Cloning and expression of rhl AB operon under the control of tac promoter in E. coli

Document Type : Research Article

Authors

• jamshid Raheb ¹
• seyed abolghassem mohammadi bondarkhilli ¹
• Mehdi MohMMADI ²
• Kambiz Akbari ²
• Hossein Shahbani ³
• Arefeh Alipour ⁴

¹ National Institute of Genetics and Biotechnology Engineering, Research Institute of Medical Biotechnology, Department of Molecular Medicine

² Natinal Institute of Genetic Engineering and Biotechnology

³ National Institute of Genetic Engineering and Biotechnology

⁴ National Institute of Genetic Rngineering and Biotechnology

Abstract

Today, efforts go towards the replacement of chemical surfactants by natural biological biosurfactants (biosurfactant), as these materials are not carcinogenic and highly compatibile with the environment. One of the main classes of biosurfactants is rhamnose containing glycolipid biosurfactant (rhamnolipids). This type of biosurfactants can be applied in many industries such as oil industry, pharmaceutical industry, food industry and esc. In the present study, with elimination of regulatory elements of monorhamnolipid operon, the suitable construct was made and transformed to E. coli BL12 and E. coli DH5α competent cells. Then the production of monorhamnolipid in recombinant E. coli BL12 harboring pET 12a vector and mono and di-rhamnolipid biosurfactant in recombinant E. coli DH5α harboring pTrc 99A evaluated by CTAB plate and thin layer chromatography (TLC). The relative production of mono and dirhamnolipid biosurfactant was determined by oil spreading method. Finally, the efficiency of the extracted biosurfactant was verified by E12 method in different salinity and pH conditions.

Keywords

• Biosurfactant
• Rhamnolipid
• E. coli BL12
• E. coli DH5α
• transformation

Main Subjects

• Molecular Microbiology

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