Production permeabilized cell mass of Kluyveromyces lactis and catalytic activity of β-galactosidase in lactose hydrolysis

Detalhes bibliográficos
Autor(a) principal: Fontes, Edimar A . F.
Data de Publicação: 2013
Outros Autores: Passos, Flávia M. L., Passos, Frederico J. V., Fontes, Paulo Rogério
Tipo de documento: Artigo
Idioma: por
Título da fonte: Revista do Instituto de Laticínios Cândido Tostes
Texto Completo: https://www.revistadoilct.com.br/rilct/article/view/64
Resumo: The production permeabilized cell mass of Kluyveromyces lactis containing the enzyme bgalactosidase and her catalytic activity in the lactose hydrolysis was accomplished. For obtaining of the permeabilized cell mass, cells were produced, growing in serum of UF sterile milk. To promote the permeabilization of the membrane, cells were treated with ethanol solution 50% to facilitate the contact between the enzyme and the substratum in the process lactose hydrolysis. The catalytic activity of the enzyme in the lactose hydrolysis was obtained by the lineal relationship between the reaction speed and dry weight in mg of permeabilized cell. The mass of 1mg of permeabilized cells was equal to 0.32 units of enzyme activity. The effect of the suspension of permeabilized cell in the initial speed of reaction was studied using different lactose concentrations (1, 40, 100 and 140 mM) and suspensions of permeabilized cells that varied from 0.1 to 3 mg/ ml.The reactions were accompanied in the spectrophotometer DU in 510nm, that it supplied a curve of the absorbance variation with the time of reaction. The inclination of the straight line presented by each curve corresponded the initial speeds for each concentration of permeabilized cell and lactose. It was used in that analysis a reagent colorimeter GOP-PAP that specifically reacted with the present glucose in the middle. For conversion of the absorvance reading in units of mM glucose, it was made a curve standard of glucose. The suspension of 1mg/ml of permeabilized cell added to the reaction mixture, it was selected by her lineal relationship with the reaction speed for minimum concentrations (1mM) and extreme (140 mM) of lactose. Values below 1mg/ml were appropriate for the lineal reaction between speed and initial concentration of enzyme.
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spelling Production permeabilized cell mass of Kluyveromyces lactis and catalytic activity of β-galactosidase in lactose hydrolysisProdução de massa de células permeabilizadas de Kluyveromyces lactis e atividade catalítica da bgalactosidaseb-galactosidase; lactose hydrolysis; Kluyveromyces lactisß-galactosidase; hidrólise de lactose; Kluyveromyces lactisThe production permeabilized cell mass of Kluyveromyces lactis containing the enzyme bgalactosidase and her catalytic activity in the lactose hydrolysis was accomplished. For obtaining of the permeabilized cell mass, cells were produced, growing in serum of UF sterile milk. To promote the permeabilization of the membrane, cells were treated with ethanol solution 50% to facilitate the contact between the enzyme and the substratum in the process lactose hydrolysis. The catalytic activity of the enzyme in the lactose hydrolysis was obtained by the lineal relationship between the reaction speed and dry weight in mg of permeabilized cell. The mass of 1mg of permeabilized cells was equal to 0.32 units of enzyme activity. The effect of the suspension of permeabilized cell in the initial speed of reaction was studied using different lactose concentrations (1, 40, 100 and 140 mM) and suspensions of permeabilized cells that varied from 0.1 to 3 mg/ ml.The reactions were accompanied in the spectrophotometer DU in 510nm, that it supplied a curve of the absorbance variation with the time of reaction. The inclination of the straight line presented by each curve corresponded the initial speeds for each concentration of permeabilized cell and lactose. It was used in that analysis a reagent colorimeter GOP-PAP that specifically reacted with the present glucose in the middle. For conversion of the absorvance reading in units of mM glucose, it was made a curve standard of glucose. The suspension of 1mg/ml of permeabilized cell added to the reaction mixture, it was selected by her lineal relationship with the reaction speed for minimum concentrations (1mM) and extreme (140 mM) of lactose. Values below 1mg/ml were appropriate for the lineal reaction between speed and initial concentration of enzyme.A produção de massa de célula permeabilizada de Kluyveromyces lactis contendo a enzimagalactosidase e sua atividade catalítica na hidrólise de lactose foi realizada. Para obtenção da massa de célula permeabilizada, células foram produzidas, crescendo em soro de leite UF estéril. Para promover a permeabilização da membrana, células foram tratadas com uma solução etanol 50% para facilitar o contato entre a enzima e o substrato no processo de hidrólise da lactose. A atividade catalítica da enzima na hidrólise de lactose foi obtida pela relação linear entre a velocidade de reação e peso seco em mg de célula permeabilizada. A massa de 1mg de células permeabilizadas equivaleu a 0,32 unidades de atividade de enzima. O efeito da suspensão de célula permeabilizada na velocidade inicial de reação foi estudado utilizando diferentes concentrações de lactose (1, 40, 100 e 140 mM) e suspensões de células permeabilizadas que variaram de 0,1 a 3 mg/ml. As reações foram acompanhadas no espectofotômetro DU em 510nm, que forneceu uma curva da variação de absorbância com o tempo de reação. A inclinação da reta apresentada por cada curva correspondeu as velocidades iniciais para cada concentração de célula permebilizada e de lactose. Foi utilizado nessa análise um reagente colorimétrico GOP-PAP que reagiu especificamente com a glicose presente no meio. Para conversão da leitura de absorvância em unidades de mM glicose, foi feita uma curva padrão de glicose. A concentração de 1mg/ml de célula permeabilizada adicionada à mistura de reação, foi selecionada pela sua relação linear com a velocidade de reação para concentrações mínimas (1 mM) e extremas (140 mM) de lactose. Valores abaixo de 1mg/ml foram apropriados para a reação linear entre velocidade e concentração inicial de enzima. ILCTFontes, Edimar A . F.Passos, Flávia M. L.Passos, Frederico J. V.Fontes, Paulo Rogério2013-12-19info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://www.revistadoilct.com.br/rilct/article/view/64Journal of Candido Tostes Dairy Institute; v. 63, n. 365 (2008); 13-18Revista do Instituto de Laticínios Cândido Tostes; v. 63, n. 365 (2008); 13-182238-64160100-3674reponame:Revista do Instituto de Laticínios Cândido Tostesinstname:Empresa de Pesquisa Agropecuária de Minas Gerais (EPAMIG)instacron:EPAMIGporhttps://www.revistadoilct.com.br/rilct/article/view/64/70Direitos autorais 2014 Revista do Instituto de Laticínios Cândido Tostesinfo:eu-repo/semantics/openAccess2013-12-19T16:24:53Zoai:oai.rilct.emnuvens.com.br:article/64Revistahttp://www.revistadoilct.com.br/ONGhttps://www.revistadoilct.com.br/rilct/oai||revistadoilct@epamig.br|| revistadoilct@oi.com.br2238-64160100-3674opendoar:2013-12-19T16:24:53Revista do Instituto de Laticínios Cândido Tostes - Empresa de Pesquisa Agropecuária de Minas Gerais (EPAMIG)false
dc.title.none.fl_str_mv Production permeabilized cell mass of Kluyveromyces lactis and catalytic activity of β-galactosidase in lactose hydrolysis
Produção de massa de células permeabilizadas de Kluyveromyces lactis e atividade catalítica da bgalactosidase
title Production permeabilized cell mass of Kluyveromyces lactis and catalytic activity of β-galactosidase in lactose hydrolysis
spellingShingle Production permeabilized cell mass of Kluyveromyces lactis and catalytic activity of β-galactosidase in lactose hydrolysis
Fontes, Edimar A . F.
b-galactosidase; lactose hydrolysis; Kluyveromyces lactis
ß-galactosidase; hidrólise de lactose; Kluyveromyces lactis
title_short Production permeabilized cell mass of Kluyveromyces lactis and catalytic activity of β-galactosidase in lactose hydrolysis
title_full Production permeabilized cell mass of Kluyveromyces lactis and catalytic activity of β-galactosidase in lactose hydrolysis
title_fullStr Production permeabilized cell mass of Kluyveromyces lactis and catalytic activity of β-galactosidase in lactose hydrolysis
title_full_unstemmed Production permeabilized cell mass of Kluyveromyces lactis and catalytic activity of β-galactosidase in lactose hydrolysis
title_sort Production permeabilized cell mass of Kluyveromyces lactis and catalytic activity of β-galactosidase in lactose hydrolysis
author Fontes, Edimar A . F.
author_facet Fontes, Edimar A . F.
Passos, Flávia M. L.
Passos, Frederico J. V.
Fontes, Paulo Rogério
author_role author
author2 Passos, Flávia M. L.
Passos, Frederico J. V.
Fontes, Paulo Rogério
author2_role author
author
author
dc.contributor.none.fl_str_mv

dc.contributor.author.fl_str_mv Fontes, Edimar A . F.
Passos, Flávia M. L.
Passos, Frederico J. V.
Fontes, Paulo Rogério
dc.subject.none.fl_str_mv

dc.subject.por.fl_str_mv b-galactosidase; lactose hydrolysis; Kluyveromyces lactis
ß-galactosidase; hidrólise de lactose; Kluyveromyces lactis
topic b-galactosidase; lactose hydrolysis; Kluyveromyces lactis
ß-galactosidase; hidrólise de lactose; Kluyveromyces lactis
description The production permeabilized cell mass of Kluyveromyces lactis containing the enzyme bgalactosidase and her catalytic activity in the lactose hydrolysis was accomplished. For obtaining of the permeabilized cell mass, cells were produced, growing in serum of UF sterile milk. To promote the permeabilization of the membrane, cells were treated with ethanol solution 50% to facilitate the contact between the enzyme and the substratum in the process lactose hydrolysis. The catalytic activity of the enzyme in the lactose hydrolysis was obtained by the lineal relationship between the reaction speed and dry weight in mg of permeabilized cell. The mass of 1mg of permeabilized cells was equal to 0.32 units of enzyme activity. The effect of the suspension of permeabilized cell in the initial speed of reaction was studied using different lactose concentrations (1, 40, 100 and 140 mM) and suspensions of permeabilized cells that varied from 0.1 to 3 mg/ ml.The reactions were accompanied in the spectrophotometer DU in 510nm, that it supplied a curve of the absorbance variation with the time of reaction. The inclination of the straight line presented by each curve corresponded the initial speeds for each concentration of permeabilized cell and lactose. It was used in that analysis a reagent colorimeter GOP-PAP that specifically reacted with the present glucose in the middle. For conversion of the absorvance reading in units of mM glucose, it was made a curve standard of glucose. The suspension of 1mg/ml of permeabilized cell added to the reaction mixture, it was selected by her lineal relationship with the reaction speed for minimum concentrations (1mM) and extreme (140 mM) of lactose. Values below 1mg/ml were appropriate for the lineal reaction between speed and initial concentration of enzyme.
publishDate 2013
dc.date.none.fl_str_mv 2013-12-19
dc.type.none.fl_str_mv


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dc.identifier.uri.fl_str_mv https://www.revistadoilct.com.br/rilct/article/view/64
url https://www.revistadoilct.com.br/rilct/article/view/64
dc.language.iso.fl_str_mv por
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dc.relation.none.fl_str_mv https://www.revistadoilct.com.br/rilct/article/view/64/70
dc.rights.driver.fl_str_mv Direitos autorais 2014 Revista do Instituto de Laticínios Cândido Tostes
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Direitos autorais 2014 Revista do Instituto de Laticínios Cândido Tostes
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv ILCT
publisher.none.fl_str_mv ILCT
dc.source.none.fl_str_mv Journal of Candido Tostes Dairy Institute; v. 63, n. 365 (2008); 13-18
Revista do Instituto de Laticínios Cândido Tostes; v. 63, n. 365 (2008); 13-18
2238-6416
0100-3674
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