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1.
Sci Rep ; 13(1): 19182, 2023 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-37932303

RESUMO

Simultaneous intracellular depolymerization of xylo-oligosaccharides (XOS) and acetate fermentation by engineered Saccharomyces cerevisiae offers significant potential for more cost-effective second-generation (2G) ethanol production. In the present work, the previously engineered S. cerevisiae strain, SR8A6S3, expressing enzymes for xylose assimilation along with an optimized route for acetate reduction, was used as the host for expressing two ß-xylosidases, GH43-2 and GH43-7, and a xylodextrin transporter, CDT-2, from Neurospora crassa, yielding the engineered SR8A6S3-CDT-2-GH34-2/7 strain. Both ß-xylosidases and the transporter were introduced by replacing two endogenous genes, GRE3 and SOR1, that encode aldose reductase and sorbitol (xylitol) dehydrogenase, respectively, and catalyse steps in xylitol production. The engineered strain, SR8A6S3-CDT-2-GH34-2/7 (sor1Δ gre3Δ), produced ethanol through simultaneous XOS, xylose, and acetate co-utilization. The mutant strain produced 60% more ethanol and 12% less xylitol than the control strain when a hemicellulosic hydrolysate was used as a mono- and oligosaccharide source. Similarly, the ethanol yield was 84% higher for the engineered strain using hydrolysed xylan, compared with the parental strain. Xylan, a common polysaccharide in lignocellulosic residues, enables recombinant strains to outcompete contaminants in fermentation tanks, as XOS transport and breakdown occur intracellularly. Furthermore, acetic acid is a ubiquitous toxic component in lignocellulosic hydrolysates, deriving from hemicellulose and lignin breakdown. Therefore, the consumption of XOS, xylose, and acetate expands the capabilities of S. cerevisiae for utilization of all of the carbohydrate in lignocellulose, potentially increasing the efficiency of 2G biofuel production.


Assuntos
Saccharomyces cerevisiae , Xilosidases , Saccharomyces cerevisiae/metabolismo , Xilanos/metabolismo , Xilose/metabolismo , Etanol/metabolismo , Engenharia Metabólica , Xilitol/metabolismo , Oligossacarídeos/metabolismo , Fermentação , D-Xilulose Redutase/genética , D-Xilulose Redutase/metabolismo , Xilosidases/metabolismo , Acetatos/metabolismo
2.
Rev. Inst. Adolfo Lutz ; 82: e38777, maio 2023. graf, tab
Artigo em Português | LILACS, CONASS, Coleciona SUS, Sec. Est. Saúde SP, VETINDEX, SESSP-ACVSES, SESSP-IALPROD, Sec. Est. Saúde SP | ID: biblio-1435644

RESUMO

Xilooligossacarídeos (XOS) são reconhecidos pelo seu potencial prebiótico relevante para diversos setores industriais e foram obtidos após o pré-tratamento hidrotérmico da biomassa lignocelulósica residual de galhos de eucalipto. Subprodutos inibitórios são gerados durante o processo de solubilização dos oligossacarídeos e acabam comprometendo a utilização do licor em microrganismos. Neste trabalho, o processo de destoxificação, hidrólise enzimática e atividade estimulantes de crescimento da bactéria Staphylococcus xylosus foram estabelecidos. Os resultados mostraram que a adsorção com carvão ativado em pó removeu cerca de 55% do ácido acético e mais de 90% do ácido fórmico, compostos fenólicos, lignina solúvel, furfural e 5-hidroximetilfurfural, e que a soma dos oligossacarídeos xilobiose (X2) e xilotriose (X3) foram maximizadas de 0,57 g/L para 1,21 g/L com 110 U/gXOS da enzima endoxilanase e 6,3% do licor destoxificado na hidrólise enzimática. O consumo de cerca de 63% de X2 e de 46% de X3 pela bactéria em meio basal deficiente em fontes de carbono, mas acrescido com os oligômeros, proporcionou maior crescimento celular em relação aos meios basais com alta composição de carbono, com e sem XOS, revelando seu potencial prebiótico pelo efeito estimulante de crescimento. (AU)


Xylooligosaccharides (XOS) are recognized for their prebiotic potential relevant to several industrial sectors and were obtained after hydrothermal pretreatment of residual lignocellulosic biomass from eucalyptus branches. Inhibitory by-products are generated during the solubilization process of oligosaccharides and end up compromising the utilization of the liquor in microorganisms. In this work, the detoxification process, enzymatic hydrolysis and growth stimulating activity of Staphylococcus xylosus bacteria were established. The results showed that adsorption with powdered activated carbon removed about 55% of acetic acid and more than 90% of formic acid, phenolic compounds, soluble lignin, furfural, and 5-hydroxymethyl furfural and the sum of the oligosaccharides xylobiose (X2) and xylotriose (X3) were maximized from 0.57 g/L to 1.21 g/L with 110 U/gXOS of the enzyme endoxylanase and 6.3% of the detoxified liquor in the enzymatic hydrolysis. The consumption of X2 and X3 were about 63% and 46%, respectively, by the bacteria in basal medium deficient in carbon sources, but in medium added with the oligomers, provided higher cell growth compared to basal medium with high carbon composition, with and without XOS, revealing its prebiotic potential by its growth-stimulating effect. (AU)


Assuntos
Oligossacarídeos , Staphylococcus , Xilose , Carvão Vegetal , Biomassa , Eucalyptus , Prebióticos
3.
Microbiol Spectr ; 10(3): e0212521, 2022 06 29.
Artigo em Inglês | MEDLINE | ID: mdl-35658600

RESUMO

Lytic polysaccharide monooxygenases (LPMOs) are oxidative enzymes found in viruses, archaea, and bacteria as well as eukaryotes, such as fungi, algae and insects, actively contributing to the degradation of different polysaccharides. In Aspergillus nidulans, LPMOs from family AA9 (AnLPMO9s), along with an AA3 cellobiose dehydrogenase (AnCDH1), are cosecreted upon growth on crystalline cellulose and lignocellulosic substrates, indicating their role in the degradation of plant cell wall components. Functional analysis revealed that three target LPMO9s (AnLPMO9C, AnLPMO9F and AnLPMO9G) correspond to cellulose-active enzymes with distinct regioselectivity and activity on cellulose with different proportions of crystalline and amorphous regions. AnLPMO9s deletion and overexpression studies corroborate functional data. The abundantly secreted AnLPMO9F is a major component of the extracellular cellulolytic system, while AnLPMO9G was less abundant and constantly secreted, and acts preferentially on crystalline regions of cellulose, uniquely displaying activity on highly crystalline algae cellulose. Single or double deletion of AnLPMO9s resulted in about 25% reduction in fungal growth on sugarcane straw but not on Avicel, demonstrating the contribution of LPMO9s for the saprophytic fungal lifestyle relies on the degradation of complex lignocellulosic substrates. Although the deletion of AnCDH1 slightly reduced the cellulolytic activity, it did not affect fungal growth indicating the existence of alternative electron donors to LPMOs. Additionally, double or triple knockouts of these enzymes had no accumulative deleterious effect on the cellulolytic activity nor on fungal growth, regardless of the deleted gene. Overexpression of AnLPMO9s in a cellulose-induced secretome background confirmed the importance and applicability of AnLPMO9G to improve lignocellulose saccharification. IMPORTANCE Fungal lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that boost plant biomass degradation in combination with glycoside hydrolases. Secretion of LPMO9s arsenal by Aspergillus nidulans is influenced by the substrate and time of induction. These findings along with the biochemical characterization of novel fungal LPMO9s have implications on our understanding of their concerted action, allowing rational engineering of fungal strains for biotechnological applications such as plant biomass degradation. Additionally, the role of oxidative players in fungal growth on plant biomass was evaluated by deletion and overexpression experiments using a model fungal system.


Assuntos
Aspergillus nidulans , Oxigenases de Função Mista , Aspergillus nidulans/genética , Celulose/química , Celulose/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Lignina , Oxigenases de Função Mista/genética , Oxigenases de Função Mista/metabolismo , Polissacarídeos , Secretoma
4.
Bioresour Technol ; 357: 127093, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35378280

RESUMO

This study investigated the production of xylo-oligosaccharides (XOS) from sugarcane straw (SCS) using steam explosion (SE) pretreatment at pilot-scale, as well as co-production of fermentable sugars and lignin-rich residues for bioethanol and bioenergy, respectively. SE conditions 200 °C; 15 bar; 10 min led to 1) soluble XOS yields of up to 35 % (w/w) of initial xylan with âˆ¼50 % of the recovered XOS corresponding to xylobiose and xylotriose, considered the most valuable sugars for prebiotic applications; 2) fermentable glucose yields from the enzymatic hydrolysis of SE-pretreated SCS of up to âˆ¼78 %; 3) increase in the energy content of saccharified SCS residues (16 %) compared to the untreated material. From an integrated biorefinery perspective, it demonstrated the potential use of SCS for the production of value-added XOS ingredients as well as liquid and solid biofuel products.


Assuntos
Saccharum , Grão Comestível , Hidrólise , Oligossacarídeos , Vapor , Açúcares
5.
Front Bioeng Biotechnol ; 10: 825981, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35242749

RESUMO

The engineering of xylo-oligosaccharide-consuming Saccharomyces cerevisiae strains is a promising approach for more effective utilization of lignocellulosic biomass and the development of economic industrial fermentation processes. Extending the sugar consumption range without catabolite repression by including the metabolism of oligomers instead of only monomers would significantly improve second-generation ethanol production This review focuses on different aspects of the action mechanisms of xylan-degrading enzymes from bacteria and fungi, and their insertion in S. cerevisiae strains to obtain microbial cell factories able of consume these complex sugars and convert them to ethanol. Emphasis is given to different strategies for ethanol production from both extracellular and intracellular xylo-oligosaccharide utilization by S. cerevisiae strains. The suitability of S. cerevisiae for ethanol production combined with its genetic tractability indicates that it can play an important role in xylan bioconversion through the heterologous expression of xylanases from other microorganisms.

6.
Appl Microbiol Biotechnol ; 105(23): 8969-8987, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34698899

RESUMO

This work performed co-AD from the vinasse and filter cake (from 1G ethanol production) and deacetylation liquor (from the pretreatment of sugarcane straw for 2G ethanol production) in a semi-Continuous Stirred Tank Reactor (s-CSTR) aiming to provide optimum operational parameters for continuous CH4 production. Using filter cake as co-substrate may allow the reactor to operate throughout the year, as it is available in the sugarcane off-season, unlike vinasse. A comparison was made from the microbial community of the seed sludge and the reactor sludge when CH4 production stabilized. Lactate, butyrate, and propionate fermentation routes were denoted at the start-up of the s-CSTR, characterizing the acidogenic phase: the oxidation-reduction potential (ORP) values ranged from -800 to -100 mV. Once the methanogenesis was initiated, alkalizing addition was no longer needed as its demand by the microorganisms was supplied by the alkali characteristics of the deacetylation liquor. The gradual increase of the applied organic load rates (OLR) allowed stabilization of the methanogenesis from 3.20 gVS L-1 day-1: the highest CH4 yield (230 mLNCH4 g-1VS) and average organic matter removal efficiency (83% ± 13) was achieved at ORL of 4.16 gVS L-1 day-1. The microbial community changed along with the reactor operation, presenting different metabolic routes mainly due to the used lignocellulosic substrates. Bacteria from the syntrophic acetate oxidation (SAO) process coupled to hydrogenotrophic methanogenesis were predominant (~ 90% Methanoculleus) during the CH4 production stability. The overall results are useful as preliminary drivers in terms of visualizing the co-AD process in a sugarcane biorefinery integrated to scale. KEY POINTS: • Integration of 1G2G sugarcane ethanol biorefinery from co-digestion of its residues. • Biogas production from vinasse, filter cake, and deacetylation liquor in a semi-CSTR. • Lignocellulosic substrates affected the biochemical routes and microbial community. • Biomol confirmed the establishment of the thermophilic community from mesophilic sludge.


Assuntos
Saccharum , Anaerobiose , Biocombustíveis , Reatores Biológicos , Digestão , Metano
7.
Front Oncol ; 11: 639339, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34026616

RESUMO

Cervical cancer (CC) represents a major global health issue, particularly impacting women from resource constrained regions worldwide. Treatment refractoriness to standard chemoradiotheraphy has identified cancer stem cells as critical coordinators behind the biological mechanisms of resistance, contributing to CC recurrence. In this work, we evaluated differential gene expression in cervical cancer stem-like cells (CCSC) as biomarkers related to intrinsic chemoradioresistance in CC. A total of 31 patients with locally advanced CC and referred to Mário Penna Institute (Belo Horizonte, Brazil) from August 2017 to May 2018 were recruited for the study. Fluorescence-activated cell sorting was used to enrich CD34+/CD45- CCSC from tumor biopsies. Transcriptome was performed using ultra-low input RNA sequencing and differentially expressed genes (DEGs) using Log2 fold differences and adjusted p-value < 0.05 were determined. The analysis returned 1050 DEGs when comparing the Non-Responder (NR) (n=10) and Responder (R) (n=21) groups to chemoradiotherapy. These included a wide-ranging pattern of underexpressed coding genes in the NR vs. R patients and a panel of lncRNAs and miRNAs with implications for CC tumorigenesis. A panel of biomarkers was selected using the rank-based AUC (Area Under the ROC Curve) and pAUC (partial AUC) measurements for diagnostic sensitivity and specificity. Genes overlapping between the 21 highest AUC and pAUC loci revealed seven genes with a strong capacity for identifying NR vs. R patients (ILF2, RBM22P2, ACO16722.1, AL360175.1 and AC092354.1), of which four also returned significant survival Hazard Ratios. This study identifies DEG signatures that provide potential biomarkers in CC prognosis and treatment outcome, as well as identifies potential alternative targets for cancer therapy.

8.
Biotechnol Biofuels ; 14(1): 96, 2021 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-33865436

RESUMO

BACKGROUND: Wood-decay basidiomycetes are effective for the degradation of highly lignified and recalcitrant plant substrates. The degradation of lignocellulosic materials by brown-rot strains is carried out by carbohydrate-active enzymes and non-enzymatic Fenton mechanism. Differences in the lignocellulose catabolism among closely related brown rots are not completely understood. Here, a multi-omics approach provided a global understanding of the strategies employed by L. sulphureus ATCC 52600 for lignocellulose degradation. RESULTS: The genome of Laetiporus sulphureus ATCC 52600 was sequenced and phylogenomic analysis supported monophyletic clades for the Order Polyporales and classification of this species within the family Laetiporaceae. Additionally, the plasticity of its metabolism was revealed in growth analysis on mono- and disaccharides, and polysaccharides such as cellulose, hemicelluloses, and polygalacturonic acid. The response of this fungus to the presence of lignocellulosic substrates was analyzed by transcriptomics and proteomics and evidenced the occurrence of an integrated oxidative-hydrolytic metabolism. The transcriptomic profile in response to a short cultivation period on sugarcane bagasse revealed 125 upregulated transcripts, which included CAZymes (redox enzymes and hemicellulases) as well as non-CAZy redox enzymes and genes related to the synthesis of low-molecular-weight compounds. The exoproteome produced in response to extended cultivation time on Avicel, and steam-exploded sugarcane bagasse, sugarcane straw, and Eucalyptus revealed 112 proteins. Contrasting with the mainly oxidative profile observed in the transcriptome, the secretomes showed a diverse hydrolytic repertoire including constitutive cellulases and hemicellulases, in addition to 19 upregulated CAZymes. The secretome induced for 7 days on sugarcane bagasse, representative of the late response, was applied in the saccharification of hydrothermally pretreated grass (sugarcane straw) and softwood (pine) by supplementing a commercial cocktail. CONCLUSION: This study shows the singularity of L. sulphureus ATCC 52600 compared to other Polyporales brown rots, regarding the presence of cellobiohydrolase and peroxidase class II. The multi-omics analysis reinforces the oxidative-hydrolytic metabolism involved in lignocellulose deconstruction, providing insights into the overall mechanisms as well as specific proteins of each step.

9.
Bioresour Technol ; 323: 124625, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33418350

RESUMO

Pretreatment strategies are fundamental to effectively deconstruct lignocellulosic biomass and economically produce biofuels, biomaterials and bio-based chemicals. This study evaluated individual and combinatorial steam explosion (SE) and ionic liquid (IL) pretreatments for production of high-value oligosaccharides from a novel seed-based Miscanthus hybrid (Mx2779). The two ILs used for pretreatment were triethylammonium hydrogen sulphate [TEA][HSO4] and 1-ethyl-3-methylimidazolium acetate [C2mim][OAc]. The results showed that each pretreatment leads to distinct effects on the fragmentation (cellulose and xylan dissolution, delignification, deacetylation) and physicochemical modification (cellulose and lignin properties) of lignocellulose. This, in turn, dictated enzymatic hydrolysis efficiencies of the cellulose pulp to glucose or gluco-oligosaccharides for downstream applications. Our findings suggest that the stand-alone SE or [C2mim][OAc] pretreatments may offer cost advantages over [TEA][HSO4] through the production of oligosaccharides such as xylo- and gluco-oligosaccharides. This study also highlights technical and economic pretreatment process challenges related to the production of oligosaccharides from Miscanthus Mx2779 biomass.


Assuntos
Biocombustíveis , Líquidos Iônicos , Biomassa , Hidrólise , Lignina , Oligossacarídeos , Vapor
10.
Biotechnol Lett ; 43(5): 967-979, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33517513

RESUMO

OBJECTIVE: The extraction of the hemicellulose fraction of sugarcane bagasse (SCB) by acid hydrolysis was evaluated in an autoclave and a Parr reactor aiming the application of the hydrolysate as a carbon source for lipid production by Lipomyces starkeyi. RESULTS: The hydrolysis that resulted in the highest sugar concentration was obtained by treatment in the Parr reactor (HHR) at 1.5% (m/v) H2SO4 and 120 °C for 20 min, reaching a hemicellulose conversion of approximately 82%. The adaptation of the yeast to the hydrolysate provided good fermentability and no lag phase. The fermentation of hemicellulose-derived sugars (HHR) by L. starkeyi resulted in a 27.8% (w/w) lipid content and YP/S of 0.16 g/l.h. Increasing the inoculum size increased the lipid content by approximately 61%, reaching 44.8% (w/w). CONCLUSION: The hemicellulose hydrolysate from SCB is a potential substrate for L. starkeyi to produce lipids for biodiesel synthesis based on the biorefinery concept.


Assuntos
Lipomyces/metabolismo , Óleos/metabolismo , Polissacarídeos/química , Saccharum/química , Adaptação Fisiológica , Biocombustíveis , Reatores Biológicos , Celulose/química , Celulose/metabolismo , Fermentação , Temperatura Alta , Hidrólise , Lipídeos/biossíntese , Lipomyces/crescimento & desenvolvimento , Polissacarídeos/metabolismo , Açúcares/química , Açúcares/metabolismo , Ácidos Sulfúricos/química
11.
Bioresour Technol ; 316: 123918, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32763802

RESUMO

Cello-oligosaccharides (COS) are oligomers with 2 to 6 ß-1,4-linked glucose units, with potential applications in the food/feed and bioenergy industrial sectors. In this study, the combination of five heterologous expressed endoglucanases varying the temperature and pH conditions were evaluated by design of experiments for COS production. Afterwards, the best combination was tested to produce COS from different pretreated sugarcane straws: ionic liquid, diluted acid, hydrothermal and steam-explosion. The results showed that steam explosion pretreated sugarcane straw treated with CtCel9R enzyme at 50 °C and pH 5.0 yielded 13.4 mg COS g biomass-1, 5-18-fold higher compared to the other pretreated straws. Under the conditions evaluated, the removal of hemicellulose and decrease in the cellulose crystallinity can benefits the enzymatic hydrolysis. This is the first study that combined the evaluation of different enzymes, conditions, and sugarcane straw pretreatments to optimize COS production in a single step without glucose formation.


Assuntos
Celulase , Saccharum , Celulose , Hidrólise , Oligossacarídeos
12.
Bioresour Technol ; 313: 123637, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32535521

RESUMO

Sugarcane straw (SS) is a widely available agricultural processing feedstock with the potential to produce 2nd generation bioethanol and bioproducts, in addition to the more conventional use for heat and/or electrical power generation. In this study, we investigated the operational parameters to maximize the production of xylo-oligosaccharides (XOS) using mild deacetylation, followed by hydrothermal pretreatment. From the laboratory to the pilot-scale, the optimized two-stage pretreatment promoted 81.5% and 70.5% hemicellulose solubilization and led to XOS yields up to 9.8% and 9.1% (w/w of initial straw), respectively. Moreover, different fungal xylanases were also tested to hydrolyze XOS into xylobiose (X2) and xylotriose (X3). GH10 from Aspergillus nidulans performed better than GH11 xylanases and the ratio of the desired products (X2 + X3) increased to 72% due to minimal monomeric sugar formation. Furthermore, a cellulose-rich fraction was obtained, which can be used in other high value-added applications, such as for the production of cello-oligomers.


Assuntos
Saccharum , Celulose , Endo-1,4-beta-Xilanases , Hidrólise , Oligossacarídeos
13.
PLoS One ; 14(2): e0212629, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30802241

RESUMO

Ferulic acid (FA), a low-molecular weight aromatic compound derived from lignin, represents a high-value molecule, used for applications in the cosmetic and pharmaceutical industries. FA can be further enzymatically converted in other commercially interesting molecules, such as vanillin and bioplastics. In several organisms, these transformations often start with a common step of FA activation via CoA-thioesterification, catalyzed by feruloyl-CoA synthetases (Fcs). In this context, these enzymes are of biotechnological interest for conversion of lignin-derived FA into high value chemicals. In this study, we describe the first structural characterization of a prokaryotic Fcs, named FCS1, isolated from a lignin-degrading microbial consortium. The FCS1 optimum pH and temperature were 9 and 37°C, respectively, with Km of 0.12 mM and Vmax of 36.82 U/mg. The circular dichroism spectra indicated a notable secondary structure stability at alkaline pH values and high temperatures. This secondary structure stability corroborates the activity data, which remains high until pH 9. The Small Angle X-Ray Scattering analyses resulted on the tertiary/quaternary structure and the low-resolution envelope in solution of FCS1, which was modeled as a homodimer using the hyperthermophilic nucleoside diphosphate-forming acetyl-CoA synthetase from Candidatus Korachaeum cryptofilum. This study contributes to the field of research by establishing the first biophysical and structural characterization for Fcs, and our data may be used for comparison against novel enzymes of this class that to be studied in the future.


Assuntos
Archaea , Proteínas Arqueais , Coenzima A Ligases , Lignina/química , Metagenoma , Microbiologia do Solo , Archaea/enzimologia , Archaea/genética , Proteínas Arqueais/química , Proteínas Arqueais/genética , Proteínas Arqueais/metabolismo , Benzaldeídos/química , Benzaldeídos/metabolismo , Coenzima A Ligases/química , Coenzima A Ligases/genética , Coenzima A Ligases/metabolismo , Ácidos Cumáricos/química , Ácidos Cumáricos/metabolismo , Concentração de Íons de Hidrogênio , Lignina/metabolismo , Domínios Proteicos , Solo
14.
Energy Sci Eng ; 7(6): 2287-2305, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32355563

RESUMO

Despite the recent discoveries of considerable fossil fuel reserves, Brazil is one of the only great economic and industrial powers with very high amounts of renewable energy in its electricity matrix. Approximately 79.3% of the electric energy supply comes from renewable resources, of which hydroelectric power represents 70.6%. The two primary concerns regarding hydroelectricity are the damage caused to the environment by the construction of dams and the uncertainty of the supply in cases of long drought seasons. This article presents an analysis on the availability and energy exploitation of sugarcane straw and forest residues derived from eucalyptus for decentralized generation using a Geographic Information System-based model. The potential bioelectricity and bioethanol production from sugarcane and eucalyptus biomass in the Administrative Region of Campinas (ARC) is higher than the demand in this region. The results provide guidelines for designing alternatives to the intended Nationally Determined Contributions in Brazil within the scope of the ARC, and they can be used to provide energy empowerment, electric matrix diversification, and new policies that address the residue availability and demand.

15.
Molecules ; 22(10)2017 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-29023400

RESUMO

A polyelectrolyte complex system of chitosan-pectin nano- and microparticles was developed to encapsulate the hormone insulin. The aim of this work was to obtain small particles for oral insulin delivery without chemical crosslinkers based on natural and biodegradable polysaccharides. The nano- and microparticles were developed using chitosans (with different degrees of acetylation: 15.0% and 28.8%) and pectin solutions at various charge ratios (n⁺/n- given by the chitosan/pectin mass ratio) and total charge. Nano- and microparticles were characterized regarding particle size, zeta potential, production yield, encapsulation efficiency, stability in different media, transmission electron microscopy and cytotoxicity assays using Caco-2 cells. The insulin release was evaluated in vitro in simulated gastric and intestinal media. Small-sized particles (~240-~1900 nm) with a maximum production yield of ~34.0% were obtained. The highest encapsulation efficiency (~62.0%) of the system was observed at a charge ratio (n⁺/n-) 5.00. The system was stable in various media, particularly in simulated gastric fluid (pH 1.2). Transmission electron microscopy (TEM) analysis showed spherical shape particles when insulin was added to the system. In simulated intestinal fluid (pH 6.8), controlled insulin release occurred over 2 h. In vitro tests indicated that the proposed system presents potential as a drug delivery for oral administration of bioactive peptides.


Assuntos
Quitosana/química , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos , Insulina/administração & dosagem , Nanopartículas/química , Pectinas/química , Eletricidade Estática , Células CACO-2 , Sobrevivência Celular/efeitos dos fármacos , Composição de Medicamentos , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Humanos , Nanopartículas/ultraestrutura , Tamanho da Partícula
17.
Genome Announc ; 4(1)2016 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-26868405

RESUMO

To shed light on the associations established between Vibrio species and soft corals in coastal ecosystems, we report here the draft genome sequence of Vibrio sp. strain Evh12, a bacterium that has been isolated from the gorgonian coral Eunicella verrucosa and that shows antagonistic activity against Escherichia coli.

18.
Carbohydr Polym ; 132: 537-45, 2015 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-26256380

RESUMO

A polyelectrolyte complex (PEC) matrix formed between chitosan and pectin was developed to entrap a bioactive compound (anthocyanin), obtaining an useful pH indicator device. Polysaccharides of opposite charges such as chitosan and pectin can have a very strong intermolecular interaction. The innovation lies in obtaining a new system based on natural and biodegradable compounds, which is simple to manufacture, to indicate variation in pH by visual changes in colour. This device has potential applications in food packaging. The PEC was studied using chitosan and pectin solutions at different pHs values (3.0, 4.0, 5.0 and 5.5) and pectin/chitosan molar ratios (1.0 to 10/1.0 to 5.0). PEC films were homogeneous and showed the highest yield (60.0%) at pH 5.5. Diffusion tests indicated efficient bioactive compound entrapment in the PEC matrix. Thermogravimetric analysis (TGA), scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy indicate the compatibility between the polymers and bioactive compound.


Assuntos
Quitosana/química , Eletrólitos/química , Pectinas/química , Concentração de Íons de Hidrogênio , Microscopia Eletrônica de Varredura , Polímeros/química , Espectroscopia de Infravermelho com Transformada de Fourier , Termogravimetria
19.
Genome Announc ; 3(3)2015 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-26044435

RESUMO

To shed light on the putative host-mediated lifestyle of the quintessential marine symbiont Aliivibrio fischeri, and on the symbiosis versus potentially pathogenic features of bacteria associated with farmed fish, we report the draft genome sequence of A. fischeri strain 5LC, a bacterium retrieved from gilthead sea bream (Sparus aurata) larvae.

20.
Genome Announc ; 3(3)2015 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-26021918

RESUMO

We report here the draft genome sequence of Vibrio sp. Vb278, a biofilm-producing strain isolated from the marine sponge Sarcotragus spinosulus, showing in vitro antibacterial activity. The annotated genome displays a range of symbiotic factors and the potential for the biosynthesis of several biologically active natural products.

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