This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplementary Material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Google Scholar
Right arrow Articles by Kasai, D.
Right arrow Articles by Masai, E.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kasai, D.
Right arrow Articles by Masai, E.

 Previous Article  |  Next Article 

Journal of Bacteriology, November 2009, p. 6758-6768, Vol. 191, No. 21
0021-9193/09/$08.00+0     doi:10.1128/JB.00840-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Uncovering the Protocatechuate 2,3-Cleavage Pathway Genes{triangledown} ,{dagger}

Daisuke Kasai,1 Toshihiro Fujinami,1 Tomokuni Abe,2 Kohei Mase,2 Yoshihiro Katayama,3 Masao Fukuda,1 and Eiji Masai1*

Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata 940-2188,1 Toyota Industries Corporation, Obu, Aichi 474-8601,2 Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan3

Received 26 June 2009/ Accepted 20 August 2009

Paenibacillus sp. (formerly Bacillus macerans) strain JJ-1b is able to grow on 4-hydroxybenzoate (4HB) as a sole source of carbon and energy and is known to degrade 4HB via the protocatechuate (PCA) 2,3-cleavage pathway. However, none of the genes involved in this pathway have been identified. In this study, we identified and characterized the JJ-1b genes for the 4HB catabolic pathway via the PCA 2,3-cleavage pathway, which consisted of praR and praABEGFDCHI. Based on the enzyme activities of cell extracts of Escherichia coli carrying praI, praA, praH, praB, praC, and praD, these genes were found to code for 4HB 3-hydroxylase, PCA 2,3-dioxygenase, 5-carboxy-2-hydroxymuconate-6-semialdehyde decarboxylase, 2-hydroxymuconate-6-semialdehyde dehydrogenase, 4-oxalocrotonate (OCA) tautomerase, and OCA decarboxylase, respectively, which are involved in the conversion of 4HB into 2-hydroxypenta-2,4-dienoate (HPD). The praE, praF, and praG gene products exhibited 45 to 61% amino acid sequence identity to the corresponding enzymes responsible for the catabolism of HPD to pyruvate and acetyl coenzyme A. The deduced amino acid sequence of praR showed similarity with those of IclR-type transcriptional regulators. Reverse transcription-PCR analysis revealed that praABEGFDCHI constitute an operon, and these genes were expressed during the growth of JJ-1b on 4HB and PCA. praR-praABEGFDCHI conferred the ability to grow on 4HB to E. coli, suggesting that praEGF were functional for the conversion of HPD to pyruvate and acetyl coenzyme A. A promoter analysis suggested that praR encodes a repressor of the pra operon.

* Corresponding author. Mailing address: Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata 940-2188, Japan. Phone and fax: 81-258-47-9428. E-mail: emasai{at}vos.nagaokaut.ac.jp

{triangledown} Published ahead of print on 28 August 2009.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, November 2009, p. 6758-6768, Vol. 191, No. 21
0021-9193/09/$08.00+0     doi:10.1128/JB.00840-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»