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PMID:17400787

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Citation

Throne-Holst, M, Wentzel, A, Ellingsen, TE, Kotlar, HK and Zotchev, SB (2007) Identification of novel genes involved in long-chain n-alkane degradation by Acinetobacter sp. strain DSM 17874. Appl. Environ. Microbiol. 73:3327-32

Abstract

Acinetobacter sp. strain DSM 17874 is capable of utilizing n-alkanes with chain lengths ranging from that of decane (C10H22) to that of tetracontane (C40H82) as a sole carbon source. Two genes encoding AlkB-type alkane hydroxylase homologues, designated alkMa and alkMb, have been shown to be involved in the degradation of n-alkanes with chain lengths of from 10 to 20 C atoms in this strain. Here, we describe a novel high-throughput screening method and the screening of a transposon mutant library to identify genes involved in the degradation of n-alkanes with C chain lengths longer than 20, which are solid at 30 degrees C, the optimal growth temperature for Acinetobacter sp. strain DSM 17874. A library consisting of approximately 6,800 Acinetobacter sp. strain DSM 17874 transposon mutants was constructed and screened for mutants unable to grow on dotriacontane (C32H66) while simultaneously showing wild-type growth characteristics on shorter-chain n-alkanes. For 23 such mutants isolated, the genes inactivated by transposon insertion were identified. Targeted inactivation and complementation studies of one of these genes, designated almA and encoding a putative flavin-binding monooxygenase, confirmed its involvement in the strain's metabolism of long-chain n-alkanes. To our knowledge, almA represents the first cloned gene shown to be involved in the bacterial degradation of long-chain n-alkanes of 32 C's and longer. Genes encoding AlmA homologues were also identified in other long-chain n-alkane-degrading Acinetobacter strains.

Links

PubMed PMC1907095 Online version:10.1128/AEM.00064-07

Keywords

Acinetobacter/enzymology; Acinetobacter/genetics; Acinetobacter/growth & development; Acinetobacter/metabolism; Alkanes/metabolism; Biodegradation, Environmental; Carbon/metabolism; Cloning, Molecular; DNA Transposable Elements; DNA, Bacterial/chemistry; DNA, Bacterial/genetics; Gene Deletion; Genetic Complementation Test; Molecular Sequence Data; Mutagenesis, Insertional; Oxygenases/genetics; Oxygenases/metabolism; Sequence Analysis, DNA; Sequence Homology, Amino Acid

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ACISP:ALMA

involved_in

GO:0043448: alkane catabolic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ACIAD:ALMA

involved_in

GO:0043448: alkane catabolic process

ECO:0000316: genetic interaction evidence used in manual assertion

UniProtKB:A5H9N6

P

Seeded From UniProt

complete

9GAMM:A5H9N6

GO:0043448: alkane catabolic process

ECO:0000315:

P

Figure 2 indicates that almA deficient mutants produce little to no protein when grown in medium containing alkanes with chain length greater than 32 whereas the wildtype is able to successfully produce protein when grown on medium containing these long chain alkanes. almA encodes AlmA, a putative flavin-binding monooxygenase, which is involved in metabolism of long chain alkanes.

complete
CACAO 4165

9GAMM:A5H9N6

involved_in

GO:0043448: alkane catabolic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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