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Cloning and Characterization of a Plasmid Encoded ACC Deaminase from an Indigenous Pseudomonas fluorescens FY32

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Abstract

In addition to the characterized mechanisms responsible for many direct effects of plant growth promoting bacteria (PGPB) on plants, it has been suggested that a number of PGPB contain the enzyme ACC deaminase that catalyzes degradation of 1-aminocyclopropane-1-carboxylic acid (ACC), the immediate precursor of ethylene, into α-ketobutyrate and ammonia. As part of an effort to obtain an ACC deaminase encoding gene from a collection of soil samples, only one bacterial isolate, Pseudomonas fluorescens FY32 was capable of growing on ACC as a sole source of nitrogen. The ACC deaminase gene was amplified from the above isolate by polymerase chain reaction (PCR) giving an expected DNA fragment, 1017 bp. Sequence analysis of the fragment showed that it was highly homologous (94% and 98% identities at nucleotide and amino acid levels, respectively) to the previously characterized acdS gene from Pseudomonas sp. 6G5. Furthermore, fusion of the ACC deaminase ORF with lacZ gene resulted in the expression of active enzyme in Escherichia coli. In addition, further analyses revealed that the acdS gene was plasmid-encoded so that a large plasmid (pFY32) with almost 50 kb in size was identified from this bacterium. Furthermore, transfer of pFY32 into E. coli DH5α proved its ACC deaminase activity. This result was in accordance with previous reports suggesting horizontal transfer of the acdS gene. However, it needs more investigation to identify whether this pFY32 plasmid has undergone lateral gene transfer during the evolutionary process.

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Authors and Affiliations

  1. Soil Biology and Biotechnology Department, University of Tabriz, P.O. Box 51664, Tabriz, Iran

    Davoud Farajzadeh & Naser Aliasgharzad

  2. Industrial and Environmental Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

    Davoud Farajzadeh & Bagher Yakhchali

  3. Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK, 74078, USA

    Nemat Sokhandan Bashir

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  1. Davoud Farajzadeh
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  2. Naser Aliasgharzad
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  3. Nemat Sokhandan Bashir
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  4. Bagher Yakhchali
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Correspondence to Davoud Farajzadeh.

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Farajzadeh, D., Aliasgharzad, N., Sokhandan Bashir, N. et al. Cloning and Characterization of a Plasmid Encoded ACC Deaminase from an Indigenous Pseudomonas fluorescens FY32. Curr Microbiol 61, 37–43 (2010). https://doi.org/10.1007/s00284-009-9573-x

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