Acrylamide synthesis using agar entrapped cells of Rhodococcus rhodochrous PA-34 in a partitioned fed batch reactor

  • Jog Raj
  • Nitya Nand Sharma
  • Shreenath Prasad
  • Tek Chand Bhalla
Original Paper


The nitrile hydratase (Nhase) induced cells of Rhodococcus rhodochrous PA-34 catalyzed the conversion of acrylonitrile to acrylamide. The cells of R. rhodochrous PA-34 immobilized in 2% (w/v) agar (1.76 mg dcw/ml agar matrix) exhibited maximum Nhase activity (8.25 U/mg dcw) for conversion of acrylonitrile to acrylamide at 10°C in the reaction mixture containing 0.1 M potassium phosphate buffer (pH 7.5), 8% (w/v) acrylonitrile and immobilized cells equivalent to 1.12 mg dcw (dry cell weight) per ml. In a partitioned fed batch reaction at 10°C, using 1.12 g dcw immobilized cells in a final volume of 1 l, a total of 372 g of acrylonitrile was completely hydrated to acrylamide (498 g) in 24 h. From the above reaction mixture 87% acrylamide (432 g) was recovered through crystallization at 4°C. By recycling the immobilized biocatalyst (six times), a total of 2,115 g acrylamide was produced.


Rhodococcus rhodochrous PA-34 Immobilized biocatalyst Agar Acrylonitrile Acrylamide Partitioned reactor 



We acknowledge the financial support in the form of Senior Research Fellowship from University Grants Commission (UGC), New Delhi, India to Mr. Jog Raj, from the Council and Scientific and Industrial Research (CSIR) to Mr. Nitya Nand Sharma and from the Department of Biotechnology, Government of India, New Delhi to Mr. Shreenath Prasad. The computational facility availed at Bioinformatics Centre, H. P. University is gratefully acknowledge.


  1. 1.
    Andhale MS, Hamde VS (1996) Isolation and screening of acrylamide producing microorganisms. Ind J Exp Biol 34:1005–1009Google Scholar
  2. 2.
    Barbotin JN, Nava-Saucedo JE (1998) Bioencapsulation of living cells by entrapment in polysaccharide gels. In: Polysaccharides—structural diversity and functional versality. Marcel Dekker, New York, pp 750–751Google Scholar
  3. 3.
    Bernet N, Thiery A, Maestracci M, Arnaud A, Rios GM, Galzy P (1987) Continuous immobilized cell reactor for amide hydrolysis. J Ind Microbiol 2:129–136CrossRefGoogle Scholar
  4. 4.
    Bui K, Arnaud A, Galzy P (1982) A new method to prepare amides by bioconversion of corresponding nitriles. Enzyme Microb Technol 4:195–197CrossRefGoogle Scholar
  5. 5.
    Chang NH, Choi KS, Lee CY (1993) Bench scale production of acrylamide using resting cells of Brevibacterium sp. CH2 in a fed batch reactor. Enzyme Microb Technol 15:979–984CrossRefGoogle Scholar
  6. 6.
    Chaplin MF, Bucke C (1990) Enzyme technology. Cambridge University Press, LondonGoogle Scholar
  7. 7.
    Fradet H, Arnaud A, Rios G, Galzy P (1985) Hydration of nitriles using a bacterial nitrile-hydratase immobilized on DEAE-cellulose. Biotechnol Bioeng XXVII:1581–1585CrossRefGoogle Scholar
  8. 8.
    Graham D, Pereira R, Varfield D, Cowan D (2000) Nitrile transformation using free and immobilized cells of thermophilic Bacillus sp Enzyme Microb Technol 26:368–373CrossRefGoogle Scholar
  9. 9.
    Hijort CM, Godtfredsen SE, Emborg C (1990) Isolation and characterization of nitrile hydratase from Rhodococcus sp J Chem Tech Biotechnol 48:217–226Google Scholar
  10. 10.
    Jallageas JC, Arnaud A, Galzy P (1980) Bioconversion of nitriles and their applications. Adv Biochem Engg 14:1–32CrossRefGoogle Scholar
  11. 11.
    Kierstan MPJ, Coughlan MP (1985) Immobilization of cells and enzymes by gel entrapment. In: Immobilized cells and enzymes—a practical approach. IRL, Oxford, pp 43–45Google Scholar
  12. 12.
    Lee CY, Hwang YB, Chang HN (1991) Acrylonitrile adaptation of Brevibacterium sp. CH1 for acrylamide production. Enzyme Microb Technol 13:53–58CrossRefGoogle Scholar
  13. 13.
    Maxwell GR (2004) Synthetic nitrogen products: a practical guide to the products and processes. Springer, Heidelberg, pp 395–396Google Scholar
  14. 14.
    Mersinger LJ, Hann EC, Cooling FB, Gavagan JE, Bassat AB, Wu S, Petrillo KL, Payne MS, Di-Cosimo R (2005) Production of acrylamide using alginate immobilized E. coli expressing Comamonas testosteroni 5- MGAM-4D nitrile hydratase. Adv Synth Catal 347:1125–1131CrossRefGoogle Scholar
  15. 15.
    Mitsubishi Rayon (2001) Enzymatic production of acrylamide. In: The application of biotechnology to industrial sustainability. OCED, France, pp 71–75Google Scholar
  16. 16.
    Nagasawa T, Shimizu H, Yamada H (1993) The superiority of the third-generation catalyst, Rhodococcus rhodochrous J1 nitrile hydratase, for industrial production of acrylamide. Appl Microbiol Biotechnol 40:189–195CrossRefGoogle Scholar
  17. 17.
    Raj J, Prasad S, Bhalla TC (2006) Rhodococcus rhodochrous PA-34 a potential catalyst for acrylamide synthesis. Process Biochem 41:1359–1363CrossRefGoogle Scholar
  18. 18.
    Yamada H, Kobayashi M (1996) Nitrile hydratase and its application to industrial production of acrylamide. Biosci Biotech Biochem 60:1391–1400CrossRefGoogle Scholar

Copyright information

© Society for Industrial Microbiology 2007

Authors and Affiliations

  • Jog Raj
    • 1
  • Nitya Nand Sharma
    • 1
  • Shreenath Prasad
    • 1
  • Tek Chand Bhalla
    • 1
  1. 1.Department of BiotechnologyHimachal Pradesh UniversityShimlaIndia

Personalised recommendations