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The Amazon rainforest has a rich biodiversity, and studies of Basidiomycete fungi that have biomolecules of biotechnological interest are relevant. The use of lignocellulosic biomass in biotechnological processes proposes an alternative use, and also adds value to the material when employed in the bioconversion of agro-industrial waste. In this context, this study evaluate the production of lignocellulolytic enzymes (carboxymethylcellulases (CMCase), xylanase, pectinase, laccase) as well as phenolic compounds and proteases by solid-state fermentation (SSF) using the fungus Lentinus strigosus isolated from Amazon. The guarana (Paullinia cupana) residue was characterized using scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). SSF was carried out with 60% humidification of the residue, at 30 °C, for 10 days. The lignocellulosic biomass presented fragmented structures with irregular shapes and porosities, and was mainly constituted by cellulose (19.16%), hemicellulose (32.83%), and lignin (6.06%). During the SSF, significant values of CMCase (0.84 U/g) on the 8th day, xylanase (1.00 U/g) on the 7th day, pectinase (2.19 U/g) on the 6th day, laccase (176.23 U/mL) on the 5th day, phenolic compounds (10.27 μg/mL) on the 1st day, soluble proteins (0.08 mg/mL) on the 5th day, and protease (8.30 U/mL) on the 6th day were observed. In general, the agro-industrial residue used provided promising results as a viable alternative for use as a substrate in biotechnological processes.

The bioalcohol production (ethanol and butanol) from lignocellulosic biomass has some techno-economic limitations nowadays. Detoxification technologies can efficiently remove fermentation inhibitors (such as furan aldehydes, aliphatic acids, and phenolic compounds) to increase the bioalcohol titers. However, the addition of more steps before fermentation increases the complexity of the production scheme and reduces the profit margin. Process intensification concepts have been disseminated as a critical factor for the success of biotechnological processes, including bioalcohol production. Thus, this review discusses the main inhibitors from lignocellulosic biomass and the in-situ technologies used to minimize the yeast inhibition in hydrolysates and slurries. In-situ detoxification strategies involve different fundamentals, but they have the unanimity of allowing fermentation to be carried out concurrently. They are usually based on the establishment of a microbial consortium as well as the addition of reducing agents, polymers, solvents and adsorbents. Also, in-situ product recovery methods have been developed to minimize energy consumption in downstream processing.

Filmogenic coatings can be used as a post-harvest strategy to extend shelf life and ensure improvements in fruit quality and safety given their perishable nature. Due to their edibility, the composition of the coverings is a determining factor for their application, and for this reason, it is interesting and desirable that the constituents come from natural sources. The objective of the present study was to develop and characterize a pectin-based film extracted from cajá peel (Spondias mombin) and verify its efficiency as an edible coating in postharvest acerolas (Malpighia emarginata). The film was characterized in terms of mechanical properties by Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM); as well as in terms of chemical properties using the analysis of structural bonds by Fourier Transformed Infrared Spectroscopy (FTIR) and X-Ray Diffraction (XRD). In addition, the antimicrobial activity against two bacteria was evaluated. The analyzed film was presented as a dense membrane, with the presence of pores, fissures, and a very rough surface. The degree of esterification of pectin extract from cajá peel was 44%, and for this reason it was classified as low methocxyl (LM) pectin. The filmogenic solution presented antimicrobial activity against the bacteria Pseudomonas aeruginosa and Staphylococcus aureus. The applicability of the edible coating was tested on green acerolas, monitoring their maturation stage through analyzes such as weight loss, titratable acidity, and total soluble solids. At the end of the 7 days of storage, acerolas with the application of the coating showed 8.97% weight loss while acerolas without coating showed 9.89%; the percentage of total soluble solids was 7.68% higher for acerolas with the coating, as well as ascorbic acid content was higher for the protected fruits, indicating that the coverage favored the delay in the maturation of acerolas.

Trichoderma reesei is a fungus that has been widely explored for its potential as cellulolytic enzyme producer and has diverse industrial applications. However, obtaining the enzymes is still considered a costly and, sometimes, inefficient process. This study aimed to produce endoglucanases by cultivating T. reesei (CCT-2768) in solid state fermentation, using cashew apple bagasse (CAB), sugarcane bagasse (SCB) and green coconut fiber (GCF) residues as substrates. The influence of moisture and pH on enzyme production was evaluated using a factorial design. T. reesei showed viability for producing endoglucanases in all of the three lignocellulosic residues tested, with maximum activity (2.29 ± 0.01 U/g) observed when cultivated in the SCB substrate and using moisture of 60% and pH 5.5. Thus, use of lignocellulosic residues proves to be a viable alternative for producing endoglucanases by cultivation of Trichoderma reesei, which contributes to the recycling of waste and the reduction of environmental impacts.

The addition of poly(ethylene glycol) (PEG) in the pretreatment and enzymatic hydrolysis steps has become attractive to increase second-generation ethanol titers. Recent studies have shown the benefits of using high-concentration PEG on the yeast vitality and enzymatic digestibility of untreated biomass. Thus, this study investigated the ideal concentration of PEG in the second-generation ethanol production and the best step to add it. In the acid pretreatment, PEG 4000 favored the delignification (59.2%) and reduced the non-productive adsorption of cellulases onto corn cob, but its use was not significant on the sodium carbonate–based pretreatment. Changes in PEG dosage and PEG molecular weight strongly affected the performance of enzymatic hydrolysis and simultaneous saccharification and fermentation (SSF). PEG 4000 (150 g/L) increased the production of reducing sugars by up to 30.1% and the ethanol production via SSF by up to 44.7% from untreated corn cob. In turn, PEG slightly affected the enzymatic digestibility and fermentability of the pretreated corn cob. One-pot ethanol production from corn cob was performed using PEG as an additive. The addition of 150 g/L PEG 1500 and PEG 4000 increased the ethanol yield values (> 52%) in the one-pot processes; however, the ethanol concentration was lower than in the experiments with the pretreated corn cob. SSF experiments with high solid loadings were tested using PEG 1500. Under these conditions, PEG 1500 slightly increased ethanol production, from 53.4 to 55.7 g/L ethanol (equivalent to 92.8% ethanol yield) using 20% (w/v) acid-pretreated corn cob.

Hyaluronic acid (HA) is a biopolymer with applications in different areas such as medicine and cosmetics. HA is currently either isolated from animal sources or produced by microbial fermentation. Animal HA presents some disadvantages such as high cost and risk of viral cross-species or another infectious agent. In the present study, we evaluated the physicochemical characteristics and in vitro antioxidant capacity of HA produced by Streptococcus zooepidemicus CCT 7546. In addition, commercial sodium hyaluronate (SH) from an animal source was used as control. The microbial HA yield after purification was 69.8 mg/L. According to Fourier transform infrared spectroscopy, it was seen that bacterial and animal HA spectra are overlapped. The thermogravimetric analysis revealed that microbial HA was more stable than its equivalent from the animal source. However, scanning electron microscopy indicates that the purification method used in the animal product was more effective. Microbial HA showed activity in total antioxidant capacity (14.02 ± 0.38%), reducing power (18.18 ± 6.43%), DPPH radical-scavenging (5.57 ± 0.23 kmol TE/g), and hydroxyl radical-scavenging (28.39 ± 2.40%) tests. Therefore, in vitro antioxidant tests demonstrated that the antioxidant action mechanism occurs through scavenging reactive oxygen species (ROS) and donating electrons/hydrogen atoms.

Lignocellulosic biomass is the most abundant biological resource on the planet and has been extensively researched to produce cellulosic ethanol. However, there is a consensus that the presence of lignin hinders the biomass conversion. Lignin is often considered a villain in cellulosic ethanol production studies due to its adverse effects on cellulases and yeasts. Despite this, recent studies indicate that lignins can be transformed into useful inputs to produce cellulosic ethanol. These approaches aim to establish closed-loop biorefineries to improve economic metrics and reduce the environmental impact due to the substitution of products based on fossil sources. The present review addresses the successful cases in transforming lignin into chemicals and materials to increase cellulosic ethanol titers. A contextualization was first carried out, considering aspects of biomass characteristics and lignin valorization. The impact of lignin-based chemicals and materials in the pretreatment, detoxification, and enzymatic hydrolysis steps was discussed in detail. Economic aspects and future perspectives were also included in this review. These reports open a new point of view on lignin valorization and its integration with the cellulosic ethanol production chain.

The present study focuses on elucidating the effects of ethylene oxide-based polymers on cellulosic ethanol production from green coconut fiber (GCF). Data on ethanol production, cell viability, adsorption of cellulases, and enzymatic digestibility were collected from experiments with high concentrations of PEG 4000 and poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol) 5800 (EOPO 5800). PEG 4000 and EOPO 5800 favored cell viability and ethanol production in simulated experiments with S. cerevisiae CAT-1 and Kluyveromyces marxianus. Enzymatic hydrolysis under 175 g L−1 PEG 4000 increased the sugar release using untreated GCF (702.0%), hydrothermally-pretreated GCF (63.2%), and acid-pretreated GCF (40.1%). However, there were no benefits to using SigmaCell cellulose as the substrate. EOPO 5800 at 100 g L−1 had positive effects on cellulolytic activity preservation and on the sugar release during enzymatic hydrolysis. Both polymers provided higher ethanol titers for simultaneous saccharification and fermentation (SSF) of pretreated GCF compared to polymer-free experiments. A high ethanol yield (89.8%) was reached by the SSF of untreated GCF using 175 g L−1 PEG 4000. This result implies that high polymer concentrations may enable the cellulosic ethanol scheme without the need for pretreatment.

Large volumes of produced water are generated as a byproduct in activities of oil and gas exploitation, which can be reused in agriculture after a treatment process. Activated sludge treatment has been successfully used to remove oil from wastewater, but systematic studies on the toxicity of this effluent using this treatment are scarce in the literature. In this study, it was investigated the performance of an activated sludge system in the treatment of a synthetic produced water under different initial conditions in terms of salinity and oil and grease concentration. Furthermore, it was evaluated this effluent phytotoxicity in the germination, and seedling and plant growths of sunflower and corn seeds using untreated and treated synthetic produced water. Results revealed the activated sludge effectiveness in oil and grease and salinity removal from produced water, viz. high removal efficiency of 99.01 ± 0.28 and 91.07 ± 0.39%., respectively. Untreated produced water showed considerable toxic effects on the germination (74.67 ± 2.31% and 82.67 ± 2.31 for sunflower and corn seeds, respectively) and growth stages of sunflower and corn seed plants. The germination percentage was approximately 100% for both types of seed. The seedling and plant growth of the two seeds irrigated with treated produced water had similar performance when used tap water. These results highlighted the potential reuse as an unconventional water resource for plant irrigation of the synthetic produced water treated by an activated sludge process, which technology has showed high removal performance of salinity and oil.

The present study aimed to evaluate the lactose hydrolysis conditions from “coalho” cheese whey using β-galactosidase (β-gal) produced by Kluyveromyces lactis immobilized with sodium alginate. Thre...

The objective of this work was to evaluate the effects of enzymatic hydrolysis on digestibility and morphological and structural properties of hydrothermally pre-treated (HPT) red rice starch. The pre-treatments were performed in autoclave and cooking for the modification of rice grains and native starch. In vitro starch digestibility was performed consecutively and semi-simultaneously using α-amylase and amyloglucosidase. A first-order mathematical model was used to adjust the hydrolysis kinetic data, which made it possible to calculate the surface area, hydrolysis index, and glycemic index of the starch. Scanning electron microscopy images (SEM), Fourier transform infrared (FTIR) spectra and X-ray diffraction (XRD) were also performed to investigate the characteristics of the post-hydrolysis starch samples. The autoclaved starch HSS-A3, which was subjected to 121 °C/1.08 bar for 10 min, showed the highest in vitro digestibility values (80.08 %). Both starch samples showed increase of particle size and enzymatic digestibility after HPT. FTIR spectra of the starch samples showed that there was no appearance of new functional groups. However, XRD evidenced that HPT changed the intensity of the peaks and the type of crystallinity was changed for autoclaved starch (A3) from type A to Vh, with crystallinity ranging from 21.71 % to 26.42 %. The semi-simultaneous approach showed more advantages due to the highest in vitro digestibility as well as reducing the processing time and use of reagents.

The objective of this study was to evaluate the modification of red rice starch by a combination of hydrothermal pretreatments and α-amylase hydrolysis. In vitro digestibility and the morphological, structural, functional, thermal, textural and rheological properties of red rice starch were evaluated. The starch submitted to autoclave (A3) obtained the highest hydrolysis yield (37.66 %) after 300 min. The morphological analysis showed that for the native starch, the granules presented a polyhedral shape and increased in diameter (2.36-394.12 μm) due to hydrothermal pre-treatments. α-Amylase (9 U mg

The present study proposes to obtain polygalacturonases, antioxidant compounds, and active chitosan films from a valorization scheme of mangaba residue. After extraction with ethanolic solution, the mangaba residue was used as a substrate for solid state fermentation (SSF) of Aspergillus niger IOC 4003, which obtained 76.32 U g−1 for polygalacturonase after 96 h cultivation. The previous step of solvent extraction increased the enzyme productivity and allowed the production of pH-tolerant polygalacturonase in SSF experiments. The addition of polygalacturosanase under 1.0 U mL−1 enabled the reduction of turbidity (34%) and viscosity (11%) of mangaba juice compared to enzyme-free condition. Solid residue from the SSF step was used in the extraction of alkaline lignin with sodium hydroxide, which allowed the recovery of 21.82% of the initial Klason lignin content. Supplementation of chitosan films with ethanolic extract and 0.1% (w v−1) alkaline lignin changed the values of antioxidant activity (increase of 86.5% and 45.4%) and permeability (reduction of 43.2% and 2.3%).

In this study, cellulose nanocrystals (CNC) were produced from untreated and pretreated sugarcane fibers through enzymatic hydrolysis using a crude cellulolytic extract produced by the fungus Aspergillus fumigatus CCT 7873 using semi-solid fermentation (SSF). The fibers were characterized in terms of cellulose, hemicellulose, and lignin contents and crystallinity index. The CNC generated during the enzymatic hydrolysis using a sugarcane fiber pretreated with 4% NaOH and an enzymatic loading of 7.5 FPU g−1 of bagasse were characterized by atomic force microscopy (AFM). The results showed that after the hydrolysis time of 48 h, spherical-like particles with a good size distribution and a mean diameter of 61 nm were obtained. In this study, we produced cellulose nanocrystals (CNC) from untreated and pretreated sugarcane fibers through enzymatic hydrolysis using a crude cellulolytic extract produced by the fungus Aspergillus fumigatus CCT 7873 using semi-solid fermentation (SSF). In fact, the main novelty of this manuscript is the fact that it accompanies the formation of CNC, by means of atomic force microscopy (AFM). It is noteworthy that there are few manuscripts with this approach in the literature. In addition, this was performed during the cultivation of a fermentation broth containing cellulases produced by an enzymatic extract of the fungus Aspergillus fumigatus CCT 7873 using SSF. Thus, CNC, a highly added value product, can be obtained from lignocellulosic residues, which is a renewable and low-cost source. The results showed that, during the cultivation, there is a variation in the size distribution of the CNC and that, for the hydrolysis time of 48 h, a good size distribution was obtained for the CNC with about 61 nm in diameter, presenting a spherical shape.

The present study evaluated the surfactin production by Bacillus subtilis UFPEDA 438 using sugarcane molasses as a substrate. The effects of the cultivation conditions (temperature, agitation and a...

2023-07 Lignocellulolytic enzymes have a great biotechnological potential. Their commercial exploitation has been considerably increased in recent years. However, its industrial application in the conversion of lignocellulosic biomass to biofuels depends on the development of more economical processes and technologies for the lignocellulolytic enzyme production. In this study, the improvement of cellulases and xylanases by Trichoderma reesei CCT-2768 production induced by pretreated carnauba straw with alkaline hydrogen peroxide (A-HP) was performed and its crude extract produced was applied in the enzymatic hydrolysis of sugarcane bagasse. Herein, three conditions were carried out consisting of changing solid loading, pretreatment time, and hydrogen peroxide concentration. The FPase, CMCase, xylanase, and β-glucosidase activities of produced crude extracts were measured. Pretreated carnauba straw had a higher production of total cellulase (2.4 U/g dry substrate) and xylanase (172 U/g dry substrate) enzymes. The low cellulases (8.0 U/g dry substrate) and high xylanase charges (544 U/g dry substrate) present in the produced extract and applied into pretreated sugarcane bagasse hydrolysis allowed a hydrolysis efficiency of 86.96%. The in situ lignocellulolytic enzyme production can represent a relevant advance in the future in overall cost reducing enzymatic hydrolysis process of lignocellulosic materials

The present study evaluated the co-production of β-galactosidase and ethanol by Kluyveromyces marxianus ATCC 36907 and Kluyveromyces lactis NRRL Y-8279 using as carbon source the lactose found on “...

Chitooligosaccharides (COS) have a great potential to be used by pharmaceutical industry due to their many biological activities. The use of enzymes to produce them is very advantageous, however it still faces many challenges, such as discovering new strains capable to produce enzymes that are able to generate bioactive oligosaccharides. In the present study a purification protein protocol was performed to purify chitosanases produced by Bacillus toyonensis CCT 7899 for further chitosan hydrolysis. The produced chitooligosaccharides were characterized by mass spectroscopy (MS) and their antiedematogenic effect was investigated through carrageenan-induced paw edema model. The animals were treated previously to inflammation by intragastric route with COS at 30, 300 and 600mg/kg. The purification protocol showed a good performance for the chitosanases purification using 0.20M NaCl solution to elute it, with a 9.54-fold purification factor. The treatment with COS promoted a decrease of paw edema at all evaluated times and the AUC04h, proving that COS produced showed activity in acute inflammation like commercial anti-inflammatory Dexamethasone (corticosteroid). Therefore, the strategy used to purification was successfully applied and it was possible to generate bioactive oligosaccharides with potential pharmacological use., The authors thank CAPES and Brazilian National Council for Research [Grant 401817/2016-4] for the financial supporting., info:eu-repo/semantics/publishedVersion

This paper establishes an efficient protocol for the immobilization of Thermomyces lanuginosus lipase (TLL) on a hydrophobic resin, Streamline phenyl. The biocatalyst produced by TLL immobilization on Streamline phenyl resin was named iTLL. In addition, strategies to improve stability and reusability of iTLL were performed using polyethylenimine (PEI) or/and glutaraldehyde (GA), producing iTLL-GA, iTLL-PEI, iTLL-PEI-GA biocatalysts. The immobilization yield was about 50%, using 1 mg/g of enzyme loading, and the immobilized enzyme activity was about 77 U/g, achieving about 100% of recovered activity. Desorption assays of the enzyme from the support using 0.6% cetyltrimethylammonium bromide (CTAB) and thermal and operational stability assays were performed. Although iTLL-PEI-GA lost about 50% of its initial activity after PEI and GA modifications, it was the most thermally and operationally stable (increases its stability about 66% if comparing with soluble enzyme at 65 ºC and maintenance 90% of its initial activity after 5 cycles of pNPB hydrolysis at 25 °C and pH 7.0). Furthermore, it showed almost no desorption of enzyme molecules with 24 h of CTAB incubation. Moreover, the streamline phenyl demonstrated a high TLL loading potential, with no diffusion limitations up to 14 mg/g. These characteristics allow future application of the iTLL-PEI-GA biocatalyst in fluidized bed reactors.

Toxoplasmosis, despite advances in science and technology, is a disease that requires attention since there is no vaccine capable of immunizing humans and animals against all isolated types of Toxoplasma gondii. Thus, the use of chimeric proteins, which can contain multiple antigens, is a very promising alternative for the process of obtaining a vaccine and diagnostic test for toxoplasmosis due to the great diversity of antigens presented by T. gondii. In this context, the present study evaluates batch culture strategies in the production of the multi-antigenic chimeric protein TgAGS/BsT from Toxoplasma gondii. Several exploratory cultures were initially carried out to observe the kinetic behavior of E. coli BL21 Star in five different medium compositions without the addition of IPTG (inducer). Cultures of E. coli B21 Star were carried out with 1.0 mM IPTG at different times of initiation of induction (0.5, 1, and 6 h) to evaluate the effects on cell growth, production of the protein of interest, culture pH, and acetic acid formation. The results showed that among the culture media evaluated, 2xTY and TB supplemented with glycerol had the best cell concentration values of 3.42 ± 0.05 g/L and 5.48 ± 0.05 g/L, respectively. In the assays induced by IPTG, a higher expression of TgAGS/BsT protein was observed, with induction beginning within 6 h of culture, with a maximum concentration of protein of interest of 1.82 ± 0.02 g/L for the 2xTY and 2.49 ± 0.03 g/L for the TB medium. In addition, later induction by IPTG provided greater stability of plasmid pET-TgAGS, remaining with values above 90% at the end of culture.

The improvement of biotechnological processes capable of transforming agro-industrial waste into products with high added value has stood out in the area of renewable energies, promoting positive impacts to the environment. Thus, the present work evaluated the influence of nickel on the conversion of fermentable sugars, present in the green coconut shell hydrolyzate (GCSH), into butanol and other products. Fermentation assays were performed at 37 °C, starting with 19.4 g.L-1 of sugars and 1.0 g.L-1 of inoculum (C. beijerinckii). The GCSH was supplemented with tryptone, yeast extract, ammonium acetate, minerals and phosphate buffer. Two conditions were tested: with and without addition of nickel. Concentrations of sugars (glucose and xylose), intermediate products (organic acids), acetone, butanol, and ethanol were determined by high performance liquid chromatography (HPLC). The results show that the butanol production was higher from GCSH without addition of nickel, reaching a concentration of 2.14 g.L-1 of butanol. Therefore, the presence of nickel in the hydrolyzate was not favorable in the production of butanol under the studied process conditions. The improvement of biotechnological processes capable of transforming agro-industrial waste into products with high added value has stood out in the area of renewable energies, promoting positive impacts to the environment. Thus, the present work evaluated the influence of nickel on the conversion of fermentable sugars, present in the green coconut shell hydrolyzate (GCSH), into butanol and other products. Fermentation assays were performed at 37 °C, starting with 19.4 g.L-1 of sugars and 1.0 g.L-1 of inoculum (C. beijerinckii). The GCSH was supplemented with tryptone, yeast extract, ammonium acetate, minerals and phosphate buffer. Two conditions were tested: with and without addition of nickel. Concentrations of sugars (glucose and xylose), intermediate products (organic acids), acetone, butanol, and ethanol were determined by high performance liquid chromatography (HPLC). The results show that the butanol production was higher from GCSH without addition of nickel, reaching a concentration of 2.14 g.L-1 of butanol. Therefore, the presence of nickel in the hydrolyzate was not favorable in the production of butanol under the studied process conditions.

Chitooligosaccharides (COS) have a great potential to be used by pharmaceutical industry due to their many biological activities. The use of enzymes to produce them is very advantageous, however it still faces many challenges, such as discovering new strains capable to produce enzymes that are able to generate bioactive oligosaccharides. In the present study a purification protein protocol was performed to purify chitosanases produced by

Hyaluronic acid (HA) is a biopolymer with applications in different areas such as medicine and cosmetics. HA is currently either isolated from animal sources or produced by microbial fermentation. Animal HA presents some disadvantages such as high cost and risk of viral cross-species or another infectious agent. In the present study, we evaluated the physicochemical characteristics and

In this study the use of the essential oil from a Lippiagracilis Schauer micro emulsion system as a biocide and antibiofouling agent upon biofilms as well as the effect of this micro emulsion on the corrosion rate of AISI 1020 carbon steel was investigated. The results showed that a microemulsion type Winsor-IV was efficient in preventing the biofouling formation after 96 hours of contact and inhibited the growth of the sulfate reducing, iron-oxidizing bacteria as well as the fungi forming the biofilms after the 16 days of contact time. The antimicrobial action was likely due to a formation of a protective film.

The present study aimed to evaluate the influence of induction conditions (IPTG concentration, temperature, and induction time) on the plasmid pQE-30 stability and 503 antigen expression of Leishmania i. chagasi in Escherichia coli M15. Batch cultures were performed at 37 °C and induced by the addition of different IPTG concentrations (0.01 to 1.5 mM). Subsequently, experiments were carried out at different temperatures (27 to 42 °C), evaluating the influence of induction time (0.5 to 6 h after the start of the culture). The results showed that IPTG toxicity caused a metabolic stress in the cells and, consequently, the microorganism growth reduced. The induction with IPTG may also be associated with the plasmid pQE-30 instability, due to metabolic burden imposed by the recombinant protein expression. The optimal conditions for 503 antigen expression of Leishmania i. chagasi in Escherichia coli M15 were an IPTG concentration of 1.0 mM, temperature of 37 °C, and induction time of 2 h. The maximum antigen concentration obtained was 0.119 ± 0.009 g/L, about seven times higher than the lowest concentration. Therefore, the results showed that 503 antigen can be produced in laboratory; however, it requires more studies to minimize the plasmid instability and improve to industrial scale.

In this study, we investigated four conventional pretreatments under pressurized conditions, as well as in situ detoxification using polyethylene glycol (PEG 400), for the simultaneous saccharification and fermentation (SSF) of green coconut fibers. The highest fermentable sugar production (22.05% g/g untreated fiber) was reached with pressurized dilute alkaline pretreatment. In the SSF, the addition of PEG favored ethanol production compared to that in experiments without the polymer, reaching a maximum value of 7.87 g/L at a liquor content of 50% v/v for the pressurized water pretreatment. After lignin recovery by acid precipitation, the liquors had interesting antioxidant properties, similar to those of fruit extracts. The proposed pressurized water pretreatment could be an advantageous choice because it can be performed at mild temperatures (70 °C) and high solid loadings (26.67% w/v) and achieves a high mass yield (72.84% w/w) without the addition of a chemical agent. Furthermore, the liquor can be used in the SSF, reducing the water consumption and number of steps in the cellulosic ethanol process.

The 503-antigen, the elongation factor (1-γ) found in the amastigote phase of Leishmania infantum chagasi, could integrate either kits of specific diagnostic assays or vaccines for visceral leishmaniasis. However, protein purification expressed in Escherichia coli is driven by several steps, namely, clarification and multisteps chromatography. Additionally, the removal of lipopolysaccharide (LPS/endotoxin) which triggers an immune response in the human body is needed since LPS interacts with the proteins impairing the chromatography steps. Aqueous two-phase systems (ATPSs) based on polyethylene glycol (PEG) 1500 and salt (K2HPO4 or (NH4), ethanol/salt, and acetonitrile/D-glucose were used for partitioning of 503-antigen expressed in recombinant E. coli M15 with simultaneous LPSs removal. System based on 30% PEG 1500 and 10% potassium phosphate with 20% (w/w) unclarified cell homogenate achieved protein recovery of 116.96 ± 3.45% and purification factor of 1.55 ± 0.01. Endotoxin was removed with values as high as 99.90% for all systems with partial purification of 503-antigen, thus eliminating a clarification step in the partial purification protocol of this antigen.

Ethanol production by simultaneous saccharification and fermentation (SSF) using sugarcane bagasse as substrate was developed using batch and fed-batch mode. Acid, alkali, hydrothermal and hydrogen peroxide pretreatments to the sugarcane bagasse were tested. Experiments were carried out to optimize the enzyme load of cellulases and β-glucosidase. Four strains, two of Saccharomyces cerevisiae and two of Kluyveromyces marxianus yeast species were evaluate using SSF to produce ethanol. A kinetic study in bioreactor was carried out to optimize the SSF. The batch process was optimized using 1.0 g/L of inoculum, 15.0 FPU/g cellulose of cellulases and 6.0% of initial cellulose reaching 92.0% of theoretical ethanol yield after 18 h using the bagasse pretreate by acid-alkali and S. cerevisiae PE-2. The fed-batch process with enzyme load three times lower than that was used in batch process, obtained 88% of theoretical ethanol yield in 40 h. Therefore, the use of the lignocellulosic biomass (sugarcane bagasse) for producing a biofuel (ethanol) reduces the need for oil and is an environmental-friendly process.

Studies addressing for ecological compatible products have been increased along time, especially, on biosurfactant field. Biosurfactants are extracellular amphiphilic compound that are mainly produced by microorganisms and are classified into five main groups, including the glycolipids one. Rhamnolipids are included in the latter and are anionic biosurfactants produced predominantly by Pseudomonas aeruginosa being classified as mono- and di-rhamnolipids. In addition, their production may occur from different carbon sources, which may be obtained from renewable and low-cost residue. Therefore, it is possible to reduce the rhamnolipids production cost, since this has been the main bottleneck for replacing the chemical surfactants. In addition, to meeting a bona fide industrial application some limitations such as low productivity as well as recovery and/or purification that represent from 60 to 80% of total production cost should be improved. Therefore, this review covers different ways for producing rhamnolipids covering their application in many fields such as pharmaceutical, agricultural, petrochemical and so on; demonstrating the versatility of these biological compounds.

The present study aimed to evaluate different metals immobilized onto the Streamline™ chelating resin for purification of 503 antigen from Leishmania i. chagasi expressed in E. coli M15 by expanded...

Carnauba (Copernicia prunifera) is a Brazilian palm tree used for wax production, which usually generates a large amount of waste. This work evaluated the carnauba waste for cellulase and xylanase production using Trichoderma reesei CCT2768 through a solid-state fermentation (SSF). Carnauba waste was used in its crude form (C-IN), pretreated (C-P) with alkaline hydrogen peroxide (AHP), and also recycled after the SSF process (C-PR). C-IN, C-P, and C-PR were characterized by XRD, FTIR, and SEM. Cellulase and xylanase production was performed by SSF for 72 h, and the enzymatic extracts obtained were mixed each other in different concentrations. FPase, CMCase, and xylanase activities were determined. Trichoderma reesei CCT-2768 has shown high performance to produce cellulases and xylanases. Total cellulase, CMCase, and β-glycosidase presented a highest activity when C-PPR1 (25% of C-PR and 75% of C-P) was used as a carbon source, with yield of 2.85 U/g, 41.21 U/g, and 2.80 U/g, respectively. The highest xylanase production was achieved when only the pretreated carnauba waste (C-P) was used, with an enzyme activity of 224.93 U/g. Carnauba has shown a promising carbon source capacity to induce the production of cellulolytic and xylanolytic enzymes by using T. reesei CCT2768, promoting the circular and ecofriendly economy, as well as a cost reduction, of the production process of these enzymes.

The aim of the present study was to evaluate the effect of choline chloride-based deep eutectic solvents (DESs) as co-solvents for the ethanolic extraction of total carotenoids from buriti fruit (Mauritia flexuosa L.). Choline chloride-based DESs were synthesized by the heating method. The systems were prepared using ethanol and DESs as a co-solvent in concentrations of 0.5–70.0% (w/v). In addition, a control condition was performed using ethanol as the sole solvent. The total carotenoid concentration in the systems was determined by spectrophotometry at 450 nm. The results showed that the ethanol used without co-solvent obtained total carotenoid yields of 10.06 ± 0.03 mg/g for the buriti pulp and 10.43 ± 1.27 mg/g for the buriti peel. In turn, the use of choline chloride-based DESs as co-solvents did not increase the ethanolic extraction yield of carotenoids. However, it was observed that the buriti peel can be a rich source of carotenoids with yields comparable to the fruit. Thus, it is suggested that new green solvents be evaluated to increase the recovery of total carotenoids from buriti fruit using ecofriendly processes.