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1.
Nat Commun ; 11(1): 4110, 2020 08 17.
Artigo em Inglês | MEDLINE | ID: mdl-32807790

RESUMO

Hutchinson-Gilford Progeria Syndrome (HGPS) is a premature aging disease in children that leads to early death. Smooth muscle cells (SMCs) are the most affected cells in HGPS individuals, although the reason for such vulnerability remains poorly understood. In this work, we develop a microfluidic chip formed by HGPS-SMCs generated from induced pluripotent stem cells (iPSCs), to study their vulnerability to flow shear stress. HGPS-iPSC SMCs cultured under arterial flow conditions detach from the chip after a few days of culture; this process is mediated by the upregulation of metalloprotease 13 (MMP13). Importantly, double-mutant LmnaG609G/G609GMmp13-/- mice or LmnaG609G/G609GMmp13+/+ mice treated with a MMP inhibitor show lower SMC loss in the aortic arch than controls. MMP13 upregulation appears to be mediated, at least in part, by the upregulation of glycocalyx. Our HGPS-SMCs chip represents a platform for developing treatments for HGPS individuals that may complement previous pre-clinical and clinical treatments.


Assuntos
Metaloproteinase 13 da Matriz/metabolismo , Miócitos de Músculo Liso/metabolismo , Animais , Biotecnologia/métodos , Doenças Cardiovasculares/metabolismo , Feminino , Frequência Cardíaca/efeitos dos fármacos , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/efeitos dos fármacos , Células-Tronco Pluripotentes Induzidas/metabolismo , Lamina Tipo A/genética , Lamina Tipo A/metabolismo , Masculino , Inibidores de Metaloproteinases de Matriz/farmacologia , Camundongos , Camundongos Mutantes , Miócitos de Músculo Liso/efeitos dos fármacos , Progéria/metabolismo , Progéria/patologia , Proteômica/métodos
2.
Proc Natl Acad Sci U S A ; 117(29): 16776-16781, 2020 07 21.
Artigo em Inglês | MEDLINE | ID: mdl-32636260

RESUMO

A particularly promising approach to deconstructing and fractionating lignocellulosic biomass to produce green renewable fuels and high-value chemicals pretreats the biomass with organic solvents in aqueous solution. Here, neutron scattering and molecular-dynamics simulations reveal the temperature-dependent morphological changes in poplar wood biomass during tetrahydrofuran (THF):water pretreatment and provide a mechanism by which the solvent components drive efficient biomass breakdown. Whereas lignin dissociates over a wide temperature range (>25 °C) cellulose disruption occurs only above 150 °C. Neutron scattering with contrast variation provides direct evidence for the formation of THF-rich nanoclusters (Rg ∼ 0.5 nm) on the nonpolar cellulose surfaces and on hydrophobic lignin, and equivalent water-rich nanoclusters on polar cellulose surfaces. The disassembly of the amphiphilic biomass is thus enabled through the local demixing of highly functional cosolvents, THF and water, which preferentially solvate specific biomass surfaces so as to match the local solute polarity. A multiscale description of the efficiency of THF:water pretreatment is provided: matching polarity at the atomic scale prevents lignin aggregation and disrupts cellulose, leading to improvements in deconstruction at the macroscopic scale.


Assuntos
Biotecnologia/métodos , Lignina/química , Madeira/química , Proteínas de Bactérias/metabolismo , Biomassa , Celulase/metabolismo , Furanos/química , Gluconacetobacter xylinus/enzimologia , Hidrólise , Lignina/metabolismo , Populus/química , Solventes/química , Tensoativos/química
3.
PLoS One ; 15(7): e0234964, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32614844

RESUMO

In this study, a facile, ecological and economical green method is described for the fabrication of iron (Fe), copper (Cu) and silver (Ag) nanoparticles (NPs) from the extract of Syzygium cumini leaves. The obtained metal NPs were categorized using UV/Vis, SEM, TEM, FTIR and EDX-ray spectroscopy techniques. The Fe-, Cu- and Ag-NPs were crystalline, spherical and size ranged from 40-52, 28-35 and 11-19 nm, respectively. The Ag-NPs showed excellent antimicrobial activities against methicillin- and vancomycin-resistance Staphylococcus aureus bacterial strains and Aspergillus flavus and A. parasiticus fungal species. Furthermore, the aflatoxins (AFs) production was also significantly inhibited when compared with the Fe- and Cu-NPs. In contrast, the adsorption results of NPs with aflatoxin B1 (AFB1) were observed as following order Fe->Cu->Ag-NPs. The Langmuir isotherm model well described the equilibrium data by the sorption capacity of Fe-NPs (105.3 ng mg-1), Cu-NPs (88.5 ng mg-1) and Ag-NPs (81.7 ng mg-1). The adsorption was found feasible, endothermic and follow the pseudo-second order kinetic model as revealed by the thermodynamic and kinetic studies. The present findings suggests that the green synthesis of metal NPs is a simple, sustainable, non-toxic, economical and energy-effective as compared to the others conventional approaches. In addition, synthesized metal NPs might be a promising AFs adsorbent for the detoxification of AFB1 in human and animal food/feed.


Assuntos
Aflatoxina B1/isolamento & purificação , Anti-Infecciosos/metabolismo , Biotecnologia/métodos , Cobre , Química Verde/métodos , Ferro , Nanopartículas Metálicas , Extratos Vegetais/metabolismo , Folhas de Planta/metabolismo , Prata , Desintoxicação por Sorção , Syzygium/metabolismo , Adsorção , Anti-Infecciosos/farmacologia , Aspergillus/efeitos dos fármacos , Aspergillus flavus/efeitos dos fármacos , Técnicas de Química Analítica , Cobre/administração & dosagem , Cobre/farmacologia , Resistência Microbiana a Medicamentos , Ferro/administração & dosagem , Ferro/farmacologia , Cinética , Nanopartículas Metálicas/administração & dosagem , Nanopartículas Metálicas/química , Nanopartículas Metálicas/ultraestrutura , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Oxirredução , Prata/administração & dosagem , Prata/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Termodinâmica , Vancomicina/farmacologia
4.
PLoS One ; 15(6): e0234870, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32569317

RESUMO

Mucoromycota fungi possess a versatile metabolism and can utilize various substrates for production of industrially important products, such as lipids, chitin/chitosan, polyphosphates, pigments, alcohols and organic acids. However, as far as commercialisation is concerned, establishing industrial biotechnological processes based on Mucoromycota fungi is still challenging due to the high production costs compared to the final product value. Therefore, the development of co-production concept is highly desired since more than one valuable product could be produced at the time and the process has a potentially higher viability. To develop such biotechnological strategy, we applied a high throughput approach consisting of micro-titre cultivation and FTIR spectroscopy. This approach allows single-step biochemical fingerprinting of either fungal biomass or growth media without tedious extraction of metabolites. The influence of two types of nitrogen sources and different levels of inorganic phosphorus on the co-production of lipids, chitin/chitosan and polyphosphates for nine different oleaginous Mucoromycota fungi was evaluated. FTIR analysis of biochemical composition of Mucoromycota fungi and biomass yield showed that variation in inorganic phosphorus had higher effect when inorganic nitrogen source-ammonium sulphate-was used. It was observed that: (1) Umbelopsis vinacea reached almost double biomass yield compared to other strains when yeast extract was used as nitrogen source while phosphorus limitation had little effect on the biomass yield; (2) Mucor circinelloides, Rhizopus stolonifer, Amylomyces rouxii, Absidia glauca and Lichtheimia corymbifera overproduced chitin/chitosan under the low pH caused by the limitation of inorganic phosphorus; (3) Mucor circinelloides, Amylomyces rouxii, Rhizopus stolonifer and Absidia glauca were able to store polyphosphates in addition to lipids when high concentration of inorganic phosphorus was used; (4) the biomass and lipid yield of high-value lipid producers Mortierella alpina and Mortierella hyalina were significantly increased when high concentrations of inorganic phosphorus were combined with ammonium sulphate, while the same amount of inorganic phosphorus combined with yeast extract showed negative impact on the growth and lipid accumulation. FTIR spectroscopy revealed the co-production potential of several oleaginous Mucoromycota fungi forming lipids, chitin/chitosan and polyphosphates in a single cultivation process.


Assuntos
Biotecnologia/métodos , Técnicas de Cultura de Células/métodos , Quitina/biossíntese , Fungos , Lipídeos/biossíntese , Polifosfatos/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Meios de Cultura , Fungos/crescimento & desenvolvimento , Fungos/metabolismo , Nitrogênio/metabolismo , Fósforo/metabolismo
5.
PLoS One ; 15(6): e0229968, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32497077

RESUMO

Although laccase has been recognized as a wonder molecule and green enzyme, the use of low yielding fungal strains, poor production, purification, and low enzyme kinetics have hampered its large-scale application. Thus,this study aims to select high yielding fungal strains and optimize the production, purification, and kinetics of laccase of Aspergillus sp. HB_RZ4. The results obtained indicated that Aspergillus sp. HB_RZ4 produced a significantly large amount of laccase under meso-acidophilic shaking conditions in a medium containing glucose and yeast extract. A 25 µM CuSO4 was observed to enhance the enzyme yield. The enzyme was best purified on a Sephadex G-100 column. The purified enzyme resembled laccase of A. flavus. The kinetics of the purified enzyme revealed high substrate specificity and good velocity of reaction,using ABTS as a substrate. The enzyme was observed to be stable over various pH values and temperatures. The peptide structure of the purified enzyme was found to resemble laccase of A. kawachii IFO 4308. The fungus was observed to decolorize various dyes independent of the requirement of a laccase mediator system.Aspergillus sp. HB_RZ4 was observed to be a potent natural producer of laccase, and it decolorized the dyes even in the absence of a laccase mediator system. Thus, it can be used for bioremediation of effluent that contains non-textile dyes.


Assuntos
Aspergillus/enzimologia , Biotecnologia/métodos , Corantes/metabolismo , Lacase/metabolismo , Casca de Planta/microbiologia , Biodegradação Ambiental , Corantes/isolamento & purificação , Ativação Enzimática , Inibidores Enzimáticos/farmacologia , Estabilidade Enzimática , Enzimas Imobilizadas/antagonistas & inibidores , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Concentração de Íons de Hidrogênio , Lacase/antagonistas & inibidores , Lacase/química
7.
Mol Cell ; 78(4): 614-623, 2020 05 21.
Artigo em Inglês | MEDLINE | ID: mdl-32442504

RESUMO

Synthetic biology has promised and delivered on an impressive array of applications based on genetically modified microorganisms. While novel biotechnology undoubtedly offers benefits, like all new technology, precautions should be considered during implementation to reduce the risk of both known and unknown adverse effects. To achieve containment of transgenic microorganisms, confidence to a near-scientific certainty that they cannot transfer their transgenic genes to other organisms, and that they cannot survive to propagate in unintended environments, is a priority. Here, we present an in-depth summary of biological containment systems for micro-organisms published to date, including the production of a genetic firewall through genome recoding and physical containment of microbes using auxotrophies, regulation of essential genes, and expression of toxic genes. The level of containment required to consider a transgenic organism suitable for deployment is discussed, as well as standards of practice for developing new containment systems.


Assuntos
Biotecnologia/métodos , Genes Essenciais , Engenharia Genética/métodos , Microbiota/genética , Microrganismos Geneticamente Modificados/genética , Biologia Sintética/métodos , Humanos , Microrganismos Geneticamente Modificados/crescimento & desenvolvimento
9.
Nat Commun ; 11(1): 2144, 2020 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-32358542

RESUMO

The Saccharomycotina subphylum (budding yeasts) spans 400 million years of evolution and includes species that thrive in diverse environments. To study niche-adaptation, we identify changes in gene expression in three divergent yeasts grown in the presence of various stressors. Duplicated and non-conserved genes are significantly more likely to respond to stress than genes that are conserved as single-copy orthologs. Next, we develop a sorting method that considers evolutionary origin and duplication timing to assign an evolutionary age to each gene. Subsequent analysis reveals that genes that emerged in recent evolutionary time are enriched amongst stress-responsive genes for each species. This gene expression pattern suggests that budding yeasts share a stress adaptation mechanism, whereby selective pressure leads to functionalization of young genes to improve growth in adverse conditions. Further characterization of young genes from species that thrive in harsh environments can inform the design of more robust strains for biotechnology.


Assuntos
Saccharomycetales/metabolismo , Ascomicetos/genética , Ascomicetos/metabolismo , Biotecnologia/métodos , Genoma Fúngico/genética , Filogenia , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo
10.
Nat Commun ; 11(1): 1529, 2020 03 23.
Artigo em Inglês | MEDLINE | ID: mdl-32251274

RESUMO

Inteins are protein segments capable of joining adjacent residues via a peptide bond. In this process known as protein splicing, the intein itself is not present in the final sequence, thus achieving scarless peptide ligation. Here, we assess the splicing activity of 34 inteins (both uncharacterized and known) using a rapid split fluorescent reporter characterization platform, and establish a library of 15 mutually orthogonal split inteins for in vivo applications, 10 of which can be simultaneously used in vitro. We show that orthogonal split inteins can be coupled to multiple split transcription factors to implement complex logic circuits in living organisms, and that they can also be used for the in vitro seamless assembly of large repetitive proteins with biotechnological relevance. Our work demonstrates the versatility and vast potential of an expanded library of orthogonal split inteins for their use in the fields of synthetic biology and protein engineering.


Assuntos
Biotecnologia/métodos , Biblioteca Gênica , Inteínas/genética , Engenharia de Proteínas/métodos , Processamento de Proteína , Clonagem Molecular , Estudos de Viabilidade , Fluorescência , Genes Reporter/genética , Proteínas Luminescentes/química , Proteínas Luminescentes/genética , Peptídeos , Plasmídeos/genética , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Fatores de Transcrição/genética , Transformação Bacteriana
11.
Diabetes Metab Syndr ; 14(4): 419-422, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32344370

RESUMO

BACKGROUND AND AIMS: COVID 19 (Coronavirus) pandemic has created surge demand for essential healthcare equipment, medicines along with the requirement for advance information technologies applications. Industry 4.0 is known as the fourth industrial revolution, which has the potential to fulfil customised requirement during COVID-19 crisis. This revolution has started with the applications of advance manufacturing and digital information technologies. METHODS: A detailed review of the literature is done on the technologies of Industry 4.0 and their applications in the COVID-19 pandemic, using appropriate search words on the databases of PubMed, SCOPUS, Google Scholar and Research Gate. RESULTS: We found several useful technologies of Industry 4.0 which help for proper control and management of COVID-19 pandemic and these have been discussed in this paper. The available technologies of Industry 4.0 could also help the detection and diagnosis of COVID-19 and other related problems and symptoms. CONCLUSIONS: Industry 4.0 can fulfil the requirements of customised face masks, gloves, and collect information for healthcare systems for proper controlling and treating of COVID-19 patients. We have discussed ten major technologies of Industry 4.0 which help to solve the problems of this virus. It is useful to provide day to day update of an infected patient, area-wise, age-wise and state-wise with proper surveillance systems. We also believe that the proper implementation of these technologies would help to enhance education and communication regarding public health. These Industry 4.0 technologies could provide a lot of innovative ideas and solution for fighting local and global medical emergencies.


Assuntos
Biotecnologia/métodos , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/prevenção & controle , Assistência à Saúde/normas , Controle de Infecções/organização & administração , Tecnologia da Informação/estatística & dados numéricos , Pandemias/prevenção & controle , Educação de Pacientes como Assunto , Pneumonia Viral/diagnóstico , Pneumonia Viral/prevenção & controle , Betacoronavirus/patogenicidade , Técnicas de Laboratório Clínico/normas , Infecções por Coronavirus/tratamento farmacológico , Infecções por Coronavirus/transmissão , Infecções por Coronavirus/virologia , Humanos , Controle de Infecções/tendências , Assistência ao Paciente , Pneumonia Viral/transmissão , Pneumonia Viral/virologia , Saúde Pública , Telemedicina
12.
Cell Prolif ; 53(5): e12785, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32339373

RESUMO

Regenerative medicine using human or porcine ß-cells or islets has an excellent potential to become a clinically relevant method for the treatment of type-1 diabetes. High-resolution imaging of the function and faith of transplanted porcine pancreatic islets and human stem cell-derived beta cells in large animals and patients for testing advanced therapy medicinal products (ATMPs) is a currently unmet need for pre-clinical/clinical trials. The iNanoBIT EU H2020 project is developing novel highly sensitive nanotechnology-based imaging approaches allowing for monitoring of survival, engraftment, proliferation, function and whole-body distribution of the cellular transplants in a porcine diabetes model with excellent translational potential to humans. We develop and validate the application of single-photon emission computed tomography (SPECT) and optoacoustic imaging technologies in a transgenic insulin-deficient pig model to observe transplanted porcine xeno-islets and in vitro differentiated human beta cells. We are progressing in generating new transgenic reporter pigs and human-induced pluripotent cell (iPSC) lines for optoacoustic imaging and testing them in transplantable bioartificial islet devices. Novel multifunctional nanoparticles have been generated and are being tested for nuclear imaging of islets and beta cells using a new, high-resolution SPECT imaging device. Overall, the combined multidisciplinary expertise of the project partners allows progress towards creating much needed technological toolboxes for the xenotransplantation and ATMP field, and thus reinforces the European healthcare supply chain for regenerative medicinal products.


Assuntos
Biotecnologia/métodos , Diabetes Mellitus Tipo 1/terapia , Transplante das Ilhotas Pancreáticas/métodos , Ilhotas Pancreáticas/cirurgia , Nanotecnologia/métodos , Animais , Animais Geneticamente Modificados , Humanos , Medicina Regenerativa/métodos , Suínos
13.
Phys Chem Chem Phys ; 22(8): 4574-4580, 2020 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-32048659

RESUMO

The organization of multiple subcellular compartments is controlled by liquid-liquid phase separation. Phase separation of this type occurs with the emergence of interfacial tension. Aqueous two-phase systems formed by two non-ionic polymers can be used to separate and analyze biological macromolecules, cells and viruses. Phase separation in these systems may serve as the simple model of phase separation in cells also occurring in aqueous media. To better understand liquid-liquid phase separation mechanisms, interfacial tension was measured in aqueous two-phase systems formed by dextran and polyethylene glycol and by polyethylene glycol and sodium sulfate in the presence of different additives. Interfacial tension values depend on differences between the solvent properties of the coexisting phases, estimated experimentally by parameters representing dipole-dipole, ion-dipole, ion-ion, and hydrogen bonding interactions. Based on both current and literature data, we propose a mechanism for phase separation in aqueous two-phase systems. This mechanism is based on the fundamental role of intermolecular forces. Although it remains to be confirmed, it is possible that these may underlie all liquid-liquid phase separation processes in biology.


Assuntos
Biotecnologia/métodos , Extração Líquido-Líquido , Água/química , Separação Celular , Dextranos/química , Complexos Multiproteicos/química , Complexos Multiproteicos/isolamento & purificação , Polietilenoglicóis/química , Sulfatos/química , Tensão Superficial , Vírus/isolamento & purificação
14.
Arch Microbiol ; 202(5): 1203-1209, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32077990

RESUMO

Selenium nanoparticles (Se NPs) were synthesized using Saccharomyces cerevisiae yeast. Influences of different amounts of sodium selenite (5.0, 10.0, 15.0, 20.0, and 25 µg) were evaluated on growth of yeast during incubation at 32 °C, during 4 days. UV-Vis spectroscopy results have shown that synthesized Se NPs had broad emission peak (λmax) in the wavelength around 350 nm which demonstrated that formation of Se NPs occurred in intracellular manner. Physico-chemical characteristics of the synthesized Se NPs using dynamic light scattering particle-size analyzer indicated that the fabricated Se NPs had particle size, polydispersity index, and zeta potential ranging from 75 to 709 nm, 0.189 to 0989, and -7.06 to -10.3 mV, respectively. Obtained results revealed that intracellular Se NPs with minimum particle size (75 nm), maximum zeta potential (-10.3 mV), and antioxidant activity (48.5%) were synthesized using minimum amount of selenium salt (5 µg). However, most uniform Se NPs were formed using maximum amount of selenium salt (25 µg). Results also indicated that by increasing amount of sodium selenite in the culture media, from 5.0 to 25 µg, antioxidant activity of the formed Se NPs decreased from 48.5 to 20.8, respectively.


Assuntos
Biotecnologia/métodos , Microbiologia Industrial/métodos , Nanopartículas/química , Saccharomyces cerevisiae/metabolismo , Selênio/química , Meios de Cultura , Tamanho da Partícula
15.
Mol Biotechnol ; 62(4): 219-227, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32103426

RESUMO

The ribosome is an essential organelle in charge of the translational processes in all kinds of cells. Currently, the scenario of its function has been significantly expanded from the classic machine for protein synthesis to a regulatory platform for quality control to maintain the protein homeostasis in a living cell. The ribosome is much more than a mechanical device with a static structure: it is inherently dynamic in structure and function, especially in response to the environmental fluctuations. Considerable effort has been made to regulate its structure and physiological function by engineering the components of a ribosome. The findings of the pioneering studies significantly deepened our understanding of a ribosome and exemplified how a ribosome could be engineered for biotechnology purposes in the era of synthetic biology. The engineering of ribosome offered highly accessible methods capable of comprehensively optimizing the performance of strains of industrial importance. In this article, the relevant recent advances were systematically reviewed.


Assuntos
Aminoácidos/química , Biotecnologia/métodos , Biossíntese de Proteínas , Ribossomos/química , Ribossomos/enzimologia , Biologia Sintética/métodos , Aminoácidos/síntese química , Aminoácidos/metabolismo , Códon sem Sentido/química , Códon sem Sentido/genética , Farmacorresistência Bacteriana/genética , Engenharia Metabólica/métodos , Engenharia de Proteínas/métodos , RNA Catalítico/biossíntese , RNA Catalítico/química , RNA Catalítico/genética , RNA Ribossômico/química , RNA de Transferência/química , RNA de Transferência/genética , Ribossomos/metabolismo
16.
Electron. j. biotechnol ; 43: 1-7, Jan. 2020. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1087465

RESUMO

Background: Biotechnological processes are part of modern industry as well as stricter environmental requirements. The need to reduce production costs and pollution demands for alternatives that involve the integral use of agro-industrial waste to produce bioactive compounds. The citrus industry generates large amounts of wastes due to the destruction of the fruits by microorganisms and insects together with the large amounts of orange waste generated during the production of juice and for sale fresh. The aim of this study was used orange wastes rich in polyphenolic compounds can be used as source carbon of Aspergillus fumigatus MUM 1603 to generate high added value compounds, for example, ellagic acid and other molecules of polyphenolic origin through submerged fermentation system. Results: The orange peel waste had a high concentration of polyphenols, 28% being condensed, 27% ellagitannins, 25% flavonoids and 20% gallotannins. The major polyphenolic compounds were catechin, EA and quercetin. The conditions, using an experimental design of central compounds, that allow the production of the maximum concentration of EA (18.68 mg/g) were found to be: temperature 30°C, inoculum 2 × 107 (spores/g) and orange peel polyphenols 6.2 (g/L). Conclusion: The submerged fermentation process is an effective methodology for the biotransformation of molecules present in orange waste to obtain high value-added as ellagic acid that can be used as powerful antioxidants, antibacterial and other applications.


Assuntos
Gerenciamento de Resíduos , Citrus sinensis/química , Ácido Elágico , Aspergillus fumigatus , Resíduos/análise , Flavonoides/análise , Biotecnologia/métodos , Taninos Hidrolisáveis/análise , Fermentação , Polifenóis/análise , Compostos Fitoquímicos
18.
Microb Cell Fact ; 19(1): 11, 2020 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-31964372

RESUMO

BACKGROUND: In recent years, the industrial workhorse Corynebacterium glutamicum has gained increasing interest as a host organism for the secretory production of heterologous proteins. Generally, the yield of a target protein in the culture supernatant depends on a multitude of interdependent biological and bioprocess parameters which have to be optimized. So far, the monitoring of such optimization processes depends on the availability of a direct assay for the respective target protein that can be handled also in high throughput approaches. Since simple assays, such as standard enzymatic activity assays, are not always at hand, the availability of a general protein secretion biosensor is highly desirable. RESULTS: High level secretion of proteins via the Sec protein export pathway leads to secretion stress, a phenomenon that is thought to be caused by the accumulation of incompletely or misfolded proteins at the membrane-cell envelope interface. We have analyzed the transcriptional responses of C. glutamicum to the secretory production of two different heterologous proteins and found that, in both cases, the expression of the gene encoding a homologue of the extracytosolic HtrA protease was highly upregulated. Based on this finding, a C. glutamicum Sec secretion biosensor strain was constructed in which the htrA gene on the chromosome was replaced by the eyfp gene. The fluorescence of the resulting reporter strain responded to the secretion of different heterologous proteins (cutinase from Fusarium solani pisi and alkaline phosphatase PhoA from Escherichia coli) in a dose-dependent manner. In addition, three differently efficient signal peptides for the secretory production of the cutinase could be differentiated by the biosensor signal. Furthermore, we have shown that an efficient signal peptide can be separated from a poor signal peptide by using the biosensor signal of the respective cells in fluorescence activated cell sorting experiments. CONCLUSIONS: We have succeeded in the construction of a C. glutamicum biosensor strain that allows for the monitoring of Sec-dependent secretion of heterologous proteins in a dose-dependent manner, independent of a direct assay for the desired target protein.


Assuntos
Proteínas de Bactérias/biossíntese , Técnicas Biossensoriais , Corynebacterium glutamicum/metabolismo , Proteínas Recombinantes/biossíntese , Proteínas de Bactérias/metabolismo , Biotecnologia/métodos , Engenharia Genética , Sinais Direcionadores de Proteínas , Proteínas Recombinantes/metabolismo , Via Secretória , Proteínas de Transporte Vesicular/metabolismo
19.
Braz J Med Biol Res ; 53(2): e8749, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31994600

RESUMO

For a long time, vaccines have been the main mode of defense and protection against several bacterial, viral, and parasitic diseases. However, the process of production and purification makes them expensive and unaffordable to many developing nations. An edible vaccine is when the antigen is expressed in the edible part of the plant. This reduces the cost of production of the vaccine because of ease of culturing. In this article, various types of edible vaccines that include algal and probiotics in addition to plants are discussed. Various diseases against which research has been carried out are also reviewed. This article focused on the conception of edible vaccines highlighting the various ways by which vaccines can be delivered.


Assuntos
Controle de Doenças Transmissíveis/métodos , Vacinas de Plantas Comestíveis , Biotecnologia/métodos , Humanos , Plantas Geneticamente Modificadas/genética , Vacinas de Plantas Comestíveis/administração & dosagem , Vacinas de Plantas Comestíveis/química , Vacinas de Plantas Comestíveis/imunologia
20.
Sci Rep ; 10(1): 1418, 2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-31996729

RESUMO

Coloration in insects provides a fruitful opportunity for interdisciplinary research involving both physics and biology, and for a better understanding of the design principles of biological structures. In this research we used nanometric and micrometric analyses to investigate the morphological and mechanical properties of the black-orange-black (BOB) color pattern in scelionid wasps, which has never been studied. The primary objective of the present investigation was to explore the structural and mechanical differences in the mesoscutum of four species: Baryconus with an orange mesosoma (i.e. BOB pattern), all black Baryconus, Scelio with an orange mesosoma (i.e. BOB pattern), and all black Scelio. The most outstanding findings include the absence of multilayer structures that generate structural color, a pigment concentrated in the upper surface of the epicuticle, and surprising differences between the four species. Three of the four species showed an accordion-like structure in the furrow (notaulus), whereas the adjacent mesoscutum was different in each species. Moreover, the normalized color component spectra for blue, green and red colors of the black mesoscutum of each genus showed the same spectral dependence while the orange color manifested small changes in the dominant wavelength, resulting in slightly different orange tones.


Assuntos
Biotecnologia/métodos , Nanotecnologia/métodos , Óptica e Fotônica/métodos , Vespas/química , Animais , Cor , Comunicação Interdisciplinar , Microscopia Eletrônica de Varredura , Microespectrofotometria , Vespas/ultraestrutura
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