Purification and characterization of α-galactosidase from Aspergillus niger v. Tiegh RM48
An extracellular α-galactosidase from Aspergillus niger v. Tiegh RM48 was purified to apparent homogeneity by ammonium sulfate (80% saturation) precipitation, Phenyl Sepharose CL-4B hydrophobic column chromatography and Sephadex G-100 filtration chromatography. The specific activity of the enzyme wa...
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Zhejiang University Press
2009-03-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
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| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2009.02.005 |
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| author | XU Yao-xing LI Yan-li LIU Yong XU Shao-chun YAO Xiao-hong |
| author_facet | XU Yao-xing LI Yan-li LIU Yong XU Shao-chun YAO Xiao-hong |
| author_sort | XU Yao-xing |
| collection | DOAJ |
| description | An extracellular α-galactosidase from Aspergillus niger v. Tiegh RM48 was purified to apparent homogeneity by ammonium sulfate (80% saturation) precipitation, Phenyl Sepharose CL-4B hydrophobic column chromatography and Sephadex G-100 filtration chromatography. The specific activity of the enzyme was increased approximately 103-fold, from 26.29 IU·mg<sup>-1</sup> protein to 2722.3 IU. mg<sup>-1</sup> protein. The molecular mass of the purified enzyme as determined by SDS-PAGE was 108 kDa. The optimum pH and temperature of this α-galactosidase were pH 4.5 and 60 ℃, respectively. The enzyme was stable between pH 3.0 and pH 6.0, however, its activity was inhibited by all tested metal ions. α-Galactosidase activity was strongly inhibited by Cu<sup>2+</sup>, Fe<sup>2+</sup> and moderately inhibited by Li<sup>2+</sup>, Zn<sup>2+</sup>, Mn<sup>2+</sup>, K<sup>+</sup>, Na<sup>+</sup>, Ca<sup>2+</sup>, Mg<sup>2+</sup> and Co<sup>2+</sup>. Kinetic studies of the α-galactosidase showed that the K<sub>m</sub> and the V<sub>max</sub> for 4-nitrophenyl-α-D-galactopyranoside (pNPG) were 7.37 mmol·L<sup>-1</sup> and 5618 IU· mg<sup>-1</sup>·min<sup>-1</sup>, respectively. The N-terminal amino acids sequence of the α-galactosidase protein was determined as DEKAYSPCYY, no evidence showed the apparent sequence similarity between this enzyme and any other α-galactosidase listed in the database. |
| format | Article |
| id | doaj-art-c36dec8889ad476d9dd43c105f26126f |
| institution | Matheson Library |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2009-03-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-c36dec8889ad476d9dd43c105f26126f2025-08-01T05:35:19ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552009-03-013514715210.3785/j.issn.1008-9209.2009.02.00510089209Purification and characterization of α-galactosidase from Aspergillus niger v. Tiegh RM48XU Yao-xingLI Yan-liLIU YongXU Shao-chunYAO Xiao-hongAn extracellular α-galactosidase from Aspergillus niger v. Tiegh RM48 was purified to apparent homogeneity by ammonium sulfate (80% saturation) precipitation, Phenyl Sepharose CL-4B hydrophobic column chromatography and Sephadex G-100 filtration chromatography. The specific activity of the enzyme was increased approximately 103-fold, from 26.29 IU·mg<sup>-1</sup> protein to 2722.3 IU. mg<sup>-1</sup> protein. The molecular mass of the purified enzyme as determined by SDS-PAGE was 108 kDa. The optimum pH and temperature of this α-galactosidase were pH 4.5 and 60 ℃, respectively. The enzyme was stable between pH 3.0 and pH 6.0, however, its activity was inhibited by all tested metal ions. α-Galactosidase activity was strongly inhibited by Cu<sup>2+</sup>, Fe<sup>2+</sup> and moderately inhibited by Li<sup>2+</sup>, Zn<sup>2+</sup>, Mn<sup>2+</sup>, K<sup>+</sup>, Na<sup>+</sup>, Ca<sup>2+</sup>, Mg<sup>2+</sup> and Co<sup>2+</sup>. Kinetic studies of the α-galactosidase showed that the K<sub>m</sub> and the V<sub>max</sub> for 4-nitrophenyl-α-D-galactopyranoside (pNPG) were 7.37 mmol·L<sup>-1</sup> and 5618 IU· mg<sup>-1</sup>·min<sup>-1</sup>, respectively. The N-terminal amino acids sequence of the α-galactosidase protein was determined as DEKAYSPCYY, no evidence showed the apparent sequence similarity between this enzyme and any other α-galactosidase listed in the database.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2009.02.005<italic>Aspergillus niger</italic> v. Tiegh RM48α-galactosidasepurificationenzymatic characterization |
| spellingShingle | XU Yao-xing LI Yan-li LIU Yong XU Shao-chun YAO Xiao-hong Purification and characterization of α-galactosidase from Aspergillus niger v. Tiegh RM48 浙江大学学报. 农业与生命科学版 <italic>Aspergillus niger</italic> v. Tiegh RM48 α-galactosidase purification enzymatic characterization |
| title | Purification and characterization of α-galactosidase from Aspergillus niger v. Tiegh RM48 |
| title_full | Purification and characterization of α-galactosidase from Aspergillus niger v. Tiegh RM48 |
| title_fullStr | Purification and characterization of α-galactosidase from Aspergillus niger v. Tiegh RM48 |
| title_full_unstemmed | Purification and characterization of α-galactosidase from Aspergillus niger v. Tiegh RM48 |
| title_short | Purification and characterization of α-galactosidase from Aspergillus niger v. Tiegh RM48 |
| title_sort | purification and characterization of α galactosidase from aspergillus niger v tiegh rm48 |
| topic | <italic>Aspergillus niger</italic> v. Tiegh RM48 α-galactosidase purification enzymatic characterization |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2009.02.005 |
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