Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrate
In this study, an extracellular phytase produced by Aspergillus niger 7A-1, was biochemically characterized for possible industrial application. The enzyme was purified from a crude extract obtained by solid-state fermentation (SSF) of triticale waste. The extract was obtained by microfiltration, ul...
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| Format: | Article |
| Language: | English |
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Elsevier
2018-03-01
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| Series: | Biotechnology Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215017X17301121 |
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| author | Alberto A. Neira-Vielma Cristóbal N. Aguilar Anna Ilyina Juan C. Contreras-Esquivel María das Graça Carneiro-da-Cunha Georgina Michelena-Álvarez José L. Martínez-Hernández |
| author_facet | Alberto A. Neira-Vielma Cristóbal N. Aguilar Anna Ilyina Juan C. Contreras-Esquivel María das Graça Carneiro-da-Cunha Georgina Michelena-Álvarez José L. Martínez-Hernández |
| author_sort | Alberto A. Neira-Vielma |
| collection | DOAJ |
| description | In this study, an extracellular phytase produced by Aspergillus niger 7A-1, was biochemically characterized for possible industrial application. The enzyme was purified from a crude extract obtained by solid-state fermentation (SSF) of triticale waste. The extract was obtained by microfiltration, ultrafiltration (300, 100 and 30 kDa) and DEAE-Sepharose column chromatography. The molecular weight of the purified enzyme was estimated to be 89 kDa by SDS-PAGE. The purified enzyme was most active at pH 5.3 and 56 °C, and retained 50% activity over a wide pH range of 4 to 7. The enzymatic thermostability assay showed that the enzyme retained more than 70% activity at 80 °C for 60 s, 40% activity for 120 s and 9% after 300 s. The phytase showed broad substrate specificity, a Km value of 220 μM and Vmax of 25 μM/min. The purified phytase retained 50% of its activity with phosphorylated compounds such as phenyl phosphate, 1-Naphthyl phosphate, 2-Naphthyl phosphate, p-Nitrophenyl phosphate and Glycerol-2-phosphate. The inhibition of phytase activity by metal ions was observed to be drastically inhibited (50%) by Ca++ and was slightly inhibited (10%) by Ni++, K+, and Na+, at 10 and 20 mM concentrations. A positive effect was obtained with Mg++, Mn++, Cu++, Cd++ and Ba++ at 25 and 35% with stimulatory effect on the phytase activity. |
| first_indexed | 2024-12-23T06:19:55Z |
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| institution | Directory Open Access Journal |
| issn | 2215-017X |
| language | English |
| last_indexed | 2024-12-23T06:19:55Z |
| publishDate | 2018-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biotechnology Reports |
| spelling | doaj.art-1ecb7125e89e45c48e5d41b34d7ff8a42022-12-21T17:57:12ZengElsevierBiotechnology Reports2215-017X2018-03-0117C495410.1016/j.btre.2017.12.004Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrateAlberto A. Neira-Vielma0Cristóbal N. Aguilar1Anna Ilyina2Juan C. Contreras-Esquivel3María das Graça Carneiro-da-Cunha4Georgina Michelena-Álvarez5José L. Martínez-Hernández6Group of Bioprocesses, Food Research Dept. School of Chemistry, Universidad Autónoma de Coahuila, Blvd. V. Carranza S/N. Col. República, CP 25280, Saltillo, Coahuila, México, MéxicoGroup of Bioprocesses, Food Research Dept. School of Chemistry, Universidad Autónoma de Coahuila, Blvd. V. Carranza S/N. Col. República, CP 25280, Saltillo, Coahuila, México, MéxicoCuerpo Académico de Nanobiociencias, School of Chemistry, Universidad Autónoma de Coahuila, Blvd. V. Carranza S/N. Col. República, CP 25280, Saltillo, Coahuila, MéxicoGroup of Bioprocesses, Food Research Dept. School of Chemistry, Universidad Autónoma de Coahuila, Blvd. V. Carranza S/N. Col. República, CP 25280, Saltillo, Coahuila, México, MéxicoDepartamento de Bioquímica, Universidade Federal de Pernambuco-UFPE, Av. Prof. Moraes Rego s/n, CEP 50.670-420, Recife, PE, BrazilInstituto Cubano de Investigaciones de los Derivados de la Caña de Azúcar (ICIDCA), Vía Blanca #804 y Carretera Central, Zona postal 10, código 11 000, San Miguel del Padrón Ciudad de La Habana, CubaCuerpo Académico de Nanobiociencias, School of Chemistry, Universidad Autónoma de Coahuila, Blvd. V. Carranza S/N. Col. República, CP 25280, Saltillo, Coahuila, MéxicoIn this study, an extracellular phytase produced by Aspergillus niger 7A-1, was biochemically characterized for possible industrial application. The enzyme was purified from a crude extract obtained by solid-state fermentation (SSF) of triticale waste. The extract was obtained by microfiltration, ultrafiltration (300, 100 and 30 kDa) and DEAE-Sepharose column chromatography. The molecular weight of the purified enzyme was estimated to be 89 kDa by SDS-PAGE. The purified enzyme was most active at pH 5.3 and 56 °C, and retained 50% activity over a wide pH range of 4 to 7. The enzymatic thermostability assay showed that the enzyme retained more than 70% activity at 80 °C for 60 s, 40% activity for 120 s and 9% after 300 s. The phytase showed broad substrate specificity, a Km value of 220 μM and Vmax of 25 μM/min. The purified phytase retained 50% of its activity with phosphorylated compounds such as phenyl phosphate, 1-Naphthyl phosphate, 2-Naphthyl phosphate, p-Nitrophenyl phosphate and Glycerol-2-phosphate. The inhibition of phytase activity by metal ions was observed to be drastically inhibited (50%) by Ca++ and was slightly inhibited (10%) by Ni++, K+, and Na+, at 10 and 20 mM concentrations. A positive effect was obtained with Mg++, Mn++, Cu++, Cd++ and Ba++ at 25 and 35% with stimulatory effect on the phytase activity.http://www.sciencedirect.com/science/article/pii/S2215017X17301121PhytaseBiochemical characterizationAnimal feedAspergillus nigerSolid-state fermentation |
| spellingShingle | Alberto A. Neira-Vielma Cristóbal N. Aguilar Anna Ilyina Juan C. Contreras-Esquivel María das Graça Carneiro-da-Cunha Georgina Michelena-Álvarez José L. Martínez-Hernández Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrate Biotechnology Reports Phytase Biochemical characterization Animal feed Aspergillus niger Solid-state fermentation |
| title | Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrate |
| title_full | Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrate |
| title_fullStr | Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrate |
| title_full_unstemmed | Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrate |
| title_short | Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrate |
| title_sort | purification and biochemical characterization of an aspergillus niger phytase produced by solid state fermentation using triticale residues as substrate |
| topic | Phytase Biochemical characterization Animal feed Aspergillus niger Solid-state fermentation |
| url | http://www.sciencedirect.com/science/article/pii/S2215017X17301121 |
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