Cloning and sequencing of the Sphingomonas (Pseudomonas) paucimobilis gene essential for the O demethylation of vanillate and syringate

Appl Environ Microbiol. 1998 Mar;64(3):836-42. doi: 10.1128/AEM.64.3.836-842.1998.

Abstract

Sphingomonas (Pseudomonas) paucimobilis SYK-6 is able to grow on 5,5'-dehydrodivanillic acid (DDVA), syringate, vanillate, and other dimeric model compounds of lignin as a sole carbon source. Nitrosoguanidine mutagenesis of S. paucimobilis SYK-6 was performed, and two mutants with altered DDVA degradation pathways were isolated. The mutant strain NT-1 could not degrade DDVA, but could degrade syringate, vanillate, and 2,2',3'-trihydroxy-3-methoxy-5,5'-dicarboxybiphenyl (OH-DDVA). Strain DC-49 could slowly assimilate DDVA, but could degrade neither vanillate nor syringate, although it could degrade protocatechuate and 3-O-methylgallate. A complementing DNA fragment of strain DC-49 was isolated from the cosmid library of strain SYK-6. The minimum DNA fragment complementing DC-49 was determined to be the 1.8-kbp insert of pKEX2.0. Sequencing analysis showed an open reading frame of 1,671 bp in this fragment, and a similarity search indicated that the deduced amino acid sequence of this open reading frame had significant similarity (60%) to the formyltetrahydrofolate synthetase of Clostridium thermoaceticum.

MeSH terms

  • Amino Acid Sequence
  • Cloning, Molecular
  • DNA, Bacterial / chemistry
  • Dealkylation
  • Gallic Acid / analogs & derivatives*
  • Gallic Acid / metabolism
  • Genes, Bacterial*
  • Lignin / metabolism
  • Molecular Sequence Data
  • Pseudomonas / genetics*
  • Vanillic Acid / metabolism*

Substances

  • DNA, Bacterial
  • Gallic Acid
  • Lignin
  • syringic acid
  • Vanillic Acid

Associated data

  • GENBANK/AB006079