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Appl Environ Microbiol, March 1998, p. 836-842, Vol. 64, No. 3
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Cloning and Sequencing of the Sphingomonas (Pseudomonas) paucimobilis Gene Essential for the O Demethylation of Vanillate and Syringate

Seiji Nishikawa,1,* Tomonori Sonoki,2 Tatsuhide Kasahara,2 Takahiro Obi,2 Shoko Kubota,2 Shinya Kawai,3 Noriyuki Morohoshi,3 and Yoshihiro Katayama2

New Products & Technology Laboratory, Cosmo Research Institute, 1134-2 Gongendo Satte Saitama 340-01,1 and Graduate School of Bio-applications & System Engineering,2 and Laboratory of Wood Chemistry, Faculty of Agriculture,3 Tokyo Noko University, Fuchu, Tokyo 183, Japan

Received 29 May 1997/Accepted 22 December 1997

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.

* Corresponding author. Mailing address: New Products & Technology Laboratory, Cosmo Research Institute, 1134-2 Gongendo Satte, Saitama 340-01, Japan. Phone: 81-480-42-2211. Fax: 480-42-3790. E-mail: seiji-nk{at}ja2.so-net.or.jp.




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