Expression of Alkane Hydroxylase from
            <i>Acinetobacter</i>
            sp. Strain ADP1 Is Induced by a Broad Range of n- Alkanes and Requires the Transcriptional Activator AlkR

Ratajczak, Andreas; Geißdörfer, Walter; Hillen, Wolfgang

doi:10.1128/jb.180.22.5822-5827.1998

Cited by 88 publications

(47 citation statements)

References 24 publications

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“…It was also shown using transcriptional fusion assays that the alkB gene from Acintetobacter sp. strain ADP1 was induced to varying extents by a range of n-alkanes (Ratajczak et al, 1998b). In addition, our results indicate that although expression of the alkB gene is induced by alkanes, this induction is also highly influenced by temperature.…”

Section: Discussionmentioning

confidence: 56%

“…By contrast, the genes encoding the alkB complex in Acinetobacter sp. APD1 appear to be arranged in a totally irregular fashion (Ratajczak et al, 1998b). Alkane degradation in Acinetobacter is highly dependent on the expression of at least five genes, with alkR known to regulate transcription of the entire gene cluster (Ratajczak et al, 1998a).…”

Section: Introductionmentioning

confidence: 99%

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The degradation of n-hexadecane in soil by thermophilic geobacilli

Marchant¹,

Sharkey²,

Banat³

et al. 2006

FEMS Microbiology Ecology

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Soil microcosms have been used to demonstrate the ability of indigenous soil thermophiles to degrade effectively a representative alkane (n-hexadecane). A fragment of the alkane mono-oxygenase gene (alkB) was amplified from thermophilic Geobacillus thermoleovorans strain T70 by PCR using degenerate primers. The amplicon demonstrated 96% sequence similarity with the alkB gene from Rhodococcus erythropolis. Critical controls ensured that the positive PCR signal detected was not a result of mesophilic soil organisms. A reverse transcription PCR (RT-PCR) assay was developed to determine if expression of the gene was inducible in the presence of an alkane or constitutively expressed in soil. In the presence of n-hexadecane, expression of the alkane mono-oxygenase gene was induced in pure cultures and soil samples and was dependent on temperature. No positive RT-PCR signal was detected at mesophilic growth temperatures either in pure cultures or in soil microcosms, whereas at 55 degrees C positive RT-PCR signals were obtained for both pure cultures of T70 and soil samples. Many different amplicons of the alkB gene fragment were obtained from the soil used in the microcosms. Thirty cloned fragments yielded 27 different sequences showing 85-96% sequence similarity with the alkB sequence of T70. To establish that the amplified alkB gene sequences from soil were derived from thermophilic geobacilli, additional strains were isolated on a selective medium containing n-hexadecane as sole carbon source. The 16S rRNA gene sequences were determined to identify the 50 isolates obtained (G. thermoleovorans, 27; G. caldoxylosilyticus, 17; G. pallidus, 2; G. toebiii, 1; Geobacillus sp., 3) representing 18 different strains and alkB gene sequences determined and deposited with the European Bioinformatics Institute.

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Section: Discussionmentioning

confidence: 56%

Section: Introductionmentioning

confidence: 99%

The degradation of n-hexadecane in soil by thermophilic geobacilli

Marchant¹,

Sharkey²,

Banat³

et al. 2006

FEMS Microbiology Ecology

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“…As a positive control, plasmid pWH968, containing the Acinetobacter sp. ADP1 alkRM and rubAB operons, was used (Ratajczak et al, 1998b). Both plasmids complemented WH405 for growth on dodecane, proving that AlkM (A. calcoaceticus EB104) encodes an alkane hydroxylase.…”

Section: Probe Bmentioning

confidence: 99%

Molecular screening for alkane hydroxylase genes in Gram‐negative and Gram‐positive strains

Smits

Röthlisberger

Witholt

et al. 1999

Environmental Microbiology

196

211

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We have developed highly degenerate oligonucleotides for polymerase chain reaction (PCR) amplification of genes related to the Pseudomonas oleovorans GPo1 and Acinetobacter sp. ADP1 alkane hydroxylases, based on a number of highly conserved sequence motifs. In all Gram-negative and in two out of three Gram-positive strains able to grow on medium- (C6-C11) or long-chain n-alkanes (C12-C16), PCR products of the expected size were obtained. The PCR fragments were cloned and sequenced and found to encode peptides with 43.2-93.8% sequence identity to the corresponding fragment of the P. oleovorans GPo1 alkane hydroxylase. Strains that were unable to grow on n-alkanes did not yield PCR products with homology to alkane hydroxylase genes. The alkane hydroxylase genes of Acinetobacter calcoaceticus EB104 and Pseudomonas putida P1 were cloned using the PCR products as probes. The two genes allow an alkane hydroxylase-negative mutant of Acinetobacter sp. ADP1 and an Escherichia coli recombinant containing all P. oleovorans alk genes except alkB, respectively, to grow on n-alkanes, showing that the cloned genes do indeed encode alkane hydroxylases.

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“…Alkane monooxygenases are the key enzymes found in bacterial alkane degradation pathways and, in general, they catalyze the initial terminal oxidation of the alkane substrate to a 1-alkanol, usually as part of a three component alkane hydroxylase complex consisting of a particulate, integral-membrane alkane monooxygenase, and two soluble proteins, rubredoxin and rubredoxin reductase. The alkane monooxygenases from only a small number of Gram-negative bacteria, namely Pseudomonas (Pp alkB; C5^C12 n-alkanes) [17,18] and Acinetobacter (Ac alkM; C10^C20 n-alkanes) [19,20], have been genetically characterized in detail. We have recently found and characterized multiple alkane hydroxylase systems in two Rhodococcus strains (R. erythropolis NRRL B-16531 and Rhodococcus sp.…”

Section: Introductionmentioning

confidence: 99%

Prevalence of alkane monooxygenase genes in Arctic and Antarctic hydrocarbon-contaminated and pristine soils

Whyte¹

2002

FEMS Microbiology Ecology

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The prevalence of four alkane monooxygenase genotypes (Pseudomonas putida GPo1, Pp alkB; Rhodococcus sp. strain Q15, Rh alkB1 and Rh alkB2; and Acinetobacter sp. strain ADP-1, Ac alkM) in hydrocarbon-contaminated and pristine soils from the Arctic and Antarctica were determined by both culture-independent (PCR hybridization analyses) and culture-dependent (colony hybridization analyses) molecular methods, using oligonucleotide primers and DNA probes specific for each of the alk genotypes. PCR hybridization of total soil community DNA detected the rhodococcal alkB genotypes in most of the contaminated (Rh alkB1, 18/20 soils; Rh alkB2, 13/20) and many pristine soils (Rh alkB1, 9/10 soils; Rh alkB2, 7/10), while Pp alkB was generally detected in the contaminated soils (15/20) but less often in pristine soils (5/10). Ac alkM was rarely detected in the soils (1/30). The colony hybridization technique was used to determine the prevalence of each of the alk genes and determine their relative abundance in culturable cold-adapted (5 ‡C) and mesophilic populations (37 ‡C) from eight of the polar soils. The cold-adapted populations, in general, possessed relatively higher percentages of the Rh alkB genotypes (Rh alkB1, 1.9% (0.55); Rh alkB2, 2.47% (0.89)), followed by the Pp alkB (1.13% (0.50)), and then the Ac alkM (0.53% (0.36)). The Rh alkB1 genotype was clearly more prevalent in culturable cold-adapted bacteria (1.9% (0.55)) than in culturable mesophiles (0.41 (0.55)), suggesting that cold-adapted bacteria are the predominant organisms possessing this genotype. Overall, these results indicated that (i) Acinetobacter spp. are not predominant members of polar alkane degradative microbial communities, (ii) Pseudomonas spp. may become enriched in polar soils following contamination events, and (iii) Rhodococcus spp. may be the predominant alkane-degradative bacteria in both pristine and contaminated polar soils. ß

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Expression of Alkane Hydroxylase from Acinetobacter sp. Strain ADP1 Is Induced by a Broad Range of n- Alkanes and Requires the Transcriptional Activator AlkR

Cited by 88 publications

References 24 publications

The degradation of n-hexadecane in soil by thermophilic geobacilli

The degradation of n-hexadecane in soil by thermophilic geobacilli

Molecular screening for alkane hydroxylase genes in Gram‐negative and Gram‐positive strains

Prevalence of alkane monooxygenase genes in Arctic and Antarctic hydrocarbon-contaminated and pristine soils

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