Reference - Detail
| RRC ID | 40831 |
|---|---|
| Author | Nagayama H, Sugawara T, Endo R, Ono A, Kato H, Ohtsubo Y, Nagata Y, Tsuda M. |
| Title | Isolation of oxygenase genes for indigo-forming activity from an artificially polluted soil metagenome by functional screening using Pseudomonas putida strains as hosts. |
| Journal | Appl Microbiol Biotechnol |
| Abstract |
Metagenomes contain the DNA from many microorganisms, both culturable and non-culturable, and are a potential resource of novel genes. In this study, a 5.2-Gb metagenomic DNA library was constructed from a soil sample (artificially polluted with four aromatic compounds, i.e., biphenyl, phenanthrene, carbazole, and 3-chlorobenzoate) in Escherichia coli by using a broad-host-range cosmid vector. The resultant library was introduced into naphthalene-degrading Pseudomonas putida-derived strains having deficiencies in their naphthalene dioxygenase components, and indigo-forming clones on the indole-containing agar plates were screened. Cosmids isolated from 29 positive clones were classified by their various properties (original screening hosts, hosts showing indigo-forming activity, and digestion patterns with restriction enzymes), and six representative cosmids were chosen. Sequencing and in vitro transposon mutagenesis of the six cosmids resulted in the identification of genes encoding putative class B and D flavoprotein monooxygenases, a multicomponent hydroxylase, and a reductase that were responsible for the indigo-forming activity in the host cells. Among them, the genes encoding the multicomponent hydroxylase were demonstrated to be involved in phenol degradation. Furthermore, two genes encoding ring-cleavage dioxygenases were also found adjacent to the genes responsible for the indigo formation, and their functions were experimentally confirmed. |
| Volume | 99(10) |
| Pages | 4453-70 |
| Published | 2015-5-1 |
| DOI | 10.1007/s00253-014-6322-2 |
| PMID | 25573469 |
| MeSH | Bacteria / classification Bacteria / enzymology* Bacteria / genetics Bacteria / isolation & purification Bacterial Proteins / genetics* Bacterial Proteins / metabolism Biodegradation, Environmental Indigo Carmine / metabolism* Metagenome* Molecular Sequence Data Oxygenases / genetics* Oxygenases / metabolism Pseudomonas putida / genetics* Pseudomonas putida / metabolism Soil Microbiology* Soil Pollutants / metabolism* |
| IF | 3.53 |
| Times Cited | 18 |
| WOS Category | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| Altmetric score |
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 2 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| General Microbes | JCM 18452 |