Molecular form and stability of D-hydantoinase deleted at C-terminal residue Arg
This report is about only deleting one C-terminal residue of D-hydantoinase to result in obvious changes on its molecular form and stability. A recombinant D-hydantoinase (P479) and its mutant enzyme deleted at C-terminal residue Arg (P478) were prepared by methods of gene cloning, expression and purification. Results show that the subunit molecular weight of P479 and P478 is the same (54kDa) as determined by SDS-PAGE, whilst the molecular form of native P479 and P478 is a dimer and a monomer respectively in the completely operative conditions. Compared with P479, the enzymatic activity of P478 for substrate hydantoin maintained about 40% and pH stability was obviously increased, at the alkaline side in particular, as well as the anti-SDS ability was also raised. However, the thermal stability for P478 was clearly lowed as compared to P479. It implies from above data that the C-terminal residue Arg of the D-hydantoinase is a crucial one for subunit dissociation, but non-essential for catalysis.
| Medienart: |
Artikel |
|---|
| Erscheinungsjahr: |
2006 |
|---|---|
| Erschienen: |
2006 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:46 |
|---|---|
| Enthalten in: |
Wei sheng wu xue bao = Acta microbiologica Sinica - 46(2006), 6 vom: 01. Dez., Seite 1014-7 |
| Sprache: |
Chinesisch |
|---|
| Beteiligte Personen: |
Niu, Li-xi [VerfasserIn] |
|---|
| Themen: |
368GB5141J |
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| Anmerkungen: |
Date Completed 05.05.2009 Date Revised 21.11.2013 published: Print Citation Status MEDLINE |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM168431319 |
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| 041 | |a chi | ||
| 100 | 1 | |a Niu, Li-xi |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Molecular form and stability of D-hydantoinase deleted at C-terminal residue Arg |
| 264 | 1 | |c 2006 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ohne Hilfsmittel zu benutzen |b n |2 rdamedia | ||
| 338 | |a Band |b nc |2 rdacarrier | ||
| 500 | |a Date Completed 05.05.2009 | ||
| 500 | |a Date Revised 21.11.2013 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a This report is about only deleting one C-terminal residue of D-hydantoinase to result in obvious changes on its molecular form and stability. A recombinant D-hydantoinase (P479) and its mutant enzyme deleted at C-terminal residue Arg (P478) were prepared by methods of gene cloning, expression and purification. Results show that the subunit molecular weight of P479 and P478 is the same (54kDa) as determined by SDS-PAGE, whilst the molecular form of native P479 and P478 is a dimer and a monomer respectively in the completely operative conditions. Compared with P479, the enzymatic activity of P478 for substrate hydantoin maintained about 40% and pH stability was obviously increased, at the alkaline side in particular, as well as the anti-SDS ability was also raised. However, the thermal stability for P478 was clearly lowed as compared to P479. It implies from above data that the C-terminal residue Arg of the D-hydantoinase is a crucial one for subunit dissociation, but non-essential for catalysis | ||
| 650 | 4 | |a English Abstract | |
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Recombinant Proteins |2 NLM | |
| 650 | 7 | |a Sodium Dodecyl Sulfate |2 NLM | |
| 650 | 7 | |a 368GB5141J |2 NLM | |
| 650 | 7 | |a Amidohydrolases |2 NLM | |
| 650 | 7 | |a EC 3.5.- |2 NLM | |
| 650 | 7 | |a dihydropyrimidinase |2 NLM | |
| 650 | 7 | |a EC 3.5.2.2 |2 NLM | |
| 700 | 1 | |a Zhang, Xue-yao |e verfasserin |4 aut | |
| 700 | 1 | |a Shi, Ya-wei |e verfasserin |4 aut | |
| 700 | 1 | |a Yuan, Jing-ming |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Wei sheng wu xue bao = Acta microbiologica Sinica |d 1985 |g 46(2006), 6 vom: 01. Dez., Seite 1014-7 |w (DE-627)NLM012631655 |x 0001-6209 |7 nnns |
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