Details
| Original language | English |
|---|---|
| Pages (from-to) | 1423-1430 |
| Number of pages | 8 |
| Journal | ChemBioChem |
| Volume | 6 |
| Issue number | 8 |
| Publication status | Published - 5 Aug 2005 |
Abstract
A flexible enzyme module system is presented that allows preparative access to important dTDP-activated deoxyhexoses from dTMP and sucrose. The strategic combination of the recombinant enzymes dTMP-kinase and sucrose synthase (SuSy), and the enzymes RmlB (4,6-dehydratase), RmlC (3,5-epimerase) and RmlD (4-ketoreductase) from the biosynthetic pathway of dTDP-β-L-rhamnose was optimized. The SuSy module (dTMP-kinase, SuSy, ± RmlB) yielded the precursor dTDP-α-D-glucose (2) or the biosynthetic intermediate dTDP-6-deoxy-4-keto-α-D-glucose (3) on a 0.2-0.6 g scale with overall yields of 62% and 72%, respectively. A two-step strategy in which the SuSy module was followed by the deoxysugar module (RmlC and RmlD) resulted in the synthesis of dTDP-β-L-rhamnose (4; 24.1 μmol, overall yield: 35.9%). Substitution of RmlC by DnmU from the dTDP-β-L-daunosamine pathway of Streptomyces peucetius in this module demonstrated that DnmU acts in vitro as a 3,5-epimerase with 3 as substrate to yield 4 (32.2 μmol, overall yield: 44.7%). Chemical reduction of 3 with NaBH4 gave a mixture of the C-4 epimers dTDP-α-D-quinovose (6) and dTDP-α-D-fucose (7) in a ratio of 2:1. In summary, the modular character of the presented enzyme system provides valuable compounds for the biochemical characterization of deoxysugar pathways playing a major role in microbial producers of antibiotic and antitumour agents.
Keywords
- Antibiotics, Biocatalysis, Carbohydrates, Enzymes, Glycoconjugates
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Medicine
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Chemistry(all)
- Organic Chemistry
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In: ChemBioChem, Vol. 6, No. 8, 05.08.2005, p. 1423-1430.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - An enzyme module system for the synthesis of dTDP-activated deoxysugars from dTMP and sucrose
AU - Elling, Lothar
AU - Rupprath, Carsten
AU - Günther, Nicole
AU - Römer, Ulrike
AU - Verseck, Stefan
AU - Weingarten, Petra
AU - Dräger, Gerald
AU - Kirschning, Andreas
AU - Piepersberg, Wolfgang
PY - 2005/8/5
Y1 - 2005/8/5
N2 - A flexible enzyme module system is presented that allows preparative access to important dTDP-activated deoxyhexoses from dTMP and sucrose. The strategic combination of the recombinant enzymes dTMP-kinase and sucrose synthase (SuSy), and the enzymes RmlB (4,6-dehydratase), RmlC (3,5-epimerase) and RmlD (4-ketoreductase) from the biosynthetic pathway of dTDP-β-L-rhamnose was optimized. The SuSy module (dTMP-kinase, SuSy, ± RmlB) yielded the precursor dTDP-α-D-glucose (2) or the biosynthetic intermediate dTDP-6-deoxy-4-keto-α-D-glucose (3) on a 0.2-0.6 g scale with overall yields of 62% and 72%, respectively. A two-step strategy in which the SuSy module was followed by the deoxysugar module (RmlC and RmlD) resulted in the synthesis of dTDP-β-L-rhamnose (4; 24.1 μmol, overall yield: 35.9%). Substitution of RmlC by DnmU from the dTDP-β-L-daunosamine pathway of Streptomyces peucetius in this module demonstrated that DnmU acts in vitro as a 3,5-epimerase with 3 as substrate to yield 4 (32.2 μmol, overall yield: 44.7%). Chemical reduction of 3 with NaBH4 gave a mixture of the C-4 epimers dTDP-α-D-quinovose (6) and dTDP-α-D-fucose (7) in a ratio of 2:1. In summary, the modular character of the presented enzyme system provides valuable compounds for the biochemical characterization of deoxysugar pathways playing a major role in microbial producers of antibiotic and antitumour agents.
AB - A flexible enzyme module system is presented that allows preparative access to important dTDP-activated deoxyhexoses from dTMP and sucrose. The strategic combination of the recombinant enzymes dTMP-kinase and sucrose synthase (SuSy), and the enzymes RmlB (4,6-dehydratase), RmlC (3,5-epimerase) and RmlD (4-ketoreductase) from the biosynthetic pathway of dTDP-β-L-rhamnose was optimized. The SuSy module (dTMP-kinase, SuSy, ± RmlB) yielded the precursor dTDP-α-D-glucose (2) or the biosynthetic intermediate dTDP-6-deoxy-4-keto-α-D-glucose (3) on a 0.2-0.6 g scale with overall yields of 62% and 72%, respectively. A two-step strategy in which the SuSy module was followed by the deoxysugar module (RmlC and RmlD) resulted in the synthesis of dTDP-β-L-rhamnose (4; 24.1 μmol, overall yield: 35.9%). Substitution of RmlC by DnmU from the dTDP-β-L-daunosamine pathway of Streptomyces peucetius in this module demonstrated that DnmU acts in vitro as a 3,5-epimerase with 3 as substrate to yield 4 (32.2 μmol, overall yield: 44.7%). Chemical reduction of 3 with NaBH4 gave a mixture of the C-4 epimers dTDP-α-D-quinovose (6) and dTDP-α-D-fucose (7) in a ratio of 2:1. In summary, the modular character of the presented enzyme system provides valuable compounds for the biochemical characterization of deoxysugar pathways playing a major role in microbial producers of antibiotic and antitumour agents.
KW - Antibiotics
KW - Biocatalysis
KW - Carbohydrates
KW - Enzymes
KW - Glycoconjugates
UR - http://www.scopus.com/inward/record.url?scp=23644432034&partnerID=8YFLogxK
U2 - 10.1002/cbic.200500037
DO - 10.1002/cbic.200500037
M3 - Article
C2 - 15977277
AN - SCOPUS:23644432034
VL - 6
SP - 1423
EP - 1430
JO - ChemBioChem
JF - ChemBioChem
SN - 1439-4227
IS - 8
ER -