Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 2.223
Filtrar
1.
Adv Exp Med Biol ; 1345: 47-59, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582013

RESUMO

The field of tissue engineering and regenerative medicine is able to depict the mechanism of cardiac repair and development of cardiac function as well, in order to reveal findings to new therapeutic designs for clinical treatment. The foremost approach of this scientific field is the fabrication of scaffolds, which contain cells that can be used as cardiac grafts in the body, to have the preferred recovery. Cardiac tissue engineering has not been completely organized for routine clinical usages. Hence, engineering innovations hold promise to character research and treatment options in the years to come. Our group has extensive experience with regard to the structure of the heart, which makes us to our decision to continue with the preparation of heart, with the aim of developing a new ECM scaffold. Herein, we aim to assess the state-of-the-art fabrication methods, advances in decellularization and recellularization techniques. We also highlight the major achievements toward the production of a bioengineered heart obtained from decellularization and recellularization techniques.


Assuntos
Transplante de Coração , Tecidos Suporte , Bioengenharia , Matriz Extracelular , Humanos , Doadores de Tecidos , Engenharia Tecidual
2.
Nat Commun ; 12(1): 5603, 2021 09 23.
Artigo em Inglês | MEDLINE | ID: mdl-34556652

RESUMO

Although the first dissection of the human ovary dates back to the 17th century, the biophysical characteristics of the ovarian cell microenvironment are still poorly understood. However, this information is vital to deciphering cellular processes such as proliferation, morphology and differentiation, as well as pathologies like tumor progression, as demonstrated in other biological tissues. Here, we provide the first readout of human ovarian fiber morphology, interstitial and perifollicular fiber orientation, pore geometry, topography and surface roughness, and elastic and viscoelastic properties. By determining differences between healthy prepubertal, reproductive-age, and menopausal ovarian tissue, we unravel and elucidate a unique biophysical phenotype of reproductive-age tissue, bridging biophysics and female fertility. While these data enable to design of more biomimetic scaffolds for the tissue-engineered ovary, our analysis pipeline is applicable for the characterization of other organs in physiological or pathological states to reveal their biophysical markers or design their bioinspired analogs.


Assuntos
Ovário/anatomia & histologia , Ovário/fisiologia , Adulto , Fatores Etários , Idoso , Bioengenharia , Criança , Pré-Escolar , Tecido Elástico/anatomia & histologia , Tecido Elástico/metabolismo , Elasticidade , Matriz Extracelular/química , Matriz Extracelular/metabolismo , Feminino , Hormônios/metabolismo , Humanos , Pessoa de Meia-Idade , Folículo Ovariano/crescimento & desenvolvimento , Reserva Ovariana , Ovário/citologia , Viscosidade , Adulto Jovem
3.
Adv Exp Med Biol ; 1345: 35-46, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582012

RESUMO

Since lung tissue is not able to be reconstructed after substantial injury, lung transplantation often is the only alternative for treatment. Antibiotic-resistant organisms that remain in donor lungs causing infection in the immunosuppressed recipient are among the complications following transplantation. Development of strategies for whole lung regeneration is a pleasing choice particularly in patients with end-stage lung diseases. Reconstruction of lung tissue in vitro for transplantation received increased attention which could deal with the shortage of donor organs. Recent advancements in the field of tissue engineering and regenerative medicine have paved the road for beneficial alternative therapies. Our group has extensive experience with regard to the structure of the lung tissue, which makes us to our decision to continue with the preparation of lung, with the aim of developing a new ECM scaffold. Herein, we aim to review the state-of-art and the tissue engineering and regenerative medicine technology highlighting the major achievements toward the production of a bioengineered lung obtained decellularization and recellularization techniques. We have strong hopes that recent developments in the engineering of lung will lead to similar breakthroughs in the engineering of distal lung components in the future.


Assuntos
Engenharia Tecidual , Tecidos Suporte , Bioengenharia , Matriz Extracelular , Humanos , Pulmão
4.
Adv Exp Med Biol ; 1345: 61-70, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582014

RESUMO

Natural scaffold appears to have extensive functions in providing anchorage and structural requirements, as well as providing a structural support for cell adherence and cell interaction for further recellularization process. Specific methods used for decellularization process play an essential role in the efficacy of cell removal and successful preservation of ultrastructure and biomechanical properties of the tissue. Numerous scaffolding materials and fabrication techniques have been investigated for pancreatic tissue engineering. Techniques of casting, freeze drying, injection molding, and electrospinning have been also used to fabricate scaffolds. Herein, we aim to review the state-of-the-art and the tissue engineering and regenerative medicine technology highlighting the major achievements toward the production of a bioengineered pancreas obtained decellularization techniques and cell-on-scaffold technology.


Assuntos
Matriz Extracelular , Tecidos Suporte , Bioengenharia , Pâncreas , Engenharia Tecidual
5.
Nat Commun ; 12(1): 5027, 2021 08 19.
Artigo em Inglês | MEDLINE | ID: mdl-34413311

RESUMO

Engineered living materials (ELMs) based on bacterial cellulose (BC) offer a promising avenue for cheap-to-produce materials that can be programmed with genetically encoded functionalities. Here we explore how ELMs can be fabricated in a modular fashion from millimetre-scale biofilm spheroids grown from shaking cultures of Komagataeibacter rhaeticus. Here we define a reproducible protocol to produce BC spheroids with the high yield bacterial cellulose producer K. rhaeticus and demonstrate for the first time their potential for their use as building blocks to grow ELMs in 3D shapes. Using genetically engineered K. rhaeticus, we produce functionalized BC spheroids and use these to make and grow patterned BC-based ELMs that signal within a material and can sense and report on chemical inputs. We also investigate the use of BC spheroids as a method to regenerate damaged BC materials and as a way to fuse together smaller material sections of cellulose and synthetic materials into a larger piece. This work improves our understanding of BC spheroid formation and showcases their great potential for fabricating, patterning and repairing ELMs based on the promising biomaterial of bacterial cellulose.


Assuntos
Acetobacteraceae/crescimento & desenvolvimento , Bioengenharia/métodos , Biofilmes , Celulose/química , Engenharia Genética/métodos , Medicina Regenerativa/métodos , Acetobacteraceae/química , Acetobacteraceae/isolamento & purificação , Celulose/isolamento & purificação
6.
Nat Commun ; 12(1): 5140, 2021 08 26.
Artigo em Inglês | MEDLINE | ID: mdl-34446708

RESUMO

The difficulty of studying post-implantation development in mammals has sparked a flurry of activity to develop in vitro models, termed embryoids, based on self-organizing pluripotent stem cells. Previous approaches to derive embryoids either lack the physiological morphology and signaling interactions, or are unconducive to model post-gastrulation development. Here, we report a bioengineering-inspired approach aimed at addressing this gap. We employ a high-throughput cell aggregation approach to simultaneously coax mouse embryonic stem cells into hundreds of uniform epiblast-like aggregates in a solid matrix-free manner. When co-cultured with mouse trophoblast stem cell aggregates, the resulting hybrid structures initiate gastrulation-like events and undergo axial morphogenesis to yield structures, termed EpiTS embryoids, with a pronounced anterior development, including brain-like regions. We identify the presence of an epithelium in EPI aggregates as the major determinant for the axial morphogenesis and anterior development seen in EpiTS embryoids. Our results demonstrate the potential of EpiTS embryoids to study peri-gastrulation development in vitro.


Assuntos
Embrião de Mamíferos/embriologia , Camundongos/embriologia , Células-Tronco Embrionárias Murinas/citologia , Animais , Bioengenharia , Biomimética , Diferenciação Celular , Proliferação de Células , Implantação do Embrião , Embrião de Mamíferos/citologia , Corpos Embrioides/citologia , Desenvolvimento Embrionário , Feminino , Camadas Germinativas/citologia , Humanos , Morfogênese , Trofoblastos/citologia
8.
Nucleic Acids Res ; 49(15): 8987-8999, 2021 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-34358322

RESUMO

Single molecule force spectroscopy is a powerful approach to probe the structure, conformational changes, and kinetic properties of biological and synthetic macromolecules. However, common approaches to apply forces to biomolecules require expensive and cumbersome equipment and relatively large probes such as beads or cantilevers, which limits their use for many environments and makes integrating with other methods challenging. Furthermore, existing methods have key limitations such as an inability to apply compressive forces on single molecules. We report a nanoscale DNA force spectrometer (nDFS), which is based on a DNA origami hinge with tunable mechanical and dynamic properties. The angular free energy landscape of the nDFS can be engineered across a wide range through substitution of less than 5% of the strand components. We further incorporate a removable strut that enables reversible toggling of the nDFS between open and closed states to allow for actuated application of tensile and compressive forces. We demonstrate the ability to apply compressive forces by inducing a large bend in a 249bp DNA molecule, and tensile forces by inducing DNA unwrapping of a nucleosome sample. These results establish a versatile tool for force spectroscopy and robust methods for designing nanoscale mechanical devices with tunable force application.


Assuntos
DNA/química , Nanoestruturas/química , Bioengenharia , Fenômenos Biomecânicos , Nucleossomos/química , Análise Espectral
9.
Immunity ; 54(8): 1636-1651, 2021 08 10.
Artigo em Inglês | MEDLINE | ID: mdl-34348117

RESUMO

The development of effective vaccines to combat infectious diseases is a complex multi-year and multi-stakeholder process. To accelerate the development of vaccines for coronavirus disease 2019 (COVID-19), a novel pathogen emerging in late 2019 and spreading globally by early 2020, the United States government (USG) mounted an operation bridging public and private sector expertise and infrastructure. The success of the endeavor can be seen in the rapid advanced development of multiple vaccine candidates, with several demonstrating efficacy and now being administered around the globe. Here, we review the milestones enabling the USG-led effort, the methods utilized, and ensuing outcomes. We discuss the current status of COVID-19 vaccine development and provide a perspective for how partnership and preparedness can be better utilized in response to future public-health pandemic emergencies.


Assuntos
Vacinas contra COVID-19/imunologia , COVID-19/epidemiologia , COVID-19/prevenção & controle , Pesquisa , SARS-CoV-2/imunologia , Bioengenharia , Biotecnologia , Vacinas contra COVID-19/administração & dosagem , Humanos , Modelos Moleculares , Avaliação de Resultados em Cuidados de Saúde , Vigilância em Saúde Pública , Pesquisa/estatística & dados numéricos , Pesquisa/tendências , Estados Unidos/epidemiologia , Cobertura Vacinal/estatística & dados numéricos , Vacinologia
10.
Theranostics ; 11(16): 8043-8056, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34335979

RESUMO

Rationale: As a potentially life-threatening disorder, cerebral ischemia-reperfusion (I/R) injury is associated with significantly high mortality, especially the irreversible brain tissue damage associated with increased reactive oxygen radical production and excessive inflammation. Currently, the insufficiency of targeted drug delivery and "on-demand" drug release remain the greatest challenges for cerebral I/R injury therapy. Bioengineered cell membrane-based nanotherapeutics mimic and enhance natural membrane functions and represent a potentially promising approach, relying on selective interactions between receptors and chemokines and increase nanomedicine delivery efficiency into the target tissues. Methods: We employed a systematic method to synthesize biomimetic smart nanoparticles. The CXCR4-overexpressing primary mouse thoracic aorta endothelial cell (PMTAEC) membranes and RAPA@HOP were extruded through a 200 nm polycarbonate porous membrane using a mini-extruder to harvest the RAPA@BMHOP. The bioengineered CXCR4-overexpressing cell membrane-functionalized ROS-responsive nanotherapeutics, loaded with rapamycin (RAPA), were fabricated to enhance the targeted delivery to lesions with pathological overexpression of SDF-1. Results: RAPA@BMHOP exhibited a three-fold higher rate of target delivery efficacy via the CXCR4/SDF-1 axis than its non-targeting counterpart in an in vivo model. Additionally, in response to the excessive pathological ROS, nanotherapeutics could be degraded to promote "on-demand" cargo release and balance the ROS level by p-hydroxy-benzyl alcohol degradation, thereby scavenging excessive ROS and suppressing the free radical-induced focal damage and local inflammation. Also, the stealth effect of cell membrane coating functionalization on the surface resulted in extended circulation time and high stability of nanoparticles. Conclusion: The biomimetic smart nanotherapeutics with active targeting, developed in this study, significantly improved the therapeutic efficacy and biosafety profiles. Thus, these nanoparticles could be a candidate for efficient therapy of cerebral I/R injury.


Assuntos
Sistemas de Liberação de Medicamentos/métodos , Receptores CXCR4/metabolismo , Traumatismo por Reperfusão/terapia , Animais , Bioengenharia/métodos , Materiais Biomiméticos/farmacologia , Isquemia Encefálica/metabolismo , Membrana Celular/metabolismo , Liberação Controlada de Fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Nanomedicina/métodos , Nanopartículas/administração & dosagem , Espécies Reativas de Oxigênio/química , Espécies Reativas de Oxigênio/metabolismo , Receptores CXCR4/genética , Traumatismo por Reperfusão/metabolismo , Transdução de Sinais/fisiologia
11.
Int J Pharm ; 606: 120928, 2021 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-34303820

RESUMO

Biomimetic scaffolds engineering for improved collagen, epithelial cutaneous and fibrous tissue regeneration remains challenging for wound healing. To address these issues, this study aimed to report on the fabrication and characterization of electrospun of carboxymethyl guargum (CMGG), reduced graphene oxide (rGO) nanocomposite dressings loaded with curcumin for chronic wound healing applications. SEM and XRD examined the morphology of nanofibers and resulted in excellent porosity. TGA and FT-IR were done, which revealed the nanofibers' thermal and chemical interactions. CMGG, rGO nanocomposite with curcumin was investigated for in-vitro wound healing assay by scratch wound healing model using 3T3 L1 fibroblast cell lines and conducted in vitro drug-releasing studies. These nanocomposites showed 100% wound closure by the proliferation of fibroblast cell lines 3T3-L1 within 48 h and showed controlled drug release. Further, in vivo results also showed that the CMGG, rGO nanocomposite with curcumin has the potential wound healing effects. Histological studies showed that the CMGG, rGO nanocomposite with curcumin has the potential for wound healing, which indicates that the biomimetic CMGG nanofibers have an excellent healing effect on chronic wounds.


Assuntos
Curcumina , Nanocompostos , Nanofibras , Bioengenharia , Grafite , Espectroscopia de Infravermelho com Transformada de Fourier , Cicatrização
12.
Science ; 373(6550)2021 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-34210851

RESUMO

Synthetic biological networks comprising fast, reversible reactions could enable engineering of new cellular behaviors that are not possible with slower regulation. Here, we created a bistable toggle switch in Saccharomyces cerevisiae using a cross-repression topology comprising 11 protein-protein phosphorylation elements. The toggle is ultrasensitive, can be induced to switch states in seconds, and exhibits long-term bistability. Motivated by our toggle's architecture and size, we developed a computational framework to search endogenous protein pathways for other large and similar bistable networks. Our framework helped us to identify and experimentally verify five formerly unreported endogenous networks that exhibit bistability. Building synthetic protein-protein networks will enable bioengineers to design fast sensing and processing systems, allow sophisticated regulation of cellular processes, and aid discovery of endogenous networks with particular functions.


Assuntos
Bioengenharia , Mapas de Interação de Proteínas , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fosforilação , Proteínas de Saccharomyces cerevisiae/genética
15.
Sheng Wu Gong Cheng Xue Bao ; 37(7): 2571-2580, 2021 Jul 25.
Artigo em Chinês | MEDLINE | ID: mdl-34327922

RESUMO

Biochemical Reaction Engineering is a professional discipline with emphasis on both theory and practice. It is the core course in the curriculum provision for students majoring in bioengineering and plays an important role in the cultivation of professional talents. However, there are so much theory knowledge, such as formulas and deduction, that students showed poor efficiency in their study. Herein, to cater the objective command of innovative talents under new education background, and to improve the practical teaching efficiency and the students' enthusiasm as well as the practical innovation capability, the teaching team innovated and explored the teaching pattern, the teaching method as well as the teaching technologies. The teaching efficiency has been remarkably improved by introducing the virtual simulate technology, the micro-lecture, the case teaching pattern attempts and the scientific platform, which can be used as reference by other peers.


Assuntos
Currículo , Estudantes , Bioengenharia , Humanos
16.
Viruses ; 13(6)2021 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-34208387

RESUMO

Increasing antibiotic resistance in bacteria that cause zoonotic infections is a major problem for farmers rearing animals for food as well as for consumers who eat the contaminated meat resulting in food-borne infections. Bacteriophages incorporated in animal feed may help reduce carriage and infections in animals including chickens and pigs. There are, however, unmet challenges in protecting phages from processing stresses e.g., during animal feed pelleting operations and during transit of phages through the acidic gastric environment. Core-shell capsules were produced using a concentric nozzle and commercially available encapsulation equipment to fabricate capsules with phages formulated in an oil-in-water microemulsion in the core. pH-responsive capsules released the encapsulated phage cargo within 10-30 min triggered by changes in local environmental pH typically found in the lower gastrointestinal (GI) tract of animals. Acid stability of phages exposed to pH values as low as pH 1 was demonstrated. Encapsulated phages were able to withstand exposure to 95 °C wet heat thermal stress for up to 120 s, conditions typically encountered during feed pellet extrusion processing. Free phages were inactivated within 15 s under these conditions. The present study demonstrates that encapsulation of bacteriophages in core-shell pH-responsive capsules with water-in-oil emulsified phages in the core significantly improves phage viability upon exposure to processing and environmental stresses that require consideration during production of animal feed and application in animals for biocontrol. The results from this study should help guide future development of phage formulations suitable for use in animal feed for animal biocontrol applications.


Assuntos
Ração Animal , Bacteriófagos , Cápsulas , Composição de Medicamentos , Concentração de Íons de Hidrogênio , Ração Animal/análise , Ração Animal/virologia , Bioengenharia , Microbiota , Temperatura
17.
Nucleic Acids Res ; 49(13): 7765-7774, 2021 07 21.
Artigo em Inglês | MEDLINE | ID: mdl-34223901

RESUMO

In vitro molecular circuits, based on DNA-programmable chemistries, can perform an increasing range of high-level functions, such as molecular level computation, image or chemical pattern recognition and pattern generation. Most reported demonstrations, however, can only accept nucleic acids as input signals. Real-world applications of these programmable chemistries critically depend on strategies to interface them with a variety of non-DNA inputs, in particular small biologically relevant chemicals. We introduce here a general strategy to interface DNA-based circuits with non-DNA signals, based on input-translating modules. These translating modules contain a DNA response part and an allosteric protein sensing part, and use a simple design that renders them fully tunable and modular. They can be repurposed to either transmit or invert the response associated with the presence of a given input. By combining these translating-modules with robust and leak-free amplification motifs, we build sensing circuits that provide a fluorescent quantitative time-response to the concentration of their small-molecule input, with good specificity and sensitivity. The programmability of the DNA layer can be leveraged to perform DNA based signal processing operations, which we demonstrate here with logical inversion, signal modulation and a classification task on two inputs. The DNA circuits are also compatible with standard biochemical conditions, and we show the one-pot detection of an enzyme through its native metabolic activity. We anticipate that this sensitive small-molecule-to-DNA conversion strategy will play a critical role in the future applications of molecular-level circuitry.


Assuntos
Bioengenharia , DNA , Regulação Alostérica , Proteínas de Ligação a DNA , DNA Polimerase Dirigida por DNA , Exodesoxirribonucleases , Corantes Fluorescentes , Triptofano Sintase
18.
Bioengineered ; 12(1): 2836-2850, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34227905

RESUMO

Angiotensin I-converting enzyme 2 (ACE2), type II transmembrane serine protease 2 and 4 (TMPRSS2 and TMPRSS4) are important receptors for SARS-CoV-2 infection. In this study, the full-length tree shrewACE2 gene was cloned and sequenced, and its biological information was analyzed. The expression levels of ACE2, TMPRSS2 and TMPRSS4 in various tissues or organs of the tree shrew were detected. The results showed that the full-length ACE2 gene in tree shrews was 2,786 bp, and its CDS was 2,418 bp, encoding 805 amino acids. Phylogenetic analysis based on the CDS of ACE2 revealed that tree shrews were more similar to rabbits (85.93%) and humans (85.47%) but far from mice (82.81%) and rats (82.58%). In silico analysis according to the binding site of SARS-CoV-2 with the ACE2 receptor of different species predicted that tree shrews had potential SARS-CoV-2 infection possibility, which was similar to that of rabbits, cats and dogs but significantly higher than that of mice and rats. In addition, various tissues or organs of tree shrews expressed ACE2, TMPRSS2 and TMPRSS4. Among them, the kidney most highly expressed ACE2, followed by the lung and liver. The esophagus, lung, liver, intestine and kidney had relatively high expression levels of TMPRSS2 and TMPRSS4. In general, we reported for the first time the expression of ACE2, TMPRSS2 and TMPRSS4 in various tissues or organs in tree shrews. Our results revealed that tree shrews could be used as a potential animal model to study the mechanism underlying SARS-CoV-2 infection.


Assuntos
Enzima de Conversão de Angiotensina 2/genética , COVID-19/etiologia , Proteínas de Membrana/genética , SARS-CoV-2 , Serina Endopeptidases/genética , Tupaiidae/genética , Tupaiidae/metabolismo , Sequência de Aminoácidos , Enzima de Conversão de Angiotensina 2/química , Enzima de Conversão de Angiotensina 2/metabolismo , Animais , Bioengenharia , COVID-19/enzimologia , COVID-19/genética , Biologia Computacional , Modelos Animais de Doenças , Feminino , Humanos , Masculino , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , Modelos Moleculares , Filogenia , Estrutura Terciária de Proteína , Homologia de Sequência de Aminoácidos , Serina Endopeptidases/química , Serina Endopeptidases/metabolismo , Homologia Estrutural de Proteína , Distribuição Tecidual , Tupaiidae/virologia
19.
Int J Mol Sci ; 22(13)2021 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-34209892

RESUMO

Ferritin naturally exists in most organisms and can specifically recognize the transferrin 1 receptor (TfR1), which is generally highly expressed on various types of tumor cells. The pH dependent reversible assembling and disassembling property of ferritin renders it as a suitable candidate for encapsulating a variety of anticancer drugs and imaging probes. Ferritins external surface is chemically and genetically modifiable which can serve as attachment site for tumor specific targeting peptides or moieties. Moreover, the biological origin of these protein cages makes it a biocompatible nanocarrier that stabilizes and protects the enclosed particles from the external environment without provoking any toxic or immunogenic responses. Recent studies, further establish ferritin as a multifunctional nanocarrier for targeted cancer chemotherapy and phototherapy. In this review, we introduce the favorable characteristics of ferritin drug carriers, the specific targeted surface modification and a multifunctional nanocarriers combined chemotherapy with phototherapy for tumor treatment. Taken together, ferritin is a potential ideal base of engineered nanoparticles for tumor therapy and still needs to explore more on its way.


Assuntos
Antígenos CD/metabolismo , Bioengenharia/métodos , Ferritinas/metabolismo , Neoplasias/metabolismo , Receptores da Transferrina/metabolismo , Animais , Portadores de Fármacos , Composição de Medicamentos , Sistemas de Liberação de Medicamentos , Ferritinas/genética , Humanos , Concentração de Íons de Hidrogênio , Nanopartículas , Neoplasias/tratamento farmacológico
20.
Electron. j. biotechnol ; 52: 85-92, July. 2021. graf, tab
Artigo em Inglês | LILACS | ID: biblio-1283600

RESUMO

BACKGROUND: Nonribosomal peptide synthases (NRPS) can synthesize functionally diverse bioactive peptides by incorporating nonproteinogenic amino acids, offering a rich source of new drug leads. The bacterium Escherichia coli is a well-characterized production host and a promising candidate for the synthesis of nonribosomal peptides, but only limited bioprocess engineering has been reported for such molecules. We therefore developed a medium and optimized process parameters using the design of experiments (DoE) approach. RESULTS: We found that glycerol is not suitable as a carbon source for rhabdopeptide production, at least for the NRPS used for this study. Alternative carbon sources from the tricarboxylic acid cycle achieved much higher yields. DoE was used to optimize the pH and temperature in a stirred-tank reactor, revealing that optimal growth and optimal production required substantially different conditions. CONCLUSIONS: We developed a chemically defined adapted M9 medium matching the performance of complex medium (lysogeny broth) in terms of product concentration. The maximum yield in the reactor under optimized conditions was 126 mg L-1, representing a 31-fold increase compared to the first shaking-flask experiments with M9 medium and glycerol as the carbon source. Conditions that promoted cell growth tended to inhibit NRPS productivity. The challenge was therefore to find a compromise between these factors as the basis for further process development.


Assuntos
Peptídeo Sintases/metabolismo , Reatores Biológicos/microbiologia , Escherichia coli , Temperatura , Biotecnologia , Carbono/metabolismo , Modelos Estatísticos , Eletroforese em Gel de Poliacrilamida , Bioengenharia , Concentração de Íons de Hidrogênio
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...