9.8%.

Facile Separation of Acetic Acid from 1-Ethyl-3-methylimidazolium Acetate Ionic Liquid with the Aid of a Protic Solvent.

1-Ethyl-3-methylimidazolium acetate (EmimAc), an excellent solvent for cellulosic biomass, is expected to be utilized in chemical conversion, such as in biomass acetylation with acetic anhydride. The corresponding carboxylic acid, acetic acid (AcH), is quantitatively generated as a byproduct and should be separated from EmimAc for recycling. However, the strong interaction between EmimAc and AcH makes their separation difficult under moderate conditions. This study examined the efficacy of protic solvents in distillation and extraction to weaken this interaction through solvation or hydrogen-bonding interactions. The separation efficiency of AcH from EmimAc via distillation increased as the boiling point of the protic solvent increased. Water addition was more effective than the addition of alcohols with boiling points similar to those of water such as 1-propanol and 2-butanol. Furthermore, the favorable effect of water addition on the extraction of AcH was confirmed using common organic solvents, such as diisopropyl ether, diethyl ether, and ethyl acetate. The partition coefficient (α) of AcH between the aqueous and organic phases increased with an increasing dielectric constant of the organic solvent, whereas the α value of EmimAc decreased. Repeated treatments in both distillation and extraction facilitated the complete separation of AcH from EmimAc.

Novel Technologies for Butyric Acid Fermentation: Use of Cellulosic Biomass, Rapid Bioreactor, and Efficient Product Recovery

Butyric acid, a four-carbon fatty acid, is an important industrial chemical and feedstock. To produce this chemical, a control fermentation was run with a 126.5 g.L−1 glucose concentration in the feed medium. In this medium, the strain produced 44.8 g.L−1 total acid with a productivity of 0.23 g.L−1h−1 and a yield of 0.41 g.g−1. The strain (Clostridium tyrobutyricum ATCC 25755) was also able to utilize glucose and xylose simultaneously with similar fermentation performance. The culture was also used to produce butyric acid from wheat straw hydrolysate (WSH) employing a hot water pretreatment. In a batch system, the strain resulted in a productivity and yield of 0.27 g.L−1h−1 and 0.44 g.g−1, respectively, which was an improvement over the use of glucose or xylose alone or mixtures of both. To improve reactor productivity, a membrane cell recycle bioreactor was used which resulted in a productivity of 1.89 g.L−1h−1. This productivity was 822% of that achieved in the glucose or xylose batch fermentation. Furthermore, a butyric acid recovery method was developed using XAD-4 adsorbent resin. In this system, up to 206.1 g.L−1 of butyric acid was used in the feed and, as a result of the quick adsorption, the residual butyric acid concentration was 29.5 g.L−1. In this experiment, the rate of acid removal of 1059.4 g.L−1h−1 was achieved.

Comparative Study of the Convertibility of Pretreated Miscanthus Straw Using Enzyme Preparations Produced by Different Recombinant Strains of Penicillium verruculosum

Non-edible cellulosic biomass from perennial herbaceous plants is a promising and abundant feedstock for replacing slow-growing woody plants used in biotechnological applications. Herbaceous plant biomass, as other types of plant biomass, requires pretreatment before biochemical conversion. In this study, miscanthus straw was pretreated using different methods and subjected to enzymatic hydrolysis with Penicillium verruculosum enzyme complexes under laboratory conditions. The convertibility after enzymatic hydrolysis varied from 15% to 66%, depending on the pretreatment method. Dilute alkaline pretreatment showed the highest convertibility compared to other methods, reaching up to 66%. The efficiency of dilute acid pretreatment was relatively low compared to other methods. The maximum convertibility was 37% for sulfuric acid pretreatment (the least efficient) and 51% for nitric acid. Convertibility was almost equal with 43% for white liquor and 46% for hot water. The glucose-to-xylose ratio was 4.7:1 for dilute alkaline pretreatment and 11–13:1 for white liquor. Both sulfuric and nitric acid resulted in a low xylose content in the enzymatic hydrolysates. Low-xylose hydrolysates with less than 2% of the glucose amount can be produced by hot water pretreatment. Preparation C, enriched with endoglucanase I from T. reesei and endoglucanase II from P. verruculosum, was found to be the most effective of the different enzyme preparations (EPs) tested.

Feasibility of Energy Self-Sufficiency by Biomass Production in A Brazilian State

Objective: This study aims to analyze the potential of using industrial black liquor or black lye residue from the eucalyptus pulp and paper production process as an alternative source of energy, in order to alleviate the current energy crisis. Theoretical framework: The study is based on literature regarding energy generation from biomass, focusing on the utilization of black liquor residue in thermoelectric power generation. Therefore, the thermoelectric generation from black liquor or black lye, cellulose biomass residue as an alternative source in energy generation, characterizes an important electrical feedback path in pulp production processes, also generating surplus energy for the supply network, relief of demand in the face of the current energy crisis. Method: The methodology employed in this study entails applied research, that investigates energy generation from black liquor in integrated plants of eucalyptus forest biomass, in reforestation areas for pulp production. Results and conclusion: The results indicate that energy generation from black liquor is a viable alternative to mitigate the current energy crisis, contributing to regional sustainable development. Projections suggest that by 2024, 85% of bioenergy can will be generated from this source in in the micro-region of Brazil. Therefore, adequate planning brings great expectations for regional sustainable development. Implications of the research: The research contributes to advancing scientific knowledge in the field of energy generation from biomass, demonstrating the potential of black liquor residue as an alternative energy source. Originality/value: The research contributes to advancing scientific knowledge in the field of energy generation from biomass, demonstrating the potential of black liquor residue as an alternative energy source.

Synthesis/separation of 5-hydroxymethylfurfural converted from fructose promoted by H2O–CO2 biphasic system with solid catalysts: Experimental and kinetic modeling approaches

5-Hydroxymethylfurfural (5-HMF) is an industrially valuable compound typically produced by dehydration of fructose converted from cellulosic biomass. However, the formation of 5-HMF is accompanied by its conversion into by-products, such as levulinic and formic acids, decreasing its yield. Therefore, biphasic processes are attracting much attention allowing the efficient simultaneous synthesis and separation of 5-HMF. In this study, a biphasic system using water with high-pressure carbon dioxide, the most environmentally friendly solvents, combining with a solid catalyst is used to achieve 5-HMF synthesis and separation at lower temperatures and shorter duration than previous studies. The main experiment was operated by a semibatch reactor with a continuous flow of CO2. Additionally, three experiments, i) measurement of the partition coefficient of 5-HMF between the two phases, ii) 5-HMF synthesis in H2O–CO2 biphasic batch system, and iii) 5-HMF extraction in H2O–CO2 biphasic semibatch system, were conducted to complement the main experiment. All results from complement experiments were used as parameters in the kinetic modeling. As the result, the maximum yield of 5-HMF (36.6 %) was achieved at a temperature of 403 K, a pressure of 25 MPa, and a reaction time of 180 min. Finally, the optimal conditions for the semibatch experiment for 5-HMF synthesis and separation could be estimated from the kinetic models with parameters determined in this work. The obtained results suggested that this yield can be further improved by increasing the reaction time, which should be confirmed by further experimental studies.

Effects of binders and die geometry on quality of densified rice bran using a screw-type laboratory scale pelleting machine

Cellulosic biomass could solve environmental challenges posed by utilization of fossil fuels. Biomass needs to be processed through densification to ease handling, transport and storage. This study considered the densification of rice bran into pellets using a screw-type pelleting machine. Investigations determined the effect of the starch binder quantity, compression ratio and die configuration on the properties of the pellets. The bulk density, pellet length, pellet density, compressive strength, durability and higher heating value of the rice bran pellets ranged between 254 and 367 kg/m3, 16.85 to 39.90 mm, 283.42 and 793.87 kg/m3, 1.68 and 8.31 N/mm2, 89.06 and 98.32 %; and 16.54 to 17.30 MJ/kg. The binder quantity significantly determined the length of the rice bran pellets. The bulk density was significantly influenced by binder quantity, die type and compression ratio. The higher heating value of the rice bran pellets was significantly influenced by the compression ratio and binder quantity. The study established that interactions of the variables considered had influence on the pellet length and bulk density of pellets.

Experimental study combined with RSM process optimization for removal of the (Safranin O) cationic dye in the aqueous solution using a hydrogel prepared based on cellulosic biomass: an effective and ecological approach

Experimental study combined with RSM process optimization for removal of the (Safranin O) cationic dye in the aqueous solution using a hydrogel prepared based on cellulosic biomass: an effective and ecological approach

Study on Preparation of Regenerated Cellulose Fiber from Biomass Based on Mixed Solvents.

In this study, Arundo donax Linnaeus was utilized as the biomass and a TH/DS (Tetra-n-butylammonium hydroxide/Dimethyl sulfoxide, C16H37NO/C2H6OS) system was employed to dissolve biomass cellulose. The optimal process for the preparation of Arundo donax L. biomass regenerated cellulose fiber was determined through process optimization. The physical properties and antimicrobial performance of the resulting products were analyzed. The results demonstrated that the physical indicators of biomass regenerated cellulose fiber, prepared from Arundo donax L. cellulose, met the requirements of the standard for Viscose Filament (Dry breaking strength ≥ 1.65 CN/dtex, Elongation at dry breaking 15.5-26.0%, and Dry elongation CV value ≤ 10.0%). Additionally, excellent antimicrobial properties were exhibited by the biomass regenerated cellulose fiber developed in this study, with antibacterial rates against Staphylococcus aureus and other three strain indexes meeting the Viscose Filament standards. Furthermore, high antiviral activity of 99.99% against H1N1 and H3N2 strains of influenza A virus was observed in the experimental samples, indicating a remarkable antiviral effect. Valuable references for the comprehensive utilization of Arundo donax L. biomass resources are provided by this research.

Influence of Cellulose and Lignin-Rich Biomass on Catalyst Performance: A Study with Walnut Shell and Corn Stover Gasification

Influence of Cellulose and Lignin-Rich Biomass on Catalyst Performance: A Study with Walnut Shell and Corn Stover Gasification

Constructing green, smart wettability-switchable and antibacterial Caragana cellulose filter by cooperative regulation of texture and surface for on-demand purification of emulsions

The efficient on-demand purification of complex oily emulsions containing multiple pollutants and bacteria by a green filter has always been desired, but also extremely challenging. The abundant and renewable natural cellulose biomass is the best choice for developing green separation materials, but the limited separation capability of raw biomass restricts its application. To resolve this problem, this paper proposed a synergetic texture-surface regulation strategy to improve the liquid-locking ability and amphiphilicity of Caragana cellulose fiber, and thus prepared an intelligent multifunctional filter with an arbitrarily switchable underoil superhydrophobic (WCA = 152.1°) and underwater superoleophobic (OCA = 151.6°) surface. The surface wettability of the filter can be switched randomly by changing the wetting liquid, thus achieving efficient separation of different types of emulsions. As expected, both simulated different oils-in-water and different actual waters-in-oil emulsions, and all types of practical industrial oily complex emulsion sewage were purified by the filter at a high level (separation efficiency: 99.9 %). In addition, the filter can simultaneously remove 99.1 % of the Methylene blue and 98.6 % of the Oil red O in emulsions and has excellent antibacterial activity against E. coli (efficiency: 99.87 %) and S. aureus (efficiency: 99.43 %). The spent filter can be regenerated as an adsorbent capable of dynamically adsorbing 99.9 % of Orange II and 99.8 % of Methylene blue. This study provides a new method to modulate natural renewable cellulose and thus produce a multifunctional separation material for on-demand purification of complex and stubborn emulsions.

Mechanism of initial activation of carbon–oxygen bonds for deoxidation of acetic acid

Producing valuable fuel-ranged hydrocarbons from cellulosic biomass is an effective approach to addressing potential energy issues. However, the cellulose-derived bio-oil may contain carboxylic acid components that contribute to the unfavorable acidic and corrosive nature, which necessitates upgrading through the hydrodeoxygenation (HDO) treatment using heterogeneous catalysis. Here, we performed reactive force-field molecular dynamics (ReaxFF-MD) to study the hydrogenolysis, dehydrogenation and decarboxylation mechanisms of the acetic acid model compound on the Ni-based catalyst. Comparative analysis of the activation of C–O and C=O bonds was presented, followed by an examination of the associative desorption of H2 and the dynamic evolution of main intermediates. Furthermore, the conversion pathways of key intermediates were determined and four main pathways for ethanol generation and the temperature-response mechanism for coking formation were proposed. This mechanism diagram explains the strong effects of a hydrogen environment in biomass-related reactions, highlighting the dynamic mechanism of selective hydrogenation and deoxygenation of carbon–oxygen bonds.

Impact of Synergy Partner Cel7B on Cel7A Binding Rates: Insights from Single-Molecule Data.

Enzymatic degradation of cellulosic biomass is a well-established route for the sustainable production of biofuels, chemicals, and materials. A strategy employed by nature and industry to achieve an efficient degradation of cellulose is that cellobiohydrolases (or exocellulases), such as Cel7A, work synergistically with endoglucanases, such as Cel7B, to achieve the complete degradation of cellulose. However, a complete mechanistic understanding of this exo-endo synergy is still lacking. Here, we used single-molecule fluorescence microscopy to quantify the binding kinetics of Cel7A on cellulose when it is acting alone on the cellulose fibrils and in the presence of its synergy partner, the endoglucanase Cel7B. To this end, we used a fluorescently tagged Cel7A and studied its binding in the presence of the unlabeled Cel7B. This provided the single-molecule data necessary for the estimation of the rate constants of association kON and dissociation kOFF of Cel7A for the substrate. We show that the presence of Cel7B does not impact the dissociation rate constant, kOFF. But, the association rate of Cel7A decreases by a factor of 2 when Cel7B is present at a molar proportion of 10:1. This ratio has previously been shown to lead to synergy. This decrease in association rate is observed in a wide range of total enzyme concentrations, from sub nM to μM concentrations. This decrease in kON is consistent with the formation of cellulase clusters recently observed by others using atomic force microscopy.

Engineering of Ogataea polymorpha strains with ability for high-temperature alcoholic fermentation of cellobiose.

Successful conversion of cellulosic biomass into biofuels requires organisms capable of efficiently utilizing xylose as well as cellodextrins and glucose. Ogataea (Hansenula) polymorpha is the natural xylose-metabolizing organism and is one of the most thermotolerant yeasts known, with a maximum growth temperature above 50°C. Cellobiose-fermenting strains, derivatives of an improved ethanol producer from xylose O.polymorpha BEP/cat8∆, were constructed in this work by the introduction of heterologous genes encoding cellodextrin transporters (CDTs) and intracellular enzymes (β-glucosidase or cellobiose phosphorylase) that hydrolyze cellobiose. For this purpose, the genes gh1-1 of β-glucosidase, CDT-1m and CDT-2m of cellodextrin transporters from Neurospora crassa and the CBP gene coding for cellobiose phosphorylase from Saccharophagus degradans, were successfully expressed in O. polymorpha. Through metabolic engineering and mutagenesis, strains BEP/cat8∆/gh1-1/CDT-1m and BEP/cat8∆/CBP-1/CDT-2mAM were developed, showing improved parameters for high-temperature alcoholic fermentation of cellobiose. The study highlights the need for further optimization to enhance ethanol yields and elucidate cellobiose metabolism intricacies in O. polymorpha yeast. This is the first report of the successful development of stable methylotrophic thermotolerant strains of O.polymorpha capable of coutilizing cellobiose, glucose, and xylose under high-temperature alcoholic fermentation conditions at 45°C.

Hydrolysis of ionic liquid–treated substrate with an Iocasia fonsfrigidae strain SP3-1 endoglucanase

Recently, we reported the discovery of a novel endoglucanase of the glycoside hydrolase family 12 (GH12), designated IfCelS12A, from the haloalkaliphilic anaerobic bacterium Iocasia fonsfrigidae strain SP3-1, which was isolated from a hypersaline pond in the Samut Sakhon province of Thailand (ca. 2017). IfCelS12A exhibits high substrate specificity on carboxymethyl cellulose and amorphous cellulose but low substrate specificity on b-1,3;1,4-glucan. Unlike some endoglucanases of the GH12 family, IfCelS12A does not exhibit hydrolytic activity on crystalline cellulose (i.e., Avicel™). High-Pressure Liquid Chromatography (HPLC) and Thin Layer Chromatography (TLC) analyses of products resulting from IfCelS12-mediated hydrolysis indicate mode of action for this enzyme. Notably, IfCelS12A preferentially hydrolyzes cellotetraoses, cellopentaoses, and cellohexaoses with negligible activity on cellobiose or cellotriose. Kinetic analysis with cellopentaose and barely b-d-glucan as cellulosic substrates were conducted. On cellopentaose, IfCelS12A demonstrates a 16-fold increase in activity (KM = 0.27 mM; kcat = 0.36 s−1; kcat/KM = 1.34 mM−1 s−1) compared to the enzymatic hydrolysis of barley b-d-glucan (KM: 0.04 mM, kcat: 0.51 s−1, kcat/KM = 0.08 mM−1 s−1). Moreover, IfCelS12A enzymatic efficacy is stable in hypersaline sodium chlorids (NaCl) solutions (up to 10% NaCl). Specifically, IfCel12A retains notable activity after 24 h at 2M NaCl (10% saline solution). IfCelS12A used as a cocktail component with other cellulolytic enzymes and in conjunction with mobile sequestration platform technology offers additional options for deconstruction of ionic liquid–pretreated cellulosic feedstock.Key points• IfCelS12A from an anaerobic alkaliphile Iocasia fronsfrigidae shows salt tolerance• IfCelS12A in cocktails with other enzymes efficiently degrades cellulosic biomass• IfCelS12A used with mobile enzyme sequestration platforms enhances hydrolysis

Isolation and characterization of microalgae strains able to grow on complex biomass hydrolysate for industrial application

Producing algae biomass economically remains one of the chief bottlenecks for commercializing algae products. The aim of this work is to identify new strains of algae that can grow on cost effective media derived from cellulosic waste streams, characterize the potential of these strains to produce compounds of high industrial value, and identify those strains capable of facile genetic transformation. Here we report, out of 45 strains initially isolated, three were selected based on their ability to efficiently grow on organic waste material (corn stover hydrolysate) as a carbon source; Chlorococcum sp., Desmodesmus sp., and Chlamydomonas debaryana. Untargeted metabolomics was performed on each strain, identifying several metabolites of high relative abundances that are of commercial interest, such as lactic acid, butane-2,3-diol, amino acids, tartaric acid, triacylglycerols, and lipid species containing different, mono- and polyunsaturated fatty acids, depending on the strain and growth condition. The strains also produced carbohydrates of industrial relevance. Chlorococcum sp. was found to be genetically transformable using standard simple transformation protocols. These results suggest that with further development, these strains could open the door to economic production of high value commercial compounds utilizing waste streams from cellulosic biomass.