| RRC ID |
15412
|
| Author |
Grill S, Yamaguchi H, Wagner H, Zwahlen L, Kusch U, Mack M.
|
| Title |
Identification and characterization of two Streptomyces davawensis riboflavin biosynthesis gene clusters.
|
| Journal |
Arch Microbiol
|
| Abstract |
In Streptomyces davawensis roseoflavin is synthesized from GTP and ribulose-5-phosphate through riboflavin. As a first step towards the molecular analysis of flavin metabolism in S. davawensis the genes involved in riboflavin biosynthesis were cloned by hybridization of heterologous probes to a genomic library on a high-density colony-array. The genes ribB (riboflavin synthase, alpha-chain; EC 2.5.1.9), ribM (putative membrane protein), ribA (bifunctional GTP cyclohydrolase II/3,4-dihydroxy-2-butanone-4-phosphate synthase; EC 3.5.4.25) and ribH (lumazine synthase; EC 2.5.1.9) are organized in an operon-like cluster. Northern blot analysis of this cluster revealed two transcripts of 1.7 and 3.1 kb, respectively. The gene ribB was overexpressed in Escherichia coli. The specific riboflavin synthase activity in a cell-free extract of a recombinant strain was 0.246 nmol mg(-1 )min(-1). Overexpression of ribM enhanced the transport of riboflavin in the corresponding recombinant E. coli strain. Furthermore, overexpression of ribM increased roseoflavin sensitivity of E. coli. On another subgenomic fragment a putative S. davawensis ribG gene coding for the missing pyrimidine deaminase/reductase (EC 3.5.4.26 and EC 1.1.1.193) of the riboflavin biosynthetic pathway and ribY coding for a second (monofunctional) GTP cyclohydrolase II were identified.
|
| Volume |
188(4)
|
| Pages |
377-87
|
| Published |
2007-10-1
|
| DOI |
10.1007/s00203-007-0258-1
|
| PMID |
17541777
|
| MeSH |
Base Sequence
Biosynthetic Pathways / genetics*
Blotting, Northern
Cloning, Molecular
DNA, Bacterial / chemistry
DNA, Bacterial / genetics
Escherichia coli / genetics
Gene Order
Genomic Library
Molecular Sequence Data
Multigene Family
Nucleic Acid Hybridization
Operon
RNA, Bacterial / analysis
RNA, Bacterial / genetics
RNA, Messenger / analysis
RNA, Messenger / genetics
Recombinant Proteins / metabolism
Riboflavin / genetics*
Riboflavin Synthase / metabolism
Sequence Analysis, DNA
Streptomyces / chemistry
Streptomyces / genetics*
Streptomyces / metabolism
|
| IF |
1.884
|
| Times Cited |
27
|
|
WOS Category
|
MICROBIOLOGY
|
| Resource |
| General Microbes |
JCM 4913 |
