Surfactant-mediated permeabilization of Pseudomonas putida KT2440 and use of the immobilized permeabilized cells in biotransformation |
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| Institution: | 1. Department of Pharmaceutical Technology (Biotechnology), National Institute of Pharmaceutical Education and Research, Sector-67, S.A.S. Nagar, 160062, Punjab, India;2. School of Engineering, Massey University, Private Bag11222, Palmerston North, New Zealand;1. Institute for Innovative Learning, Mahidol University, Nakhon Pathom 73170, Thailand;2. Department of Biochemistry and Center for Excellence in Protein and Enzyme Technology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand;3. Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand;4. Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand;5. Department of Biomolecular Science and Engineering, School of Biomolecular Science & Engineering, Wangchan Valley, Rayong Vidyasirimedhi Institute of Science and Technology (VISTEC), 21210, Thailand;1. Energy and Environment Fusion Technology Center (E2FTC), Department of Energy Science and Technology (DEST), Myongji University, 116 Myongji-ro, Cheoin-gu, Yongin-si, Gyeonggi-do, 17058, Republic of Korea;2. Division of Bioscience and Bioinformatics, Myongji University, 116 Myongji-ro, Cheoin-gu, Yongin-si, Gyeonggi-do, 17058, Republic of Korea;1. University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh 160014, India;2. Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector-67, S.A.S. Nagar 160062, Punjab, India;3. Department of Pharmaceutical Engineering and Technology, Indian Institute of Technology (BHU), India;4. Amity University, Sector 82A, Mohali, Punjab 140306, India |
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| Abstract: | Surfactants were used to permeabilize cells of Pseudomonas putida KT2440 so as to maximize retention of the arginine deiminase (ADI) activity within the treated cells. The surfactants cetyltrimethylammoniumbromide (CTAB), sodium dodecyl sulfate (SDS) and Triton X100 were tested separately. Statistical models were developed for the effects on the ADI activity of the following factors: the concentration of the surfactant, the length of the treatment period and the concentration of the cells. For all surfactants, the concentration of cells was the most significant factor in influencing permeabilization. All permeabilization treatments used mild conditions (pH 7, 37 °C). The permeabilized cells were immobilized in alginate beads for the biotransformation of arginine to citrulline. The optimal conditions for immobilization and biotransformation were as follows: 2% (w/v, g/100 mL) sodium alginate, 100 g/L of treated cells, 40 mM arginine, pH 6.0, a temperature of 35 °C and an agitation speed of 150 rpm. The immobilized biocatalyst retained nearly 90% of its initial activity after nine cycles of repeated use in batch operations. In contrast, the freely suspended cells were barely active after the second use cycle. |
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| Keywords: | Cell permeabilization Arginine deiminase Whole-cell immobilization Biotransformation Citrulline |
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