Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Acta Scientiarum Biological Sciences |
Texto Completo: | https://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/61179 |
Resumo: | Starch processing industries use amylases, accounting for approximately 30% of the world’s enzyme market. Previously, an amylase-producing strain of Epicoccum nigrum was isolated from maize grains. Although E. nigrum amylase production is already reported in the literature, no published data on production optimization or characterization of the produced enzyme exists. The objectives of this work were to improve the amylase production by the E. nigrum PG 16 strain and to purify and characterize the produced enzyme. The E. nigrum PG 16 amylase production best conditions in submerged culture were: inoculum of 4% (v v-1) of a five-days-old stationary culture homogenate, agitation at 100 rpm, 25°C, natural light, 72 hours of incubation, starch as the carbon source, and an initial medium pH of 7.0. A molecular exclusion chromatography profile has shown the production of only one amylase, which was partially purified with ammonium precipitation and dialysis. The enzyme optima pH and temperature are 6.0 and 50°C, respectively. The partially purified enzyme lost its activity when incubated for 30 min in temperatures above 40°C, presenting a T50 of 46.25°C. The KM and Vmax of the partially purified enzyme are 1.72 mg mL-1 of starch and 0.15 mg min-1 of degraded starch, respectively. The ion Ca2+ slightly activated the studied enzyme. The ions Cu2+, Zn2+, and Fe3+ and the detergents SDS and Tween 80 acted as inhibitors of the studied enzyme. The partially purified enzyme released glucose from p-nitrophenyl α-D-glucopyranoside (p-NPG). Glucose was the enzyme’s main product from starch hydrolysis, as evidenced by thin-layer chromatography. The E. nigrum PG 16 studied enzyme is a glucoamylase and represents an alternative for enzymatic starch hydrolysis. |
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Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum starch; glucoamylase; Epicoccum nigrum; characterization.starch; glucoamylase; Epicoccum nigrum; characterization.Starch processing industries use amylases, accounting for approximately 30% of the world’s enzyme market. Previously, an amylase-producing strain of Epicoccum nigrum was isolated from maize grains. Although E. nigrum amylase production is already reported in the literature, no published data on production optimization or characterization of the produced enzyme exists. The objectives of this work were to improve the amylase production by the E. nigrum PG 16 strain and to purify and characterize the produced enzyme. The E. nigrum PG 16 amylase production best conditions in submerged culture were: inoculum of 4% (v v-1) of a five-days-old stationary culture homogenate, agitation at 100 rpm, 25°C, natural light, 72 hours of incubation, starch as the carbon source, and an initial medium pH of 7.0. A molecular exclusion chromatography profile has shown the production of only one amylase, which was partially purified with ammonium precipitation and dialysis. The enzyme optima pH and temperature are 6.0 and 50°C, respectively. The partially purified enzyme lost its activity when incubated for 30 min in temperatures above 40°C, presenting a T50 of 46.25°C. The KM and Vmax of the partially purified enzyme are 1.72 mg mL-1 of starch and 0.15 mg min-1 of degraded starch, respectively. The ion Ca2+ slightly activated the studied enzyme. The ions Cu2+, Zn2+, and Fe3+ and the detergents SDS and Tween 80 acted as inhibitors of the studied enzyme. The partially purified enzyme released glucose from p-nitrophenyl α-D-glucopyranoside (p-NPG). Glucose was the enzyme’s main product from starch hydrolysis, as evidenced by thin-layer chromatography. The E. nigrum PG 16 studied enzyme is a glucoamylase and represents an alternative for enzymatic starch hydrolysis.Starch processing industries use amylases, accounting for approximately 30% of the world’s enzyme market. Previously, an amylase-producing strain of Epicoccum nigrum was isolated from maize grains. Although E. nigrum amylase production is already reported in the literature, no published data on production optimization or characterization of the produced enzyme exists. The objectives of this work were to improve the amylase production by the E. nigrum PG 16 strain and to purify and characterize the produced enzyme. The E. nigrum PG 16 amylase production best conditions in submerged culture were: inoculum of 4% (v v-1) of a five-days-old stationary culture homogenate, agitation at 100 rpm, 25°C, natural light, 72 hours of incubation, starch as the carbon source, and an initial medium pH of 7.0. A molecular exclusion chromatography profile has shown the production of only one amylase, which was partially purified with ammonium precipitation and dialysis. The enzyme optima pH and temperature are 6.0 and 50°C, respectively. The partially purified enzyme lost its activity when incubated for 30 min in temperatures above 40°C, presenting a T50 of 46.25°C. The KM and Vmax of the partially purified enzyme are 1.72 mg mL-1 of starch and 0.15 mg min-1 of degraded starch, respectively. The ion Ca2+ slightly activated the studied enzyme. The ions Cu2+, Zn2+, and Fe3+ and the detergents SDS and Tween 80 acted as inhibitors of the studied enzyme. The partially purified enzyme released glucose from p-nitrophenyl α-D-glucopyranoside (p-NPG). Glucose was the enzyme’s main product from starch hydrolysis, as evidenced by thin-layer chromatography. The E. nigrum PG 16 studied enzyme is a glucoamylase and represents an alternative for enzymatic starch hydrolysis.Universidade Estadual De Maringá2023-03-22info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/6117910.4025/actascibiolsci.v45i1.61179Acta Scientiarum. Biological Sciences; Vol 45 (2023): Publicação contínua; e61179Acta Scientiarum. Biological Sciences; v. 45 (2023): Publicação contínua; e611791807-863X1679-9283reponame:Acta Scientiarum Biological Sciencesinstname:Universidade Estadual de Maringá (UEM)instacron:UEMenghttps://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/61179/751375155621Copyright (c) 2023 Acta Scientiarum. Biological Scienceshttp://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessMartim, Damaris Batistão Santos, Fabiane Cristina dosBarbosa-Tessmann, Ione Parra 2023-05-25T13:47:16Zoai:periodicos.uem.br/ojs:article/61179Revistahttps://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/PUBhttps://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/oai||actabiol@uem.br1807-863X1679-9283opendoar:2023-05-25T13:47:16Acta Scientiarum Biological Sciences - Universidade Estadual de Maringá (UEM)false |
dc.title.none.fl_str_mv |
Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum |
title |
Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum |
spellingShingle |
Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum Martim, Damaris Batistão starch; glucoamylase; Epicoccum nigrum; characterization. starch; glucoamylase; Epicoccum nigrum; characterization. |
title_short |
Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum |
title_full |
Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum |
title_fullStr |
Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum |
title_full_unstemmed |
Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum |
title_sort |
Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum |
author |
Martim, Damaris Batistão |
author_facet |
Martim, Damaris Batistão Santos, Fabiane Cristina dos Barbosa-Tessmann, Ione Parra |
author_role |
author |
author2 |
Santos, Fabiane Cristina dos Barbosa-Tessmann, Ione Parra |
author2_role |
author author |
dc.contributor.author.fl_str_mv |
Martim, Damaris Batistão Santos, Fabiane Cristina dos Barbosa-Tessmann, Ione Parra |
dc.subject.por.fl_str_mv |
starch; glucoamylase; Epicoccum nigrum; characterization. starch; glucoamylase; Epicoccum nigrum; characterization. |
topic |
starch; glucoamylase; Epicoccum nigrum; characterization. starch; glucoamylase; Epicoccum nigrum; characterization. |
description |
Starch processing industries use amylases, accounting for approximately 30% of the world’s enzyme market. Previously, an amylase-producing strain of Epicoccum nigrum was isolated from maize grains. Although E. nigrum amylase production is already reported in the literature, no published data on production optimization or characterization of the produced enzyme exists. The objectives of this work were to improve the amylase production by the E. nigrum PG 16 strain and to purify and characterize the produced enzyme. The E. nigrum PG 16 amylase production best conditions in submerged culture were: inoculum of 4% (v v-1) of a five-days-old stationary culture homogenate, agitation at 100 rpm, 25°C, natural light, 72 hours of incubation, starch as the carbon source, and an initial medium pH of 7.0. A molecular exclusion chromatography profile has shown the production of only one amylase, which was partially purified with ammonium precipitation and dialysis. The enzyme optima pH and temperature are 6.0 and 50°C, respectively. The partially purified enzyme lost its activity when incubated for 30 min in temperatures above 40°C, presenting a T50 of 46.25°C. The KM and Vmax of the partially purified enzyme are 1.72 mg mL-1 of starch and 0.15 mg min-1 of degraded starch, respectively. The ion Ca2+ slightly activated the studied enzyme. The ions Cu2+, Zn2+, and Fe3+ and the detergents SDS and Tween 80 acted as inhibitors of the studied enzyme. The partially purified enzyme released glucose from p-nitrophenyl α-D-glucopyranoside (p-NPG). Glucose was the enzyme’s main product from starch hydrolysis, as evidenced by thin-layer chromatography. The E. nigrum PG 16 studied enzyme is a glucoamylase and represents an alternative for enzymatic starch hydrolysis. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-03-22 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/61179 10.4025/actascibiolsci.v45i1.61179 |
url |
https://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/61179 |
identifier_str_mv |
10.4025/actascibiolsci.v45i1.61179 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/61179/751375155621 |
dc.rights.driver.fl_str_mv |
Copyright (c) 2023 Acta Scientiarum. Biological Sciences http://creativecommons.org/licenses/by/4.0 info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2023 Acta Scientiarum. Biological Sciences http://creativecommons.org/licenses/by/4.0 |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Estadual De Maringá |
publisher.none.fl_str_mv |
Universidade Estadual De Maringá |
dc.source.none.fl_str_mv |
Acta Scientiarum. Biological Sciences; Vol 45 (2023): Publicação contínua; e61179 Acta Scientiarum. Biological Sciences; v. 45 (2023): Publicação contínua; e61179 1807-863X 1679-9283 reponame:Acta Scientiarum Biological Sciences instname:Universidade Estadual de Maringá (UEM) instacron:UEM |
instname_str |
Universidade Estadual de Maringá (UEM) |
instacron_str |
UEM |
institution |
UEM |
reponame_str |
Acta Scientiarum Biological Sciences |
collection |
Acta Scientiarum Biological Sciences |
repository.name.fl_str_mv |
Acta Scientiarum Biological Sciences - Universidade Estadual de Maringá (UEM) |
repository.mail.fl_str_mv |
||actabiol@uem.br |
_version_ |
1799317390366867456 |