Heterologous expression of an engineered truncated form of human Lewis fucosyltransferase (Fuc-TIII) by the methylotrophic yeast Pichia pastoris
A stable GS115 Pichia pastoris recombinant strain was constructed to secrete a truncated form of the human alpha(1,3/4) fucosyltransferase (amino acids 45-361). Enzyme production resulted from a secretory pathway based on the pre-pro- alpha mating factor signal sequence of the yeast Saccharomyces cerevisiae . Following its transit through the Golgi apparatus, the enzyme accumulated in the periplasmic space before its release in the culture broth (about 30 mg/l). Cell-enclosed enzyme ( approximately 0.16%) proved to be fairly stable for many freezing and thawing cycles and could be used several times as an immobilized catalyst. Soluble enzyme (>99.8%) representing the main protein of the culture broth (10%) has been characterized by Western-blotting, substrate specificities and kinetic parameters. The two forms (cell-enclosed and soluble) of recombinant enzyme may be used for in vitro synthesis of Lewisadeterminants.
| Medienart: |
Artikel |
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| Erscheinungsjahr: |
1998 |
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| Erschienen: |
1998 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:8 |
|---|---|
| Enthalten in: |
Glycobiology - 8(1998), 9 vom: 15. Sept., Seite 919-25 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Gallet, P F [VerfasserIn] |
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| Themen: |
3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase |
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| Anmerkungen: |
Date Completed 16.09.1998 Date Revised 12.05.2019 published: Print Citation Status MEDLINE |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM096148594 |
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| 100 | 1 | |a Gallet, P F |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Heterologous expression of an engineered truncated form of human Lewis fucosyltransferase (Fuc-TIII) by the methylotrophic yeast Pichia pastoris |
| 264 | 1 | |c 1998 | |
| 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 16.09.1998 | ||
| 500 | |a Date Revised 12.05.2019 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a A stable GS115 Pichia pastoris recombinant strain was constructed to secrete a truncated form of the human alpha(1,3/4) fucosyltransferase (amino acids 45-361). Enzyme production resulted from a secretory pathway based on the pre-pro- alpha mating factor signal sequence of the yeast Saccharomyces cerevisiae . Following its transit through the Golgi apparatus, the enzyme accumulated in the periplasmic space before its release in the culture broth (about 30 mg/l). Cell-enclosed enzyme ( approximately 0.16%) proved to be fairly stable for many freezing and thawing cycles and could be used several times as an immobilized catalyst. Soluble enzyme (>99.8%) representing the main protein of the culture broth (10%) has been characterized by Western-blotting, substrate specificities and kinetic parameters. The two forms (cell-enclosed and soluble) of recombinant enzyme may be used for in vitro synthesis of Lewisadeterminants | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Peptide Fragments |2 NLM | |
| 650 | 7 | |a Recombinant Proteins |2 NLM | |
| 650 | 7 | |a Fucosyltransferases |2 NLM | |
| 650 | 7 | |a EC 2.4.1.- |2 NLM | |
| 650 | 7 | |a 3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase |2 NLM | |
| 650 | 7 | |a EC 2.4.1.65 |2 NLM | |
| 700 | 1 | |a Vaujour, H |e verfasserin |4 aut | |
| 700 | 1 | |a Petit, J M |e verfasserin |4 aut | |
| 700 | 1 | |a Maftah, A |e verfasserin |4 aut | |
| 700 | 1 | |a Oulmouden, A |e verfasserin |4 aut | |
| 700 | 1 | |a Oriol, R |e verfasserin |4 aut | |
| 700 | 1 | |a Le Narvor, C |e verfasserin |4 aut | |
| 700 | 1 | |a Guilloton, M |e verfasserin |4 aut | |
| 700 | 1 | |a Julien, R |e verfasserin |4 aut | |
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