Functional expression, purification, biochemical and biophysical characterizations, and molecular dynamics simulation of a histidine acid phosphatase from Saccharomyces cerevisiae
A histidine acid phosphatase (HAP) (PhySc) with 99.50% protein sequence similarity with PHO5 from Saccharomyces cerevisiae was expressed functionally with the molecular mass of ∼110 kDa through co-expression along with the set of molecular chaperones dnaK, dnaJ, GroESL. The purified HAP illustrated...
| Main Authors: | , , , , , , , |
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| Format: | Article |
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Springer Science and Business Media
2024
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| Online Access: | http://psasir.upm.edu.my/id/eprint/112420/ |
| _version_ | 1848865938269536256 |
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| author | Nezhad, Nima Ghahremani Jamaludin, Siti Zahra Raja Abd Rahman, Raja Noor Zaliha Mohd Yahaya, Normi Oslan, Siti Nurbaya Mohd Shariff, Fairolniza Mat Isa, Nurulfiza Leow, Thean Chor |
| author_facet | Nezhad, Nima Ghahremani Jamaludin, Siti Zahra Raja Abd Rahman, Raja Noor Zaliha Mohd Yahaya, Normi Oslan, Siti Nurbaya Mohd Shariff, Fairolniza Mat Isa, Nurulfiza Leow, Thean Chor |
| author_sort | Nezhad, Nima Ghahremani |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | A histidine acid phosphatase (HAP) (PhySc) with 99.50% protein sequence similarity with PHO5 from Saccharomyces cerevisiae was expressed functionally with the molecular mass of ∼110 kDa through co-expression along with the set of molecular chaperones dnaK, dnaJ, GroESL. The purified HAP illustrated the optimum activity of 28.75 ± 0.39 U/mg at pH 5.5 and 40 ˚C. The Km and Kcat values towards calcium phytate were 0.608 ± 0.09 mM and 650.89 ± 3.6 s− 1. The half-lives (T1/2) at 55 and 60 ˚C were 2.75 min and 55 s, respectively. The circular dichroism (CD) demonstrated that PhySc includes 30.5, 28.1, 21.3, and 20.1% of random coils, α-Helix, β-Turns, and β-Sheet, respectively. The Tm recorded by CD for PhySc was 56.5 ± 0.34˚C. The molecular docking illustrated that His59 and Asp322 act as catalytic residues in the PhySc. MD simulation showed that PhySc at 40 ˚C has higher structural stability over those of the temperatures 60 and 80 ˚C that support the thermodynamic in vitro investigations. Secondary structure content results obtained from MD simulation indicated that PhySc consists of 34.03, 33.09, 17.5, 12.31, and 3.05% of coil, helix, turn, sheet, and helix310, respectively, which is almost consistent with the experimental results. |
| first_indexed | 2025-11-15T14:12:39Z |
| format | Article |
| id | upm-112420 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-15T14:12:39Z |
| publishDate | 2024 |
| publisher | Springer Science and Business Media |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-1124202024-11-20T06:30:59Z http://psasir.upm.edu.my/id/eprint/112420/ Functional expression, purification, biochemical and biophysical characterizations, and molecular dynamics simulation of a histidine acid phosphatase from Saccharomyces cerevisiae Nezhad, Nima Ghahremani Jamaludin, Siti Zahra Raja Abd Rahman, Raja Noor Zaliha Mohd Yahaya, Normi Oslan, Siti Nurbaya Mohd Shariff, Fairolniza Mat Isa, Nurulfiza Leow, Thean Chor A histidine acid phosphatase (HAP) (PhySc) with 99.50% protein sequence similarity with PHO5 from Saccharomyces cerevisiae was expressed functionally with the molecular mass of ∼110 kDa through co-expression along with the set of molecular chaperones dnaK, dnaJ, GroESL. The purified HAP illustrated the optimum activity of 28.75 ± 0.39 U/mg at pH 5.5 and 40 ˚C. The Km and Kcat values towards calcium phytate were 0.608 ± 0.09 mM and 650.89 ± 3.6 s− 1. The half-lives (T1/2) at 55 and 60 ˚C were 2.75 min and 55 s, respectively. The circular dichroism (CD) demonstrated that PhySc includes 30.5, 28.1, 21.3, and 20.1% of random coils, α-Helix, β-Turns, and β-Sheet, respectively. The Tm recorded by CD for PhySc was 56.5 ± 0.34˚C. The molecular docking illustrated that His59 and Asp322 act as catalytic residues in the PhySc. MD simulation showed that PhySc at 40 ˚C has higher structural stability over those of the temperatures 60 and 80 ˚C that support the thermodynamic in vitro investigations. Secondary structure content results obtained from MD simulation indicated that PhySc consists of 34.03, 33.09, 17.5, 12.31, and 3.05% of coil, helix, turn, sheet, and helix310, respectively, which is almost consistent with the experimental results. Springer Science and Business Media 2024 Article PeerReviewed Nezhad, Nima Ghahremani and Jamaludin, Siti Zahra and Raja Abd Rahman, Raja Noor Zaliha and Mohd Yahaya, Normi and Oslan, Siti Nurbaya and Mohd Shariff, Fairolniza and Mat Isa, Nurulfiza and Leow, Thean Chor (2024) Functional expression, purification, biochemical and biophysical characterizations, and molecular dynamics simulation of a histidine acid phosphatase from Saccharomyces cerevisiae. World Journal of Microbiology and Biotechnology, 40 (6). art. no. 171. pp. 1-18. ISSN 0959-3993; eISSN: 1573-0972 https://link.springer.com/article/10.1007/s11274-024-03970-8 10.1007/s11274-024-03970-8 |
| spellingShingle | Nezhad, Nima Ghahremani Jamaludin, Siti Zahra Raja Abd Rahman, Raja Noor Zaliha Mohd Yahaya, Normi Oslan, Siti Nurbaya Mohd Shariff, Fairolniza Mat Isa, Nurulfiza Leow, Thean Chor Functional expression, purification, biochemical and biophysical characterizations, and molecular dynamics simulation of a histidine acid phosphatase from Saccharomyces cerevisiae |
| title | Functional expression, purification, biochemical and biophysical characterizations, and molecular dynamics simulation of a histidine acid phosphatase from Saccharomyces cerevisiae |
| title_full | Functional expression, purification, biochemical and biophysical characterizations, and molecular dynamics simulation of a histidine acid phosphatase from Saccharomyces cerevisiae |
| title_fullStr | Functional expression, purification, biochemical and biophysical characterizations, and molecular dynamics simulation of a histidine acid phosphatase from Saccharomyces cerevisiae |
| title_full_unstemmed | Functional expression, purification, biochemical and biophysical characterizations, and molecular dynamics simulation of a histidine acid phosphatase from Saccharomyces cerevisiae |
| title_short | Functional expression, purification, biochemical and biophysical characterizations, and molecular dynamics simulation of a histidine acid phosphatase from Saccharomyces cerevisiae |
| title_sort | functional expression, purification, biochemical and biophysical characterizations, and molecular dynamics simulation of a histidine acid phosphatase from saccharomyces cerevisiae |
| url | http://psasir.upm.edu.my/id/eprint/112420/ http://psasir.upm.edu.my/id/eprint/112420/ http://psasir.upm.edu.my/id/eprint/112420/ |