Lignocellulosic materials valorization in second generation biorefineries: an opportunity to produce fungal biopigments.

Second generation biorefineries play an important role in the production of renewable energy and fuels, utilizing forest and agro-industrial residues and by-products as raw materials. The integration of novel bioproducts, such as: xylitol, ß-carotene, xylooligosaccharides, and biopigments into the biorefinery's portfolio can offer economic benefits in the valorization of lignocellulosic materials, particularly cellulosic and hemicellulosic fractions. Fungal biopigments, known for their additional antioxidant and antimicrobial properties, are appealing to consumers and can have applications in various industrial sectors, including food and pharmaceuticals. The use of lignocellulosic materials as carbon and nutrient sources for the growth medium helps to reduce production costs, increasing the competitiveness of fungal biopigments in the market. In addition, the implementation of biopigment production in biorefineries allows the utilization of underutilized fractions, such as hemicellulose, for value-added bioproducts. This study deals with the potential of fungal biopigments production in second generation biorefineries in order to diversify the produced biomolecules together with energy generation. A comprehensive and critical review of the recent literature on this topic has been conducted, covering the major possible raw materials, general aspects of second generation biorefineries, the fungal biopigments and their potential for incorporation into biorefineries.

Using low-shear aerated and agitated bioreactor for producing two specific laccases by trametes versicolor cultures induced by 2,5-xylidine: Process development and economic analysis.

Laccase isoforms from basidiomycetes exhibit a superior redox potential compared to commercially available laccases obtained from ascomycete fungi, rendering them more reactive toward mono-substituted phenols and polyphenolic compounds. However, basidiomycetes present limitations for large-scale culture in liquid media, restraining the current availability of laccases from this fungal class. To advance laccase production from basidiomycetes, a newly designed 14-L low-shear aerated and agitated bioreactor provided enzyme titers up to 23.5 IU/mL from Trametes versicolor cultures. Produced enzymes underwent ultrafiltration and LC/MS-MS characterization, revealing the predominant production of only two out of the ten laccases predicted in the T. versicolor genome. Process simulation and economic analysis using SuperPro designer® suggested that T. versicolor laccase could be produced at US$ 3.60/kIU in a 200-L/batch enterprise with attractive economic parameters and a payback period of 1.7 years. The study indicates that new bioreactors with plain design help to produce low-cost enzymes from basidiomycetes.

Soil macrofauna communities in Brazilian land-use systems.

Background: Soil animal communities include more than 40 higher-order taxa, representing over 23% of all described species. These animals have a wide range of feeding sources and contribute to several important soil functions and ecosystem services. Although many studies have assessed macroinvertebrate communities in Brazil, few of them have been published in journals and even fewer have made the data openly available for consultation and further use. As part of ongoing efforts to synthesise the global soil macrofauna communities and to increase the amount of openly-accessible data in GBIF and other repositories related to soil biodiversity, the present paper provides links to 29 soil macroinvertebrate datasets covering 42 soil fauna taxa, collected in various land-use systems in Brazil. A total of 83,085 georeferenced occurrences of these taxa are presented, based on quantitative estimates performed using a standardised sampling method commonly adopted worldwide to collect soil macrofauna populations, i.e. the TSBF (Tropical Soil Biology and Fertility Programme) protocol. This consists of digging soil monoliths of 25 x 25 cm area, with handsorting of the macroinvertebrates visible to the naked eye from the surface litter and from within the soil, typically in the upper 0-20 cm layer (but sometimes shallower, i.e. top 0-10 cm or deeper to 0-40 cm, depending on the site). The land-use systems included anthropogenic sites managed with agricultural systems (e.g. pastures, annual and perennial crops, agroforestry), as well as planted forests and native vegetation located mostly in the southern Brazilian State of Paraná (96 sites), with a few additional sites in the neighbouring states of São Paulo (21 sites) and Santa Catarina (five sites). Important metadata on soil properties, particularly soil chemical parameters (mainly pH, C, P, Ca, K, Mg, Al contents, exchangeable acidity, Cation Exchange Capacity, Base Saturation and, infrequently, total N), particle size distribution (mainly % sand, silt and clay) and, infrequently, soil moisture and bulk density, as well as on human management practices (land use and vegetation cover) are provided. These data will be particularly useful for those interested in estimating land-use change impacts on soil biodiversity and its implications for below-ground foodwebs, ecosystem functioning and ecosystem service delivery. New information: Quantitative estimates are provided for 42 soil animal taxa, for two biodiversity hotspots: the Brazilian Atlantic Forest and Cerrado biomes. Data are provided at the individual monolith level, representing sampling events ranging from February 2001 up to September 2016 in 122 sampling sites and over 1800 samples, for a total of 83,085 ocurrences.

Fermentative Production of ß-Carotene from Sugarcane Bagasse Hydrolysate by Rhodotorula glutinis CCT-2186.

Β-Carotene is a red-orange pigment that serves as a precursor to important pharmaceutical molecules like vitamin A and retinol, making it highly significant in the industrial sector. Consequently, there is an ongoing quest for more sustainable production methods. In this study, glucose and xylose, two primary sugars derived from sugarcane bagasse (SCB), were utilized as substrates for ß-carotene production by Rhodotorula glutinis CCT-2186. To achieve this, SCB underwent pretreatment using NaOH, involved different concentrations of total solids (TS) (10%, 15%, and 20%) to remove lignin. Each sample was enzymatically hydrolyzed using two substrate loadings (5% and 10%). The pretreated SCB with 10%, 15%, and 20% TS exhibited glucose hydrolysis yields (%wt) of 93.10%, 91.88%, and 90.77%, respectively. The resulting hydrolysate was employed for ß-carotene production under batch fermentation. After 72 h of fermentation, the SCB hydrolysate yielded a ß-carotene concentration of 118.56 ± 3.01 mg/L. These findings showcase the robustness of R. glutinis as a biocatalyst for converting SCB into ß-carotene.

Microbial Biosurfactant: Candida bombicola as a Potential Remediator of Environments Contaminated by Heavy Metals.

Industrial interest in surfactants of microbial origin has intensified recently due to the characteristics of these compounds, such as biodegradability and reduced toxicity, and their efficiency in removing heavy metals and hydrophobic organic compounds from soils and waters. The aim of this study was to produce a biosurfactant using Candida bombicola URM 3712 in a low-cost medium containing 5.0% molasses, 3.0% corn steep liquor and 2.5% residual frying oil for 144 h at 200 rmp. Measurements of engine oil tension and emulsification were made under extreme conditions of temperature (0 °C, 5 °C, 70 °C, 100 °C and 120 °C), pH (2-12) and NaCl concentrations (2-12), demonstrating the stability of the biosurfactant. The isolated biosurfactant was characterized as an anionic molecule with the ability to reduce the surface tension of water from 72 to 29 mN/m, with a critical micellar concentration of 0.5%. The biosurfactant had no toxic effect on vegetable seeds or on Eisenia fetida as a bioindicator. Applications in the removal of heavy metals from contaminated soils under dynamic conditions demonstrated the potential of the crude and isolated biosurfactant in the removal of Fe, Zn and Pb with percentages between 70 and 88%, with the highest removal of Pb being 48%. The highest percentage of removal was obtained using the cell-free metabolic liquid, which was able to remove 48, 71 and 88% of lead, zinc and iron from the soil, respectively. Tests in packed columns also confirmed the biosurfactant's ability to remove Fe, Zn and Pb between 40 and 65%. The removal kinetics demonstrated an increasing percentage, reaching removal of 50, 70 and 85% for Pb, Zn and Fe, respectively, reaching a greater removal efficiency at the end of 24 h. The biosurfactant was also able to significantly reduce the electrical conductivity of solutions containing heavy metals. The biosurfactant produced by Candida bombicola has potential as an adjuvant in industrial processes for remediating soils and effluents polluted by inorganic contaminants.

Surfactants, Biosurfactants, and Non-Catalytic Proteins as Key Molecules to Enhance Enzymatic Hydrolysis of Lignocellulosic Biomass.

Lignocellulosic biomass (LCB) has remained a latent alternative resource to be the main substitute for oil and its derivatives in a biorefinery concept. However, its complex structure and the underdeveloped technologies for its large-scale processing keep it in a state of constant study trying to establish a consolidated process. In intensive processes, enzymes have been shown to be important molecules for the fractionation and conversion of LCB into biofuels and high-value-added molecules. However, operational challenges must be overcome before enzyme technology can be the main resource for obtaining second-generation sugars. The use of additives is shown to be a suitable strategy to improve the saccharification process. This review describes the mechanisms, roles, and effects of using additives, such as surfactants, biosurfactants, and non-catalytic proteins, separately and integrated into the enzymatic hydrolysis process of lignocellulosic biomass. In doing so, it provides a technical background in which operational biomass processing hurdles such as solids and enzymatic loadings, pretreatment burdens, and the unproductive adsorption phenomenon can be addressed.

High-solid enzymatic hydrolysis of sugarcane bagasse and ethanol production in repeated batch process using column reactors.

Alkaline sulfite pretreated sugarcane bagasse was enzymatically hydrolyzed in a packed-bed column reactor and a bubble column reactor was evaluated to produce ethanol from the hydrolysate. Initial solid loadings of 9-16% were used in column reactor in the hydrolysis step, and the use of lower value (9%) resulted in 41 g L-1 of glucose in the hydrolysate, corresponding to 87% of cellulose hydrolysis yield. This yield was reduced to 65% for a solid loading of 16%, corresponding to a glucose concentration of 54 g L-1. Subsequently, Saccharomyces cerevisiae and Scheffersomyces stipitis were used for ethanol production in medium based on hydrolysate previously obtained, using different aeration flowrates (0.3, 0.5 and 0.7 vvm). In simple batch fermentation using S. cerevisiae, higher ethanol yield (0.40 g.g-1) and productivity (1.58 g.L-1.h-1) were achieved using 0.5 vvm. When S. stipitis was used in simple batch co-fermentations, the maximum ethanol productivities were obtained using 0.5 and 0.7 vvm (0.64 and 0.63 g.L-1.h-1, respectively). Successive repeated batches resulted in average ethanol concentration of 38 g.L-1 and fermentation efficiency of 82%, when using S. cerevisiae. For S. stipitis, those values were, respectively, 36 g.L-1 and 50%, with volumetric productivity increased along the cycles. Thus, the potential of the bioreactors as simple systems for use in the biological steps of biorefineries was demonstrated. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02932-3.

Chemical Modification of Pullulan Exopolysaccharide by Grafting Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBHV) via Click Chemistry.

Biodegradable and biocompatible copolymers have been often studied for the development of biomaterials for drug delivery systems. In this context, this work reports the synthesis and characterization of a novel pullulan-g-poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (Pull-g-PHBHV) graft copolymer using click chemistry. Well-defined and functional pullulan backbones containing azide groups (PullN3) previously prepared by our group were successfully used for this purpose and propargyl-terminated poly(3-hydroxybutyrate-co-3-hydroxyvalerate) was prepared via transesterification using propargyl alcohol as a chain transfer agent. By an alkyne-azide cycloaddition reaction catalyzed by copper (Cu (I)) (CuAAC), the graft copolymer Pull-g-PHBHV was obtained. The chemical structures of the polymers were accessed by 1H NMR and 13C NMR FTIR. Disappearance of the bands referring to the main bonds evidenced success in the grafting reaction. Besides that, DRX, DSC and TGA were used in order to access the changes in crystallinity and thermal behavior of the material. The remaining crystallinity of the Pull-g-PHBHV structure evidences the presence of PHBHV. Pull-g-PHBHV presented lower degradation maximum temperature values than the starting materials, indicating its minor thermal stability. Finally, the synthesized material is an innovative biopolymer, which has never been reported in the previous literature. It is a bio-derived and biodegradable polymer, chemically modified, resulting in interesting properties which can be useful for their further applications as biomedical systems for controlled delivery, for example.

Synthesis of amphiphilic pullulan-graft-poly(ε-caprolactone) via click chemistry.

Chemical modification of natural polymers has been commonly employed for the development of new bio-based materials, aiming at adjusting specific properties such as solubility, biodegradability, thermal stability and mechanical behavior. Among all natural polymers, polysaccharides are promising materials, in which biodegradability, processability and bioreactivity make them suitable for biomedical applications. In this context, this work describes the synthesis and characterization of a novel amphiphilic pullulan-g-poly(ε-caprolactone) (Pull-g-PCL) graft copolymer. In a first step, pullulan was chemically modified with 2-bromopropionyl bromide to obtain bromo-functionalized pullulan (PullBr). Then, this precursor was modified with sodium azide, leading to azide pullulan (PullN3). In parallel, propargyl-terminated poly(ε-caprolactone) was prepared via ring-opening polymerization (ROP). These preliminary steps involved the synthesis of azide and alkyne compounds, capable of being linked together via alkyne-azide cycloaddition reaction catalyzed by copper (Cu (I)), which leads to Pull-g-PCL. The chemical structures of the polymers were assessed by Proton Nuclear Magnetic Resonance (1H NMR) and Fourier Transform Infrared (FTIR).

Theoretical Thermodynamics Study of Polyamidoamine Deposited Around a Nanotube as Motor Controlled by Light and Under Temperature Effect.

We simulated a system like a Polyamidoamine (PAMAM) deposited on open carbon nanotube. We used five first generation PAMAM. The initial position of PAMAM is out of CN symmetry position. It permits the PAMAM to relax around the nanotube due to van der Waals force. After that, we have analyzed the thermal effects on behavior of 4G PAMAM. We performed computational simulation using classical molecular dynamics with standard parameterization. The thermodynamics properties of this device as molar specific heat and molar entropy variation were calculated. The CN has 690 carbon atoms with up to almost 0.1 ns of simulation.

Molecular dynamics study of the flagella inside the carbon torus.

We propose a system that avoids the open ends of the nanotube, we used a device consisting of flagellum (FLA) inside a nanotorus (NT) formed by carbon atoms. The flagellum consists of a C20 nanosphere with fixed size of tail within the NT. The full system consists of a closed loop drive and static nanotubes and nanospheres not static with different sizes of flagella released inside the nanotube, with each simulation, allows the relaxation between (internal and external NT). The nanospheres result in a system that provides movement of Van der Waals. The simulations were done by well-known classic molecular dynamics with standard parameterization. We calculate thermodynamic properties of these devices as heat capacity and molar entropy variation. For this system were obtained properties such as: the speed of nanospheres plagued the efficiency of molecular motor versus time, the kinetic energy, potential energy and total energy in each of the simulations. In our calculations, this system has a number of carbon atoms ranging from (2721 until 2728) with up to almost 10 ps simulation. These facts can be useful for the construction of new molecular machines.

Screening of lipases for the synthesis of xylitol monoesters by chemoenzymatic esterification and the potential of microwave and ultrasound irradiations to enhance the reaction rate.

Lipases from different sources, Pseudomonas fluorescens (AK lipase), Burkholderia cepacia (PS lipase), Penicillium camembertii (lipase G) and Porcine pancreas lipase (PPL), previously immobilized on epoxy SiO(2)-PVA, were screened for the synthesis of xylitol monoesters by esterification of the protected xylitol using oleic acid as acyl donor group. Among all immobilized derivatives, the highest esterification yield was achieved by P. camembertii lipase, showing to be attractive alternative to bulk chemical routes to satisfy increasing commercial demands. Further experiments were performed to determine the influence of fatty acids chain size on the reaction yield and the feasibility of using non-conventional heating systems (microwave and ultrasound irradiations) to enhance the reaction rate.

Evaluation of the catalytic properties of Burkholderia cepacia lipase immobilized on non-commercial matrices to be used in biodiesel synthesis from different feedstocks.

The objective of this work was to produce an immobilized form of lipase from Burkholderia cepacia (lipase PS) with advantageous catalytic properties and stability to be used in the ethanolysis of different feedstocks, mainly babassu oil and tallow beef. For this purpose lipase PS was immobilized on two different non-commercial matrices, such as inorganic matrix (niobium oxide, Nb(2)O(5)) and a hybrid matrix (polysiloxane-polyvinyl alcohol, SiO(2)-PVA) by covalent binding. The properties of free and immobilized enzymes were searched and compared. The best performance regarding all the analyzed parameters (biochemical properties, kinetic constants and thermal stability) were obtained when the lipase was immobilized on SiO(2)-PVA. The superiority of this immobilized system was also confirmed in the transesterification of both feedstocks, attained higher yields and productivities.

Packed-bed reactor running on babassu oil and glycerol to produce monoglycerides by enzymatic route using immobilized Burkholderia cepacia lipase.

The aim of this study was the glycerolysis of babassu oil catalyzed by immobilized lipase from Burkholderia cepacia, in a continuous packed-bed reactor. The best reaction conditions were previously established in batchwise via response surface methodology as a function of glycerol-to-oil molar ratio and reaction temperature. The reactor operated continuously for 22 days at 50 degrees C, and during the first 6 days, no significant decrease on the initial lipase activity was observed. Monoglycerides concentration was in the range from 25 to 33 wt.%. Subsequently, a progressive decrease in the activity was detected, and an inactivation profile described by Arrhenius model estimated values of 50 days and 1.37 x 10(-2) h(-1) the half-life and deactivation coefficient, respectively.

PVA-hydrogel entrapped Candida guilliermondii for xylitol production from sugarcane hemicellulose hydrolysate.

Viable cells of Candida guilliermondii were immobilized by inclusion into polyvinyl alcohol (PVA) hydrogel using the freezing-thawing method. Entrapment experiments were planned according to a 2(3) full factorial design, using the PVA concentration (80, 100, and 120 g L(-1)), the freezing temperature (-10, -15, and -20 degrees C), and the number of freezing-thawing cycles (one, three, and five) as the independent variables, integrated with three additional tests to estimate the errors. The effectiveness of the immobilization procedure was checked in Erlenmeyer flasks as the pellet capability to catalyze the xylose-to-xylitol bioconversion of a medium based on sugarcane bagasse hemicellulosic hydrolysate. To this purpose, the yield of xylitol on consumed xylose, xylitol volumetric productivity, and cell retention yield were selected as the response variables. Cell pellets were then used to perform the same bioconversion in a stirred tank reactor operated at 400 rpm, 30 degrees C, and 1.04 vvm air flowrate. At the end of fermentation, a maximum xylitol concentration of 28.7 g L(-1), a xylitol yield on consumed xylose of 0.49 g g(-1) and a xylitol volumetric productivity of 0.24 g L(-1) h(-1) were obtained.

Total soluble solids from banana: evaluation and optimization of extraction parameters.

Banana, an important component in the diet of the global population, is one of the most consumed fruits in the world. This fruit is also very favorable to industry processes (e.g., fermented beverages) due to its rich content on soluble solids and minerals, with low acidity. The main objective of this work was to evaluate the influence of factors such as banana weight and extraction time during a hot aqueous extraction process on the total soluble solids content of banana. The extract is to be used by the food and beverage industries. The experiments were performed with 105 mL of water, considering the moisture of the ripe banana (65%). Total sugar concentrations were obtained in a beer analyzer and the result expressed in degrees Plato (degrees P, which is the weight of the extract or the sugar equivalent in 100 g solution at 20 degrees C), aiming at facilitating the use of these results by the beverage industries. After previous studies of characterization of the fruit and of ripening performance, a 2(2) full-factorial star design was carried out, and a model was developed to describe the behavior of the dependent variable (total soluble solids) as a function of the factors (banana weight and extraction time), indicating as optimum conditions for extraction 38.5 g of banana at 39.7 min.

Covalent attachment of Candida rugosa lipase on chemically modified hybrid matrix of polysiloxane-polyvinyl alcohol with different activating compounds.

Candida rugosa lipase was immobilized by covalent binding on hybrid matrix of polysiloxane-polyvinyl alcohol chemically modified with different activating agents as glutaraldehyde, sodium metaperiodate and carbonyldiimidazole. The experimental results suggested that functional activating agents render different interactions between enzyme and support, producing consequently alterations in the optimal reaction conditions. Properties of the immobilized systems were assessed and their performance on hydrolytic and synthetic reactions were evaluated and compared with the free enzyme. In hydrolytic reactions using p-nitrophenyl palmitate as substrate all immobilized systems showed higher thermal stability and optima pH and temperature values in relation to the free lipase. Among the activating compounds, carbonyldiimidazole resulted in a total recovery of activity on the support and the highest thermal stability. For the butyl butyrate synthesis, the best performance (molar conversion of 95% and volumetric productivity of 2.33 g L(-1)h(-1)) was attained with the lipase immobilized on POS-PVA activated with sodium metaperiodate. The properties of the support and immobilized derivatives were also evaluated by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopies and chemical composition (FTIR).

Cell immobilization and xylitol production using sugarcane bagasse as raw material.

Sugarcane bagasse pretreated by three different procedures (with 2% [v/v] polyethyleneimine (PEI), with 2% [w/v] NaOH, or with a sequence of NaOH and PEI) was used as cell immobilization carrier for xylitol production by Candida guilliermondii yeast. Fermentations using these pretreated carriers were performed in semidefined medium and in a hydrolysate medium produced from sugarcane bagasse hemicellulose. Sugarcane bagasse pretreated with NaOH was the best carrier obtained with respect to immobilization efficiency, because it was able to immobilize a major quantity of cells (0.30 g of cells/g of bagasse). Fermentation in semidefined medium using the NaOH-pretreated carrier attained a high efficiency of xylose-to-xylitol bioconversion (96% of the theoretical value). From hydrolysate medium, the bioconversion efficiency was lower (63%), probably owing to the presence of other substances in the medium that caused an inadequate mass transfer to the cells. In this fermentation medium, better results with relation to xylitol production were obtained by using PEI-pretreated carrier (xylose-to-xylitol bioconversion of 81% of the theoretical and volumetric productivity of 0.43 g/[L.h]). The results showed that sugarcane bagasse is a low-cost material with great potential for use as cell immobilization carrier in the fermentative process for xylitol production.

Enzymatic hydrolysis of soybean oil using lipase from different sources to yield concentrated of polyunsaturated fatty acids.

The ability of three commercially available lipases to mediate the hydrolysis of the soybean oil to yield concentrated of essential fatty acids was evaluated. The tested lipases were from microbial (Candida rugosa and Thermomyces lanuginosa) and animal cells (Porcine pancreatic lipase). In terms of free fatty acids, microbial lipases were more effective to promote the enzymatic hydrolysis of the soybean oil (over 70%) than the porcine pancreatic lipase (24%). In spite of this, porcine pancreatic lipase (PPL) showed the most satisfactory specificity towards both essential fatty acids and was, therefore, chosen to carry out additional studies. An experimental design was performed taking into consideration the enzyme and NaCl amounts as independent variables. The main effects were fitted by multiple regression analysis to a linear model and maximum fatty acids concentration could be obtained using 3.0 wt% of lipase and 0.08 wt% of NaCl. The mathematical model representing the hydrolysis degree was found to describe adequately the experimental results. Under these conditions, concentrations of 29.5 g/L and 4.6 g/L for linoleic and linolenic acids, respectively, were obtained.

Variables that affect xylitol production from sugarcane bagasse hydrolysate in a zeolite fluidized bed reactor.

The operational conditions for xylitol production by fermentation of sugarcane bagasse hydrolysate in a fluidized bed reactor with cells immobilized on zeolite were evaluated. Fermentations were carried out under different conditions of air flowrate (0.0125-0.0375 vvm), zeolite mass (100-200 g), initial pH (4-6), and xylose concentration (40-60 g/L), according to a 2(4) full factorial design. The air flowrate increase resulted in a metabolic deviation from product to biomass formation. On the other hand, the pH increase favored both the xylitol yield (Y(P/S)) and volumetric productivity (Q(P)), and the xylose concentration increase positively influenced the xylitol concentration. The best operational conditions evaluated were based on the use of an air flowrate of 0.0125 vvm, 100 g of zeolite, pH 6, and xylose concentration of 60 g/L. Under these conditions, 38.5 g/L of xylitol were obtained, with a Y(P/S) of 0.72 g/g, Q(P) of 0.32 g/L.h, and cell retention of 25.9%.