RRC ID 15412
著者 Grill S, Yamaguchi H, Wagner H, Zwahlen L, Kusch U, Mack M.
タイトル Identification and characterization of two Streptomyces davawensis riboflavin biosynthesis gene clusters.
ジャーナル 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.
巻・号 188(4)
ページ 377-87
公開日 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
引用数 27
WOS 分野 MICROBIOLOGY
リソース情報
一般微生物 JCM 4913