The ribR gene encodes a monofunctional riboflavin kinase which is involved in regulation of the Bacillus subtilis riboflavin operon

I. M. Solovieva, R. A. Kreneva, D. J. Leak, D. A. Perumov

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

A 3.5 kb EcoRI-BamHI fragment of Bacillus subtilis chromosomal DNA carrying the ribR gene, involved in regulation of the B. subtilis riboflavin operon, was cloned in the B. subtilis-Escherichia coli shuttle vector pCB20. DNA sequence analysis of this fragment revealed several ORFs, one of which encodes a polypeptide of 230 amino acids with up to 45% sequence identity with FAD synthetases from a number of micro-organisms, such as Corynebacterium ammoniagenes, E. coli and Pseudomonas fluorescens, and also to the ribC gene product of B. subtilis. The ribR gene was amplified by PCR, cloned and expressed in E. coli. Measurement of flavokinase activity in cell extracts demonstrated that ribR encodes a monofunctional flavokinase which converts riboflavin into FMN but not to FAD, and is specific for the reduced form of riboflavin.

Original languageEnglish
Pages (from-to)67-73
Number of pages7
JournalMicrobiology
Volume145
Issue number1
DOIs
Publication statusPublished - 1 Jan 1999

Fingerprint

riboflavin kinase
Riboflavin
Operon
Bacillus subtilis
Escherichia coli
FMN adenylyltransferase
Genes
Flavin Mononucleotide
Corynebacterium
Pseudomonas fluorescens
Genetic Vectors
Flavin-Adenine Dinucleotide
Cell Extracts
DNA Sequence Analysis
Open Reading Frames
Amino Acids
Polymerase Chain Reaction
Peptides
DNA

Keywords

  • Bacillus subtilis
  • Flavinogenesis
  • Flavokinase
  • Operon regulation
  • Riboflavin

ASJC Scopus subject areas

  • Microbiology

Cite this

The ribR gene encodes a monofunctional riboflavin kinase which is involved in regulation of the Bacillus subtilis riboflavin operon. / Solovieva, I. M.; Kreneva, R. A.; Leak, D. J.; Perumov, D. A.

In: Microbiology, Vol. 145, No. 1, 01.01.1999, p. 67-73.

Research output: Contribution to journalArticle

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AU - Leak, D. J.

AU - Perumov, D. A.

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N2 - A 3.5 kb EcoRI-BamHI fragment of Bacillus subtilis chromosomal DNA carrying the ribR gene, involved in regulation of the B. subtilis riboflavin operon, was cloned in the B. subtilis-Escherichia coli shuttle vector pCB20. DNA sequence analysis of this fragment revealed several ORFs, one of which encodes a polypeptide of 230 amino acids with up to 45% sequence identity with FAD synthetases from a number of micro-organisms, such as Corynebacterium ammoniagenes, E. coli and Pseudomonas fluorescens, and also to the ribC gene product of B. subtilis. The ribR gene was amplified by PCR, cloned and expressed in E. coli. Measurement of flavokinase activity in cell extracts demonstrated that ribR encodes a monofunctional flavokinase which converts riboflavin into FMN but not to FAD, and is specific for the reduced form of riboflavin.

AB - A 3.5 kb EcoRI-BamHI fragment of Bacillus subtilis chromosomal DNA carrying the ribR gene, involved in regulation of the B. subtilis riboflavin operon, was cloned in the B. subtilis-Escherichia coli shuttle vector pCB20. DNA sequence analysis of this fragment revealed several ORFs, one of which encodes a polypeptide of 230 amino acids with up to 45% sequence identity with FAD synthetases from a number of micro-organisms, such as Corynebacterium ammoniagenes, E. coli and Pseudomonas fluorescens, and also to the ribC gene product of B. subtilis. The ribR gene was amplified by PCR, cloned and expressed in E. coli. Measurement of flavokinase activity in cell extracts demonstrated that ribR encodes a monofunctional flavokinase which converts riboflavin into FMN but not to FAD, and is specific for the reduced form of riboflavin.

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