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Biotransformation of amide using Bacillus sp.: isolation strategy, strain characteristics and enzyme immobilization

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Abstract

Amidase hydrolases/acyltransferases are of considerable industrial interest due to potential applications in the production of useful hydroxamic acids. The test strain, capable of acetamide degradation, was isolated by an enrichment technique with acetamide as sole source of nitrogen. Based on morphology, physiological tests and biochemical tests, this isolate was identified as Bacillus sp. and on the basis of 16S Ribosomal ribonucleic acid sequence, a phylogenetic tree was drawn and was identified as Bacillus megaterium. Resting cells containing active acyltransferase enzyme were prepared and immobilized in the gel beads of sodium alginate, agar, polyacrylamide and polyvinyl alcohol–alginate. The beads were tested for acyltransferase using Iron (III) chloride reagents at 55°C and were found to be affected by substrate concentration, type of buffer, buffer pH and reaction temperature. These factors were optimized using sodium alginate immobilized beads. This study proved useful in understanding the technique of immobilization of acyltransferase enzyme, its operational stability and its importance in the synthesis of hydroxamic acid.

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Acknowledgments

This work was supported by Jaipur Engineering College and Research Centre (JECRC, Jaipur, India). Authors are thankful to Mr. Arpit Agarwal (Director, JECRC), Prof. Reena Mathur (HOD, Dept. of Zoology, University of Rajasthan, Jaipur), Dr. C.P. Malik from Jaipur National University, India and Dr. S.P. Chaurasia, Chem. Eng Dept., MNIT, Jaipur for their kind suggestions and helpful discussion. Authors also wish to thank Prof. Santra of Therachem Medilab, Jaipur (India) for sample purification by preparative HPLC and NCCS, Pune (India) for gene sequencing.

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Authors and Affiliations

  1. Department of Zoology, University of Rajasthan, Jaipur, India

    M. Sogani, N. Mathur & P. Sharma

  2. IIS University, Mansarovar, Jaipur, India

    P. Bhatnagar

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  1. M. Sogani
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  2. N. Mathur
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  3. P. Bhatnagar
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  4. P. Sharma
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Correspondence to M. Sogani.

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Sogani, M., Mathur, N., Bhatnagar, P. et al. Biotransformation of amide using Bacillus sp.: isolation strategy, strain characteristics and enzyme immobilization. Int. J. Environ. Sci. Technol. 9, 119–127 (2012). https://doi.org/10.1007/s13762-011-0005-7

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  • DOI: https://doi.org/10.1007/s13762-011-0005-7

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