A First Expression, Purification and Characterization of Endo-β-1,3-Glucanase from <i>Penicillium expansum</i>
β-1,3-glucanase plays an important role in the biodegradation, reconstruction, and development of β-1,3-glucan. An endo-β-1,3-glucanase which was encoded by <i>PeBgl1</i> was expressed, purified and characterized from <i>Penicillium expansum</i> for the first time. The <i&...
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| Format: | Article |
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MDPI AG
2023-09-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/9/10/961 |
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| author | Kaili Wang Siyu Huai Zhuqing Tan Guillaume Legrand Ngolong Ngea Esa Abiso Godana Jun Shi Qiya Yang Xiaoyun Zhang Lina Zhao Hongyin Zhang |
| author_facet | Kaili Wang Siyu Huai Zhuqing Tan Guillaume Legrand Ngolong Ngea Esa Abiso Godana Jun Shi Qiya Yang Xiaoyun Zhang Lina Zhao Hongyin Zhang |
| author_sort | Kaili Wang |
| collection | DOAJ |
| description | β-1,3-glucanase plays an important role in the biodegradation, reconstruction, and development of β-1,3-glucan. An endo-β-1,3-glucanase which was encoded by <i>PeBgl1</i> was expressed, purified and characterized from <i>Penicillium expansum</i> for the first time. The <i>PeBgl1</i> gene was amplified and transformed into the competent cells of <i>E. coli</i> Rosetta strain with the help of the pET-30a cloning vector. The recombinant protein PeBgl1 was expressed successfully at the induction conditions of 0.8 mmol/L IPTG at 16 °C for 16 h and then was purified by nickel ion affinity chromatography. The optimum reaction temperature of PeBgl1 was 55 °C and it had maximal activity at pH 6.0 according to the enzymatic analysis. Na<sub>2</sub>HPO<sub>4</sub>-NaH<sub>2</sub>PO<sub>4</sub> buffer (pH 6.0) and NaCl have inhibitory and enhancing effects on the enzyme activities, respectively. SDS, TritonX-100 and some metal ions (Mg<sup>2+</sup>, Ca<sup>2+</sup>, Ba<sup>2+</sup>, Cu<sup>2+</sup>, and Zn<sup>2+</sup>) have an inhibitory effect on the enzyme activity. The results showed that PeBgl1 protein has good enzyme activity at 50–60 °C and at pH 5.0–9.0, and it is not a metal dependent enzyme, which makes it robust for storage and transportation, ultimately holding great promise in green biotechnology and biorefining. |
| first_indexed | 2024-03-10T21:08:09Z |
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| institution | Directory Open Access Journal |
| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-10T21:08:09Z |
| publishDate | 2023-09-01 |
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| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj.art-ae8ce816a17e4ac1a56b6b6505ba6cb32023-11-19T17:00:16ZengMDPI AGJournal of Fungi2309-608X2023-09-0191096110.3390/jof9100961A First Expression, Purification and Characterization of Endo-β-1,3-Glucanase from <i>Penicillium expansum</i>Kaili Wang0Siyu Huai1Zhuqing Tan2Guillaume Legrand Ngolong Ngea3Esa Abiso Godana4Jun Shi5Qiya Yang6Xiaoyun Zhang7Lina Zhao8Hongyin Zhang9School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, Chinaβ-1,3-glucanase plays an important role in the biodegradation, reconstruction, and development of β-1,3-glucan. An endo-β-1,3-glucanase which was encoded by <i>PeBgl1</i> was expressed, purified and characterized from <i>Penicillium expansum</i> for the first time. The <i>PeBgl1</i> gene was amplified and transformed into the competent cells of <i>E. coli</i> Rosetta strain with the help of the pET-30a cloning vector. The recombinant protein PeBgl1 was expressed successfully at the induction conditions of 0.8 mmol/L IPTG at 16 °C for 16 h and then was purified by nickel ion affinity chromatography. The optimum reaction temperature of PeBgl1 was 55 °C and it had maximal activity at pH 6.0 according to the enzymatic analysis. Na<sub>2</sub>HPO<sub>4</sub>-NaH<sub>2</sub>PO<sub>4</sub> buffer (pH 6.0) and NaCl have inhibitory and enhancing effects on the enzyme activities, respectively. SDS, TritonX-100 and some metal ions (Mg<sup>2+</sup>, Ca<sup>2+</sup>, Ba<sup>2+</sup>, Cu<sup>2+</sup>, and Zn<sup>2+</sup>) have an inhibitory effect on the enzyme activity. The results showed that PeBgl1 protein has good enzyme activity at 50–60 °C and at pH 5.0–9.0, and it is not a metal dependent enzyme, which makes it robust for storage and transportation, ultimately holding great promise in green biotechnology and biorefining.https://www.mdpi.com/2309-608X/9/10/961endo-β-1,3-glucanase<i>Penicillium expansum</i>expression purificationenzymatic characteristics |
| spellingShingle | Kaili Wang Siyu Huai Zhuqing Tan Guillaume Legrand Ngolong Ngea Esa Abiso Godana Jun Shi Qiya Yang Xiaoyun Zhang Lina Zhao Hongyin Zhang A First Expression, Purification and Characterization of Endo-β-1,3-Glucanase from <i>Penicillium expansum</i> Journal of Fungi endo-β-1,3-glucanase <i>Penicillium expansum</i> expression purification enzymatic characteristics |
| title | A First Expression, Purification and Characterization of Endo-β-1,3-Glucanase from <i>Penicillium expansum</i> |
| title_full | A First Expression, Purification and Characterization of Endo-β-1,3-Glucanase from <i>Penicillium expansum</i> |
| title_fullStr | A First Expression, Purification and Characterization of Endo-β-1,3-Glucanase from <i>Penicillium expansum</i> |
| title_full_unstemmed | A First Expression, Purification and Characterization of Endo-β-1,3-Glucanase from <i>Penicillium expansum</i> |
| title_short | A First Expression, Purification and Characterization of Endo-β-1,3-Glucanase from <i>Penicillium expansum</i> |
| title_sort | first expression purification and characterization of endo β 1 3 glucanase from i penicillium expansum i |
| topic | endo-β-1,3-glucanase <i>Penicillium expansum</i> expression purification enzymatic characteristics |
| url | https://www.mdpi.com/2309-608X/9/10/961 |
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