Felipe Antonio Fernandes Antunes

Bioconversion of sugarcane biomass into ethanol: an overview about composition, pretreatment methods, detoxification of hydrolysates, enzymatic saccharification, and ethanol fermentation.

Depleted supplies of fossil fuel, regular price hikes of gasoline, and environmental damage have necessitated the search for economic and eco-benign alternative of gasoline. Ethanol is produced from food/feed-based substrates (grains, sugars, and molasses), and its application as an energy source does not seem fit for long term due to the increasing fuel, food, feed, and other needs. These concerns have enforced to explore the alternative means of cost competitive and sustainable supply of biofuel. Sugarcane residues, sugarcane bagasse (SB), and straw (SS) could be the ideal feedstock for the second-generation (2G) ethanol production. These raw materials are rich in carbohydrates and renewable and do not compete with food/feed demands. However, the efficient bioconversion of SB/SS (efficient pretreatment technology, depolymerization of cellulose, and fermentation of released sugars) remains challenging to commercialize the cellulosic ethanol. Among the technological challenges, robust pretreatment and development of efficient bioconversion process (implicating suitable ethanol producing strains converting pentose and hexose sugars) have a key role to play. This paper aims to review the compositional profile of SB and SS, pretreatment methods of cane biomass, detoxification methods for the purification of hydrolysates, enzymatic hydrolysis, and the fermentation of released sugars for ethanol production.

Ultra-structural mapping of sugarcane bagasse after oxalic acid fiber expansion (OAFEX) and ethanol production by Candida shehatae and Saccharomyces cerevisiae

BackgroundDiminishing supplies of fossil fuels and oil spills are rousing to explore the alternative sources of energy that can be produced from non-food/feed-based substrates. Due to its abundance, sugarcane bagasse (SB) could be a model substrate for the second-generation biofuel cellulosic ethanol. However, the efficient bioconversion of SB remains a challenge for the commercial production of cellulosic ethanol. We hypothesized that oxalic-acid-mediated thermochemical pretreatment (OAFEX) would overcome the native recalcitrance of SB by enhancing the cellulase amenability toward the embedded cellulosic microfibrils.ResultsOAFEX treatment revealed the solubilization of hemicellulose releasing sugars (12.56 g/l xylose and 1.85 g/l glucose), leaving cellulignin in an accessible form for enzymatic hydrolysis. The highest hydrolytic efficiency (66.51%) of cellulignin was achieved by enzymatic hydrolysis (Celluclast 1.5 L and Novozym 188). The ultrastructure characterization of SB using scanning electron microscopy (SEM), atomic force microscopy (AFM), Raman spectroscopy, Fourier transform–near infrared spectroscopy (FT-NIR), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) revealed structural differences before and after OAFEX treatment with enzymatic hydrolysis. Furthermore, fermentation mediated by C. shehatae UFMG HM52.2 and S. cerevisiae 174 showed fuel ethanol production from detoxified acid (3.2 g/l, yield 0.353 g/g; 0.52 g/l, yield, 0.246 g/g) and enzymatic hydrolysates (4.83 g/l, yield, 0.28 g/g; 6.6 g/l, yield 0.46 g/g).ConclusionsOAFEX treatment revealed marked hemicellulose degradation, improving the cellulases’ ability to access the cellulignin and release fermentable sugars from the pretreated substrate. The ultrastructure of SB after OAFEX and enzymatic hydrolysis of cellulignin established thorough insights at the molecular level.

Dilute Acid Hydrolysis of Agro-Residues for the Depolymerization of Hemicellulose: State-of-the-Art

Geo-political, long-term economic and sustainable concerns are promoting researchers and entrepreneurs to harness the potential of lignocellulosic feedstock (LCF) into industrially significant products. Agro-residues (sugarcane bagasse, wheat straw, rice straw, corn stover, etc.) constitute the principal fraction of LCF and are available in large amounts globally. The judicious exploration of agro-residues into important products such as d-xylitol, an artificial sweetener, may provide a strong platform for its sustainable supply to the medical and non-medical applications-based sectors. Pretreatment of agro-residues by dilute acid hydrolysis is an inevitable process for the depolymerisation of hemicellulosic fraction into xylose and other sugars. Dilute acid catalyses hemicellulose fractionation at high temperature within short reaction times. Significant developments have been made in the past towards the chemical hydrolysis of agro-residues, particularly for the hemicellulose breakdown. Critical parameters such as acid load, temperature, residence time and solid-to-liquid ratio play pivotal roles in the kinetics of dilute acid hydrolysis of agro-residues. Furthermore, reactor configurations such as counter-current, plug-flow, percolation and shrinking-bed have been designed in order to maximize the sugars recovery with minimum inhibitors generation. This chapter reviews the process parameters, kinetics, methods and reactor engineering for the dilute acid catalysed processes employed for agro-residues.

Bioethanol Production from Sugarcane Bagasse by a Novel Brazilian Pentose Fermenting Yeast Scheffersomyces shehatae UFMG-HM 52.2: Evaluation of Fermentation Medium

Bioconversion of hemicellulosic sugars into second generation (2G) ethanol plays a pivotal role in the overall success of biorefineries. In this study, ethanol production performance of a novel xylose-fermenting yeast, Scheffersomyces shehatae UFMG-HM 52.2, was evaluated under batch fermentation conditions using sugarcane bagasse (SB) hemicellulosic hydrolysate as carbon source. Dilute acid hydrolysis of SB was performed to obtain sugarcane bagasse hemicellulosic hydrolysate (SBHH). It was concentrated, detoxified, and supplemented with nutrients in different formulations to prepare the fermentation medium to the yeast evaluation performance. S. shehatae UFMG-HM 52.2 (isolated from Brazilian Atlantic rain forest ecosystem) was used in fermentations carried out in Erlenmeyer flasks maintained in a rotator shaker at 30°C and 200 rpm for 72 h. The use of a fermentation medium composed of SBHH supplemented with 5 g/L ammonium sulfate, 3 g/L yeast extract, and 3 g/L malt extract resulted in 0.38 g/g of ethanol yield and 0.19 g L.h of volumetric productivity after 48 h of incubation time.

Current applications and different approaches for microbial L-asparaginase production

l-asparaginase (EC 3.5.1.1) is an enzyme that catalysis mainly the asparagine hydrolysis in l-aspartic acid and ammonium. This enzyme is presented in different organisms, such as microorganisms, vegetal, and some animals, including certain rodents serum, but not unveiled in humans. It can be used as important chemotherapeutic agent for the treatment of a variety of lymphoproliferative disorders and lymphomas (particularly acute lymphoblastic leukemia (ALL) and Hodgkins lymphoma), and has been a pivotal agent in chemotherapy protocols from around 30 years. Also, other important application is in food industry, by using the properties of this enzyme to reduce acrylamide levels in commercial fried foods, maintaining their characteristics (color, flavor, texture, security, etc.) Actually, l-asparaginase catalyzes the hydrolysis of l-asparagine, not allowing the reaction of reducing sugars with this aminoacid for the generation of acrylamide. Currently, production of l-asparaginase is mainly based in biotechnological production by using some bacteria. However, industrial production also needs research work aiming to obtain better production yields, as well as novel process by applying different microorganisms to increase the range of applications of the produced enzyme. Within this context, this mini-review presents l-asparaginase applications, production by different microorganisms and some limitations, current investigations, as well as some challenges to be achieved for profitable industrial production.

Hydrodynamic cavitation as an efficient pretreatment method for lignocellulosic biomass: A parametric study

Hydrodynamic cavitation (HC), which is a highly destructive force, was employed for pretreatment of sugarcane bagasse (SCB). The efficacy of HC was studied using response surface methodology (RSM) with determining parameters varied: inlet pressure of 1-3bar, temperature of 40-70°C, and alkaline concentration of 0.1-0.3M. At the best condition (3bar, 70°C and 0.3M NaOH), 93.05% and 94.45% of hydrolysis yield of cellulose and hemicellulose, respectively, were obtained within 30min of pretreatment time. Also, pretreatment time higher than 10min had little to do regarding to SCB composition changes using different orifice plates (16 and 27 holes, with corresponding cavitation number of 0.017 and 0.048, respectively), with higher hydrolysis yield observed at 20min of process. Therefore, HC-based approach could lead to a high yield of hydrolysis, as long as a treatment condition was right; it could be so at mild conditions and at short running time.

Hemicellulosic ethanol production by immobilized cells of Scheffersomyces stipitis: Effect of cell concentration and stirring

Bioconversion of hemicellulosic hydrolysate into ethanol plays a pivotal role in the overall success of biorefineries. For the efficient fermentative conversion of hemicellulosic hydrolysates into ethanol, the use of immobilized cells system could provide the enhanced ethanol productivities with significant time savings. Here, we investigated the effect of 2 important factors (e.g., cell concentration and stirring) on ethanol production from sugarcane bagasse hydrolysate using the yeast Scheffersomyces stipitis immobilized in calcium alginate matrix. A 22 full factorial design of experiment was performed considering the process variables- immobilized cell concentration (3.0, 6.5 and 10.0 g/L) and stirring (100, 200 and 300 rpm). Statistical analysis showed that stirring has the major influence on ethanol production. Maximum ethanol production (8.90 g/l) with ethanol yield (Yp/s) of 0.33 g/g and ethanol productivity (Qp) of 0.185 g/l/h was obtained under the optimized process conditions (10.0 g/L of cells and 100 rpm).

Ethanol production in a simultaneous saccharification and fermentation process with interconnected reactors employing hydrodynamic cavitation-pretreated sugarcane bagasse as raw material

In this study, sugarcane bagasse (SCB) pretreated with alkali assisted hydrodynamic cavitation (HC) was investigated for simultaneous saccharification and fermentation (SSF) process for bioethanol production in interconnected column reactors using immobilized Scheffersomyces stipitis NRRL-Y7124. Initially, HC was employed for the evaluation of the reagent used in alkaline pretreatment. Alkalis (NaOH, KOH, Na2CO3, Ca(OH)2) and NaOH recycled black liquor (successive batches) were used and their pretreatment effectiveness was assessed considering the solid composition and its enzymatic digestibility. In SSF process using NaOH-HC pretreatment SCB, 62.33% of total carbohydrate fractions were hydrolyzed and 17.26g/L of ethanol production (0.48g of ethanol/g of glucose and xylose consumed) was achieved. This proposed scheme of HC-assisted NaOH pretreatment together with our interconnected column reactors showed to be an interesting new approach for biorefineries.

Pretreatment of Sugarcane Bagasse and Leaves: Unlocking the Treasury of “Green Currency”

Sugarcane residues (bagasse and leaves/trash) are the principal feedstock in Asia, South America, Africa, and other parts of the world. The judicious application of this feedstock into value-added products such as fuel ethanol, xylitol, organic acids, industrial enzymes, etc. may provide a strong economic platform along with clean and safe environment. Pretreatment is an inevitable process to harness the carbohydrate fraction of sugarcane bagasse and leaves into readily available sugars by cellulase-mediated process for the production of house-hold commodities. Several methods (physical, physico-chemical, chemical, and biological) have been adopted for the pretreatment of sugarcane residues. Pretreatment methods with pros and cons are employed either to depolymerize hemicellulosic fraction or lignin degradation to make cellulose more amenable for improved cellulolytic enzymes action. The choice of pretreatment methods depends upon its precise mechanistic action on lignin or hemicelluloses with fewer inhibitory products, minimal sugar loss by increasing the cellulosic surface area for subsequent enzymatic action to obtain desired sugars recovery. Furthermore, economics and environmental impacts are two important considerations for the selection of pretreatment method. This chapter aims to explore a better understanding of multiple pretreatment methodologies applied to the sugarcane residues along with economics and environmental impacts.

Biosurfactant production by Aureobasidium pullulans in stirred tank bioreactor: New approach to understand the influence of important variables in the process

Surfactants are amphiphilic molecules with large industrial applications produced currently by chemical routes mainly derived from oil industry. However, biotechnological process, aimed to develop new sustainable process configurations by using favorable microorganisms, already requires investigations in more details. Thus, we present a novel approach for biosurfactant production using the promising yeast Aureobasidium pullulans LB 83, in stirred tank reactor. A central composite face-centered design was carried out to evaluate the effect of the aeration rate (0.1-1.1min-1) and sucrose concentration (20-80g.L-1) in the biosurfactant maximum tensoactivity and productivity. Statistical analysis showed that the use of variables at high levels enhanced tensoactivity, showing 8.05cm in the oil spread test and productivity of 0.0838cm.h-1. Also, unprecedented investigation of aeration rate and sucrose concentration relevance in biosurfactant production by A. pullulans in stirred tank reactor was detailed, demonstrating the importance to establish adequate conditions in bioreactors, aimed to scale-up process.