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| Content Provider | PubMed Central |
|---|---|
| Author | Silva, Elisabete Marques, Ana Rita Fialho, Arsénio Mendes Granja, Ana Teresa Isabel, Sá-correia |
| Copyright Year | 2005 |
| Abstract | The commercial gelling agent gellan is a heteropolysaccharide produced by Sphingomonas elodea ATCC 31461. In this work, we carried out the biochemical characterization of the enzyme encoded by the first gene (rmlA) of the rml 4-gene cluster present in the 18-gene cluster required for gellan biosynthesis (gel cluster). Based on sequence homology, the putative rml operon is presumably involved in the biosynthesis of dTDP-rhamnose, the sugar necessary for the incorporation of rhamnose in the gellan repeating unit. Heterologous RmlA was purified as a fused His6-RmlA protein from extracts prepared from Escherichia coli IPTG (isopropyl-β-d-thiogalactopyranoside)-induced cells, and the protein was proven to exhibit dTDP-glucose pyrophosphorylase (Km of 12.0 μM for dTDP-glucose) and UDP-glucose pyrophosphorylase (Km of 229.0 μM for UDP-glucose) activities in vitro. The N-terminal region of RmlA exhibits the motif G-X-G-T-R-X2-P-X-T, which is highly conserved among bacterial XDP-sugar pyrophosphorylases. The motif E-E-K-P, with the conserved lysine residue (K163) predicted to be essential for glucose-1-phosphate binding, was observed. The S. elodea ATCC 31461 UgpG protein, encoded by the ugpG gene which maps outside the gel cluster, was previously identified as the UDP-glucose pyrophosphorylase involved in the formation of UDP-glucose, also required for gellan synthesis. In this study, we demonstrate that UgpG also exhibits dTDP-glucose pyrophosphorylase activity in vitro and compare the kinetic parameters of the two proteins for both substrates. DNA sequencing of ugpG gene-adjacent regions and sequence similarity studies suggest that this gene maps with others involved in the formation of sugar nucleotides presumably required for the biosynthesis of another cell polysaccharide(s). |
| Related Links | http://dx.doi.org/10.1128/aem.71.8.4703-4712.2005 |
| Ending Page | 4712 |
| Page Count | 10 |
| Starting Page | 4703 |
| File Format | |
| ISSN | 00992240 |
| e-ISSN | 10985336 |
| Journal | Applied and Environmental Microbiology |
| Issue Number | 8 |
| Volume Number | 71 |
| Language | English |
| Publisher | American Society for Microbiology |
| Publisher Date | 2005-08-01 |
| Access Restriction | Open |
| Rights Holder | American Society for Microbiology |
| Subject Keyword | Biotechnology Food Science Ecology Applied Microbiology and Biotechnology Research in Higher Education |
| Content Type | Text |
| Resource Type | Article |
| Subject | Ecology Food Science Applied Microbiology and Biotechnology Biotechnology |
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