Isolation and cloning of the cutinase gene obtained from indigenous thermophiles Thermobifida fusca bacteria

Subject Areas : Environmental Microbiology

Elahe Pourfakhraei ¹ , Mehrdad Behmanesh ² , Majid Erfani Moghadam ³

1 - Department of Biophysics, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran
2 - Department of Genetics, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran
3 - Department of Biophysics, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran

Received: 2013-10-17 Accepted : 2013-11-16 Published : 2010-10-01

Keywords: Cloning, Thermobifida fusca, Cutinase,

Abstract :

Background and Objectives: Cutinase belongs to serine hydrolase family that is used for elimination of environmental pollutions due to its special structural and functional properties and applications. In this study, cutinase gene was isolated from indigenous thermophiles Thermobifida fusca and was cloned in E. coli. Material and Methods: Sampling was performed from forest soils in the north of Iran. Thermobifida fusca bacteria were isolated with suitable cultures under optimized conditions (pH 10 and T=62 °C). After primer designation, cut1 gene was isolated from bacterium and then, was cloned in pBluescript sk vector and was introduced into E. coli DH5α. The colonization was confirmed based on enzymatic digestion and sequencing.  Result: Based on BLAST software, there was 98% and 96% similarities were obtained with cut1 and cut-1.KW3, respectively.  Conclusions: Based on broad functions of the enzyme on various substrates and its ability for synthesis of various polyester compounds, this enzyme has an important role in environmental decontamination.

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