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Gene structures and properties of enzymes of the plasmid-encoded nicotine catabolism of Arthrobacter nicotinovorans

Schenk, Susann and Hoelz, André and Krauβ, Beate and Decker, Karl (1998) Gene structures and properties of enzymes of the plasmid-encoded nicotine catabolism of Arthrobacter nicotinovorans. Journal of Molecular Biology, 284 (5). pp. 1323-1339. ISSN 0022-2836. https://resolver.caltech.edu/CaltechAUTHORS:20200407-130101803

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Abstract

Arthrobacter nicotinovorans is a Gram-positive aerobic soil bacterium able to grow on nicotine as its sole source of carbon and nitrogen. The initial steps of nicotine catabolism are catalyzed by nicotine dehydrogenase, the l- and d-specific 6-hydroxynicotine oxidases, and ketone dehydrogenase. The genes encoding these enzymes reside on a 160 kb plasmid, pAO1. The cccDNA of this plasmid was isolated in high purity and reasonable yield. It served as template material for the construction of a λ-phage DNA library of the plasmid. The genes coding for 6-hydroxy-l-nicotine oxidase and for the subunits of the heterotrimeric ketone dehydrogenase were identified, subcloned and sequenced. The 6-hlno gene was identified as a 1278 bp open reading frame; its regulatory elements were also recognized. The derived primary structure of the monomer of apo-6-hydroxy-l-nicotine oxidase (46,264.5 Da) agrees with the data obtained by partial amino acid sequencing. 6-Hydroxy-l-nicotine oxidase and 6-hydroxy-d-nicotine oxidase were expressed in Escherichia coli and obtained in a state of high purity and crystallized. Ketone dehydrogenase (KDH) was found to be a heterotrimer with subunits of molecular mass 89,021.71, 26,778.65 and 17,638.88. The genes of KDH-A and KDH-B are juxtaposed; the A of the stop codon of KDH-A is used in the start codon of KDH-B, eliciting a frame shift. KDH-C is separated from KDH-A by 281 bp.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1006/jmbi.1998.2227DOIArticle
ORCID:
AuthorORCID
Hoelz, André0000-0003-1726-0127
Additional Information:© 1998 Academic Press. Received 22 May 1998, Revised 24 August 1998, Accepted 3 September 1998. We thank Dr R. Maier (Freiburg) for his support in the use of the pulsed-field gel electrophoresis, Dr C. Menéndez (Freiburg) for providing the transposase A gene, Dr M. Moczko (Freiburg) for the translocase of outer membrane 70 probe. We are indebted to Dr Schlitz (Freiburg) for performing the gas-phase peptide sequencing. This work was supported by grants from the Deutsche Forschungsgemeinschaft, Bonn, through SFB 206 and De 113/34-1 and by Fonds der Chemischen Industrie, Frankfurt-M. Accession numbers: The DNA and derived protein sequences are deposited at the EMBL Nucleotide Sequence Database, accession numbers AJ 223391 (hlno), AJ 001136 (kdhA), AJ 001137 (kdhB) and AJ 001138 (kdhC).
Funders:
Funding AgencyGrant Number
Deutsche Forschungsgemeinschaft (DFG)SFB 206
Deutsche Forschungsgemeinschaft (DFG)De 113/34-1
Fonds der Chemischen IndustrieUNSPECIFIED
Subject Keywords:6-hydroxy-d-nicotine oxidase; 6-hydroxy-l-nicotine oxidase; ketone dehydrogenase; λ-phage library; sequence homologies
Issue or Number:5
Record Number:CaltechAUTHORS:20200407-130101803
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20200407-130101803
Official Citation:Susann Schenk, André Hoelz, Beate Krauβ, Karl Decker, Gene structures and properties of enzymes of the plasmid-encoded nicotine catabolism of Arthrobacter nicotinovorans11Edited by J. Karn, Journal of Molecular Biology, Volume 284, Issue 5, 1998, Pages 1323-1339, ISSN 0022-2836, https://doi.org/10.1006/jmbi.1998.2227. (http://www.sciencedirect.com/science/article/pii/S0022283698922276)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:102387
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:07 Apr 2020 20:15
Last Modified:07 Apr 2020 20:15

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