Cloning and sequencing of the Sphingomonas (Pseudomonas) paucimobilis gene essential for the O demethylation of vanillate and syringate.
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.
Nishikawa
S
S
New Products & Technology Laboratory, Cosmo Research Institute, Saitama, Japan. seiji-nk@ja2.so-net.or.jp
Sonoki
T
T
Kasahara
T
T
Obi
T
T
Kubota
S
S
Kawai
S
S
Morohoshi
N
N
Katayama
Y
Y
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry; D003640:Dealkylation; D005707:Gallic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D005798:Genes, Bacterial; D008031:Lignin / Q000378:metabolism; D008969:Molecular Sequence Data; D011549:Pseudomonas / Q000235:genetics*; D014641:Vanillic Acid / Q000378:metabolism*
D004269:DNA, Bacterial; D005707:Gallic Acid; D008031:Lignin; C001945:syringic acid; D014641:Vanillic Acid