Encontrados 132 documentos, a visualizar página 1 de 14
Cloning expression and characterization of a chitin-binding domain
Carvalho, Vera; Gama, F. M.; Domingues, LucíliaChitosan and chitin represent a family of biopolymers, made up of β(1 → 4)-linked N-acetyl-d-glucosamine and d-glucosamine subunits. Chitin can be found in exoskeletons of insects, fungal cell walls and shells of crustaceans and is one of the most abundant natural polysaccharides in nature, second only to cellulose. On the other hand, chitosan is mainly obtained industrially by alkaline hydrolysis of chitin (Ts...
Recombinant expression and purification of the antimicrobial peptide Magainin-2
Ramos, Reinaldo Rodrigues; Moreira, Susana; Rodrigues, Ana; Gama, F. M.; Domingues, LucíliaMagainin-2 (MAG2) is a polycationic antimicrobial peptide isolated from the skin of the African clawed frog Xenopus laevis. It has a wide spectrum of antimicrobial activities against Gram-positive and Gram-negative bacteria, fungi and induces osmotic lysis of protozoa. MAG2 also possesses antiviral and antitumoral properties. These activities make this peptide a promising candidate for therapeutic applications....
The novel Fh8 and H fusion partners for soluble protein expression in Escherich...
Costa, Sofia Judite; Almeida, André; Castro, António G.; Domingues, Lucília; Besir, HüseyinThe Escherichia coli host system is an advantageous choice for simple and inexpensive recombinant protein production but it still presents bottlenecks at expressing soluble proteins from other organisms. Several efforts have been taken to overcome E. coli limitations, including the use of fusion partners that improve protein expression and solubility. New fusion technologies are emerging to complement the tradi...
Influence of trace elements supplementation on the production of recombinant fr...
Wanderley, Marcela Silvestre Outtes; Oliveira, Carla Cristina Marques de; Bruneska, Danyelly; Domingues, Lucília; Lima-Filho, José. L.; Teixeira, J. A.Frutalin, a galactose-specific lectin used to detect specific tumour markers, is a protein with low expression level in breadfruit. In the present study, fed-batch fermentation in a stirred tank bioreactor was used as a strategy to enhance protein production by a recombinant Pichia pastoris KM71H. By using this process, the production of recombinant frutalin was 4-fold higher than the value obtained in shaker f...
Reconstruction of a genome-scale metabolic model for the filamentous fungus Ash...
Gomes, Daniel G.; Dias, O.; Ferreira, E. C.; Domingues, Lucília; Rocha, I.Metabolic models are an important tool for in silico simulation of cells’ behavior. Until now, a considerable number of metabolic models were released for several microorganisms. Ashbya gossypii is an industrially relevant fungus intensively used for riboflavin production with no model reported until now. Despite the high similarity with the Saccharomyces cerevisiae genome, A. gossypii contains only 4726 protei...
Random and direct mutagenesis to enhance protein secretion in Ashbya gossypii
Ribeiro, Orquídea; Magalhães, Frederico; Aguiar, Tatiana Quinta; Wiebe, Marilyn G.; Penttilä, Merja; Domingues, LucíliaTo improve the general secretion ability of the biotechnologically relevant fungus Ashbya gossypii, random mutagenesis with ethyl methane sulfonate (EMS) was performed. The selection and screening strategy followed revealed mutants with improved secretion of heterologous Trichoderma reesei endoglucanase I (EGI), native α-amylase and/or native β-glucosidase. One mutant, S436, presented 1.4- to 2-fold increases i...
Characterization of the Ashbya gossypii secreted N-glycome and genomic insights...
Aguiar, Tatiana Quinta; Maaheimo, Hannu; Heiskanen, Annamari; Wiebe, Marilyn G.; Penttilä, Merja; Domingues, LucíliaThe riboflavin producer Ashbya gossypii is a filamentous hemiascomycete, closely related to the yeast Saccharomyces cerevisiae, that has been used as a model organism to study fungal developmental biology. It has also been explored as a host for the expression of recombinant proteins. However, although N-glycosylation plays important roles in protein secretion, morphogenesis, and the development of multicellula...
Fractionation of eucalyptus globulus wood by glycerol-water pretreatment : opti...
Romaní, Aloia; Ruíz, Héctor A.; Pereira, Francisco B.; Domingues, Lucília; Teixeira, J. A.A glycerol-organosolv process can be a good alternative for Eucalyptus wood fractionation into its main compounds, improving the enzymatic saccharification of the cellulose. A study of process variables - glycerol−water percent content, temperature, and process time - was carried out using a Box-Behnken experimental design. The cellulose obtained from pretreated solids was recovered almost quantitatively, leadi...
The Fh8 tag : a fusion partner for simple and cost-effective protein purificati...
Costa, Sofia Judite; Coelho, Eduardo; Franco, Lara Oliveira; Almeida, André; Castro, António G.; Domingues, LucíliaDownstream processing is still a major bottleneck in recombinant protein production representing most of its costs. Hence, there is a continuing demand of novel and cost-effective purification processes aiming at the recovery of pure and active target protein. In this work, a novel purification methodology is presented, using the Fh8 solubility enhancer tag as fusion handle. The binding properties of Fh8 tag to...
A novel adjuvant-free H fusion system for the production of recombinant immunog...
Costa, Sofia Judite; Silva, P.; Almeida, André; Conceição, A.; Domingues, Lucília; Castro, António G.The production of recombinant antigens in Escherichia coli and specific polyclonal antibodies for diagnosis and therapy is still a challenge for world-wide researchers. Several different strategies have been explored to improve both antigen and antibody production, all of them depending on a successful expression and immunogenicity of the antigen. Gene fusion technology attempted to address these challenges: fu...
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