Produção, purificação, clonagem e aplicação de enzimas líticas

Fleuri, Luciana Francisco; Sato, Hélia Harumi

doi:10.1590/s0100-40422005000500026

Cited by 23 publications

(19 citation statements)

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“…Neste contexto, é importante ressaltar que métodos enzimáticos podem ser realizados em condições amenas de pressão e temperatura e para a obtenção seletiva de produtos da célula, os quais podem ser recuperados e purificados [2] . Os oligômeros formados pela ação de enzimas combinam significativas e convenientes características físico-químicas, biodegradabilidade in vivo, biocompatibilidade e principalmente ação anti-microbiana [11] .…”

Section: Determinação Da Estrutura Dos Ago's Por Espectrometria De Maunclassified

“…É um biopolímero abundante e está geralmente ligado a outros polissacarídeos e proteínas. Refere-se a uma estrutura cristalina altamente ordenada e insolúvel em água [1] .As quitinases (E.C.3.2.1.14), denominadas também de poli [1,4-(N-acetil-β-D-glicosaminide)] glicanohidrolase, catalisam a hidrólise randômica de ligações glicosídicas β-1,4 de N-acetil-β-D-glicosamina (GlcNAc) em quitinas e quitodextrinas [2] . São classificadas como endo-quitinases Resumo: O presente trabalho visou a aplicação da enzima quitinolítica purificada da linhagem Cellulosimicrobium cellulans 191 e da preparação comercial de papaína na hidrólise da quitina coloidal.…”

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Elucidação parcial da estrutura de aminoglucanooligossacarídeos (AGO's) produzidos enzimaticamente

et al. 2009

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e exo-quitinases. As endo-quitinases clivam de maneira randômica liberando oligômeros como quitotetraose, quitotriose e diacetilquitobiose. Exo-quitinases liberam progressivamente diacetilquitobiose da extremidade não redutora do polímero de quitina. Este último tipo de atividade quitinolítica foi também denominado de quitobiosidase. N-acetilglicosaminidases liberam monômeros de GlcNAc dos oligômeros resultantes da ação de endo e exoquitinases [3] .A hidrólise enzimática de polímeros por preparações comerciais e multienzimáticas para a produção de oligômeros têm sido relatadas por vários pesquisadores. A quitosana, polímero proveniente da desacetilação da quitina, foi hidro- IntroduçãoA quitina é um polímero de N-acetilglicosamina com ligações β-1,4 encontrada principalmente em carapaças de insetos, parede celular de fungos e crustáceos. É um biopolímero abundante e está geralmente ligado a outros polissacarídeos e proteínas. Refere-se a uma estrutura cristalina altamente ordenada e insolúvel em água [1] .As quitinases (E.C.3.2.1.14), denominadas também de poli [1,4-(N-acetil-β-D-glicosaminide)] glicanohidrolase, catalisam a hidrólise randômica de ligações glicosídicas β-1,4 de N-acetil-β-D-glicosamina (GlcNAc) em quitinas e quitodextrinas [2] . São classificadas como endo-quitinases Resumo: O presente trabalho visou a aplicação da enzima quitinolítica purificada da linhagem Cellulosimicrobium cellulans 191 e da preparação comercial de papaína na hidrólise da quitina coloidal. A quitina coloidal foi caracterizada quanto ao grau de desacetilação e apresentou GD de 14% por espectroscopia na região do infravermelho. A quitinase purificada hidrolisou a quitina coloidal liberando di-N-acetilquitobiose, enquanto que a preparação comercial de papaína atuando sobre o mesmo substrato formou di-N-acetilquitobiose e tri-N-acetilquitotriose. A estrutura química dos aminoglucanooligossacarídeos foi elucidada por espectrometria de massa. Palavras-chave: Quitinase, papaína, aminoglucanooligossacarídeos, quitina coloidal, espectrometria de massas. Partial Elucidation of the Chemical Structure of Aminoglucanoligosaccharides (AGO's) Produced EnzymaticallyAbstract: The aim of this work was the application of purified chitinolytic enzyme from Cellulosimicrobium cellulans 191 and the commercial papain in the hydrolysis of colloidal chitin. The latter was characterized with regard to the degree deacetylation (GD), with a GD of 14% being obtained using infrared spectroscopy. The purified chitinase hydrolysed the colloidal chitin and formed di-N-acetylchitobiose, while the commercial papain acting on the same substrate formed di-N-acetylchitobiose and tri-N-acetylchitotriose. The aminoglucanoligosaccharides chemical structure was elucidated by mass spectrometry.

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Section: Determinação Da Estrutura Dos Ago's Por Espectrometria De Maunclassified

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Elucidação parcial da estrutura de aminoglucanooligossacarídeos (AGO's) produzidos enzimaticamente

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“…and other genera of yeasts, which extends their use allowing the selective manufacture of products, regardless of scale, and can be performed under temperature and pH conditions that do not involve the denaturation of cellular products of interest (FLEURI; SATO, 2005). Mathematical models have been increasingly used to help explain responses of biochemical reactions.…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…There are numerous applications for biotechnologies: preparation of protoplasts, extraction of pigments, mass treatment of residual yeast cell fermentation industries for preparation of animal feed, in studies of the mechanism of cell wall synthesis for control of pathogenic yeast, etc. (HUNTER;ASENJO, 1988;SATO, 2005;SALAZAR; ASENJO, 2007).These enzymes are capable of lysing the cell wall of Saccharomyces cereviseae, Candida sp. and other genera of yeasts, which extends their use allowing the selective manufacture of products, regardless of scale, and can be performed under temperature and pH conditions that do not involve the denaturation of cellular products of interest (FLEURI; SATO, 2005).…”

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Application of Doehlert experimental design in the optimization of experimental variables for the Pseudozyma sp. (CCMB 306) and Pseudozyma sp. (CCMB 300) cell lysis

et al. 2012

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Application of Doehlert experimental design in the optimization of experimental variables for the Pseudozyma sp. (CCMB 306) and Pseudozyma sp. (CCMB 300) cell lysis Aplicação do modelo Doehlert na otimização das variáveis experimentais para a lise de Pseudozyma sp. (CCMB 306) e Pseudozyma sp. (CCMB 300)Amanda Reges de SENA 1,3 , Gildomar Lima VALASQUES JÚNIOR 1,2 , Ingara Keisle São Paulo BARRETTO 1 , Sandra Aparecida ASSIS 1 * IntroductionThe cell wall of yeast is basically made of glucan and mannan-protein. Lytic enzymes such as chitinases, proteases and β-1,3-glucanases are usually necessary. There are numerous applications for biotechnologies: preparation of protoplasts, extraction of pigments, mass treatment of residual yeast cell fermentation industries for preparation of animal feed, in studies of the mechanism of cell wall synthesis for control of pathogenic yeast, etc. (HUNTER;ASENJO, 1988;SATO, 2005;SALAZAR; ASENJO, 2007).These enzymes are capable of lysing the cell wall of Saccharomyces cereviseae, Candida sp. and other genera of yeasts, which extends their use allowing the selective manufacture of products, regardless of scale, and can be performed under temperature and pH conditions that do not involve the denaturation of cellular products of interest (FLEURI; SATO, 2005). Mathematical models have been increasingly used to help explain responses of biochemical reactions. This technique is often used for optimization and/or verification of the influence of medium components and cultivation parameters for enzyme production (FLEURI; SATO, 2008).The production and optimization of the parameters that affect the enzymatic synthesis should always be investigated because the optimal conditions vary among different organisms and different enzymes (BRAVO et al., 2000). In many cases, the interaction of parameters that influence fermentation processes can be evaluated with a reduced number of tests using an experimental design (THÉODORE; PANDA, 1995). Several experimental design models could be used to reduce the number of experiments under different conditions. Among them are the Plackett-Burman, Doehlert, Central Composite, and Box-Behnken (LI et al., 2007).The choice of Doehlert design is justified by a number of advantages such as (1) its spherical experimental domain with an uniformity in space filling, (2) its ability to explore the whole of the domain, and (3) its potential for sequentially where the experiments can be reused when the boundaries have not been well chosen at first (BENSALAH et al., 2010). Doehlert designs are easily applied to optimized variables and offer advantages over the Central Composite (CCD) and Box-Behnken designs, used in response surface analysis. They need fewer experiments, ResumoO presente trabalho visou verificar a influência do pH e temperatura na lise de leveduras utilizando planejamento experimental. No estudo, foi utilizado o extrato enzimático, contendo β-1,3-glucanase e quitinase líticas, obtidas do micro-organismo Moniliophthora perniciosa. O delineamento experiment...

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Extraction of carotenoids from Phaffia rhodozyma: A comparison between different techniques of cell disruption

Michelon

Borba

Rafael

et al. 2012

Food Sci Biotechnol

100

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The yeast Phaffia rhodozyma is known for producing carotenogenic pigments, commonly used in aquaculture feed formulation as well as in cosmetic, pharmaceutical, and food industries. Despite the high production of carotenoids from microorganisms by biotechnology, their use has limitation due to the cell wall resistance, which constitutes a barrier to the bioavailability of carotenoids. Therefore, there is a need to improve carotenoids recovering technique from microorganisms for the application of food industries. This study aimed to compare mechanical, chemical, and enzymatic techniques of cell disruption for extracting carotenoids produced by P. rhodozyma NRRL Y-17268. Among the techniques studied, the highest specific concentration of carotenoids (190.35 µg/g) resulted from the combined techniques of frozen biomass maceration using diatomaceous earth and enzymatic lysis at pH of the reaction medium of 4.5 at 55 o C, with initial activity of β-1,3 glucanase of 0.6 U/mL for 30 min.

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Produção, purificação, clonagem e aplicação de enzimas líticas

Cited by 23 publications

References 50 publications

Elucidação parcial da estrutura de aminoglucanooligossacarídeos (AGO's) produzidos enzimaticamente

Elucidação parcial da estrutura de aminoglucanooligossacarídeos (AGO's) produzidos enzimaticamente

Application of Doehlert experimental design in the optimization of experimental variables for the Pseudozyma sp. (CCMB 306) and Pseudozyma sp. (CCMB 300) cell lysis

Extraction of carotenoids from Phaffia rhodozyma: A comparison between different techniques of cell disruption

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