Characterization, Semirational Design for pH Robustness, and the Application in Bioactive Peptide Production of a X-Prolyl Dipeptidyl Aminopeptidase from Lactococcus lactis MY-3
PepXLcMY-3, an X-prolyl dipeptidyl aminopeptidase derived from Lactobacillus lactis MY-3, was screened and recombinantly expressed in Escherichia coli. The enzyme could exhibit about 40% activity within the pH range of 6.0-10. To further improve the pH robustness, site E396 located in the active pocket was discovered through alanine scanning. The mutant E396I displayed both developed activity and kcat/Km. The optimal pH of E396I shifted from 6.0 to 10 compared to WT, with the relative activity within the pH range of 6.0-10 significantly increased. The site K648 was then proposed by semirational design. The activity of mutant E396I/K648D reached 4.03 U/mg. The optimal pH was restored to 6.0, and the pH stability was further improved. E396I/K648D could totally hydrolyze β-casomorphin 7 within 30 min. The hydrolysate showed 64.5% inhibition on angiotensin I converting enzyme, which was more efficient than those produced by E396I and WT, 23.2 and 44.7%, respectively.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:72 |
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| Enthalten in: |
Journal of agricultural and food chemistry - 72(2024), 13 vom: 03. Apr., Seite 7279-7290 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Gu, Shengdi [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 04.04.2024 Date Revised 04.04.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1021/acs.jafc.4c00146 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM370089839 |
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| 100 | 1 | |a Gu, Shengdi |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Characterization, Semirational Design for pH Robustness, and the Application in Bioactive Peptide Production of a X-Prolyl Dipeptidyl Aminopeptidase from Lactococcus lactis MY-3 |
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| 500 | |a Date Completed 04.04.2024 | ||
| 500 | |a Date Revised 04.04.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a PepXLcMY-3, an X-prolyl dipeptidyl aminopeptidase derived from Lactobacillus lactis MY-3, was screened and recombinantly expressed in Escherichia coli. The enzyme could exhibit about 40% activity within the pH range of 6.0-10. To further improve the pH robustness, site E396 located in the active pocket was discovered through alanine scanning. The mutant E396I displayed both developed activity and kcat/Km. The optimal pH of E396I shifted from 6.0 to 10 compared to WT, with the relative activity within the pH range of 6.0-10 significantly increased. The site K648 was then proposed by semirational design. The activity of mutant E396I/K648D reached 4.03 U/mg. The optimal pH was restored to 6.0, and the pH stability was further improved. E396I/K648D could totally hydrolyze β-casomorphin 7 within 30 min. The hydrolysate showed 64.5% inhibition on angiotensin I converting enzyme, which was more efficient than those produced by E396I and WT, 23.2 and 44.7%, respectively | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a ACE inhibitory activity | |
| 650 | 4 | |a X-prolyl-dipeptidyl aminopeptidase | |
| 650 | 4 | |a catalytic mechanism | |
| 650 | 4 | |a pH robustness | |
| 650 | 4 | |a semirational design | |
| 650 | 7 | |a Dipeptidyl-Peptidases and Tripeptidyl-Peptidases |2 NLM | |
| 650 | 7 | |a EC 3.4.14.- |2 NLM | |
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| 650 | 7 | |a Hydrolases |2 NLM | |
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| 650 | 7 | |a Aminopeptidases |2 NLM | |
| 650 | 7 | |a EC 3.4.11.- |2 NLM | |
| 700 | 1 | |a Yu, Junjie |e verfasserin |4 aut | |
| 700 | 1 | |a Du, Lei |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Daihui |e verfasserin |4 aut | |
| 700 | 1 | |a Zhao, Li |e verfasserin |4 aut | |
| 700 | 1 | |a Xie, Jingli |e verfasserin |4 aut | |
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