Skip to main content
SpringerLink
Log in

Alginate immobilization of recombinant Escherichia coli whole cells harboring l-arabinose isomerase for l-ribulose production

  • Original Paper
  • Published:
Bioprocess and Biosystems Engineering Aims and scope Submit manuscript

Abstract

Recombinant Escherichia coli whole cells harboring Bacillus licheniformis l-arabinose isomerase (BLAI) were immobilized with alginate. The operational conditions for immobilization were optimized with response surface methodology. Optimal alginate concentration, Ca2+ concentration, and cell mass loading were 1.8% (w/v), 0.1 M, and 44.5 g L−1, respectively. The interactions between Ca2+ concentration, alginate concentration, and initial cell mass were significant. After immobilization of BLAI, cross-linking with 0.1% glutaraldehyde significantly reduced cell leakage. The half-life of immobilized whole cells was 150 days, which was 50-fold longer than that of free cells. In seven repeated batches for l-ribulose production, the productivity was as high as 56.7 g L−1 h−1 at 400 g L−1 substrate concentration. The immobilized cells retained 89% of the initial yield after 33 days of reaction. Immobilization of whole cells harboring BLAI, therefore, makes a suitable biocatalyst for the production of l-ribulose, particularly because of its high stability and low cost.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Chan LW, Heng PW (2002) Effects of aldehydes and methods of cross-linking on properties of calcium alginate microspheres prepared by emulsification. Biomaterials 23:1319–1326

    Article  CAS  Google Scholar 

  2. Chan L, Jin Y, Heng P (2002) Cross-linking mechanisms of calcium and zinc in production of alginate microspheres. Int J Pharm 242:255–258

    Article  CAS  Google Scholar 

  3. Roy I, Sharma S, Gupta MN (2004) Smart biocatalysts: design and applications. Adv Biochem Eng Biotechnol 86:159–189

    CAS  Google Scholar 

  4. Akagi M, Omae D, Tamura Y, Ueda T, Kumashiro T, Urata H (2002) A practical synthesis of l-ribose. Chem Pharm Bull 50:866–868

    Article  CAS  Google Scholar 

  5. Yun M, Moon HR, Kim HO, Choi WJ, Kim YC, Park CS, Jeong LS (2005) A highly efficient synthesis of unnatural l-sugars from d-ribose. Tetrahedron Lett 46:5903–5905

    Article  CAS  Google Scholar 

  6. Takahashi H, Iwai Y, Hitomi Y, Ikegami S (2002) Novel synthesis of l-ribose from d-mannono-1, 4-lactone. Org Lett 4:2401–2403

    Article  CAS  Google Scholar 

  7. Helanto M, Kiviharju K, Leisola M, Nyyssola A (2007) Metabolic engineering of Lactobacillus plantarum for production of l-ribulose. Appl Environ Microbiol 73:7083–7091

    Article  CAS  Google Scholar 

  8. Roh HJ, Kim P, Park YC, Choi JH (2000) Bioconversion of d-galactose into d-tagatose by expression of l-arabinose isomerase. Biotechnol Appl Biochem 31:1–4

    Article  CAS  Google Scholar 

  9. Jorgensen F, Hansen OC, Stougaard P (2004) Enzymatic conversion of d-galactose to d-tagatose: heterologous expression and characterisation of a thermostable l-arabinose isomerase from Thermoanaerobacter mathranii. Appl Microbiol Biotechnol 64:816–822

    Article  CAS  Google Scholar 

  10. Kim BC, Lee YH, Lee HS, Lee DW, Choe EA, Pyun YR (2002) Cloning, expression and characterization of l-arabinose isomerase from Thermotoga neapolitana: bioconversion of d-galactose to d-tagatose using the enzyme. FEMS Microbiol Lett 212:121–126

    CAS  Google Scholar 

  11. Lee DW, Jang HJ, Choe EA, Kim BC, Lee SJ, Kim SB, Hong YH, Pyun YR (2004) Characterization of a thermostable l-arabinose (d-galactose) isomerase from the hyperthermophilic eubacterium Thermotoga maritima. Appl Environ Microbiol 70:1397–1404

    Article  CAS  Google Scholar 

  12. Lee SJ, Lee DW, Choe EA, Hong YH, Kim SB, Kim BC, Pyun YR (2005) Characterization of a thermoacidophilic l-arabinose isomerase from Alicyclobacillus acidocaldarius: role of Lys-269 in pH optimum. Appl Environ Microbiol 71:7888–7896

    Article  CAS  Google Scholar 

  13. Prabhu P, Tiwari MK, Jeya M, Gunasekaran P, Kim IW, Lee JK (2008) Cloning and characterization of a novel l-arabinose isomerase from Bacillus licheniformis. Appl Microbiol Biotechnol 81:283–290

    Article  CAS  Google Scholar 

  14. Dische Z, Borenfreund E (1951) A new spectrophotometric method for the detection and determination of keto sugars and trioses. J Biol Chem 192:583–587

    CAS  Google Scholar 

  15. Potumarthi R, Subhakar C, Pavani A, Jetty A (2008) Evaluation of various parameters of calcium-alginate immobilization method for enhanced alkaline protease production by Bacillus licheniformis NCIM-2042 using statistical methods. Bioresour Technol 99:1776–1786

    Article  CAS  Google Scholar 

  16. Li W, Du W, Liu D (2007) Optimization of whole cell-catalyzed methanolysis of soybean oil for biodiesel production using response surface methodology. J Mol Catal B Enzym 45:122–127

    Article  CAS  Google Scholar 

  17. Anisha GS, Prema P (2008) Cell immobilization technique for the enhanced production of α-galactosidase by Streptomyces griseoloalbus. Bioresour Technol 99:3325–3330

    Article  CAS  Google Scholar 

  18. Orive G, Hernández RM, Gascón AR, Pedraz JL (2006) Encapsulation of cells in alginate gels. Method Biotechnol 22:345–355

    Article  CAS  Google Scholar 

  19. Jamuna R, Sai ST, Vora S, Ramakrishna SV (1992) Optimization of critical parameters for immobilization of yeast cells to alginate gel matrix. J Ferment Bioeng 73:319–322

    Article  CAS  Google Scholar 

  20. Alloue W, Destain J, El Medjoub T, Ghalfi H, Kabran P, Thonart P (2008) Comparison of Yarrowia lipolytica lipase immobilization yield of entrapment, adsorption, and covalent bond techniques. Appl Biochem Biotechnol 150:51–63

    Article  CAS  Google Scholar 

  21. Hung CP, Lo HF, Hsu WH, Chen SC, Lin LL (2008) Immobilization of Escherichia coli novablue γ-glutamyltranspeptidase in Ca-alginate- k -carrageenan beads. Appl Biochem Biotechnol 150:157–170

    Article  CAS  Google Scholar 

  22. Oh DK, Kim HJ, Ryu SA, Rho HJ, Kim P (2001) Development of an immobilization method of l-arabinose isomerase for industrial production of tagatose. Biotechnol Lett 23:1859–1862

    Article  CAS  Google Scholar 

  23. Novick SJ, Rozzell JD (2005) Immobilization of enzymes by covalent attachment. Method Biotechnol 17:247–271

    CAS  Google Scholar 

  24. Li S, Hu J, Liu B (2004) Use of chemically modified PMMA microspheres for enzyme immobilization. Biosystems 77:25–32

    Article  CAS  Google Scholar 

  25. Leon J, Galvan F, Vega JM (1990) Effect of immobilization on the catalytic properties of ferredoxin-nitrite reductase from Chlamydomonas reinhardtii. J Mol Catal 58:393–403

    Article  CAS  Google Scholar 

  26. Mohapatra BR (2008) Effect of immobilization on kinetic and thermodynamic characteristics of sulfide oxidase from Arthrobacter species. Prep Biochem Biotechnol 38:61–73

    Article  CAS  Google Scholar 

  27. Klibanov AM (1979) Enzyme stabilization by immobilization. Anal Biochem 93:1–25

    Article  CAS  Google Scholar 

  28. Kennedy JF, Melo EHM, Jumel K (1990) Immobilized enzymes and cells. Chem Eng Prog 86:81–89

    CAS  Google Scholar 

  29. Munjal N, Sawhney SK (2002) Stability and properties of mushroom tyrosinase entrapped in alginate, polyacrylamide and gelatin gels. Enzyme Microb Technol 30:613–619

    Article  CAS  Google Scholar 

  30. Sahin F, Demirel G, Tumturk H (2005) A novel matrix for the immobilization of acetylcholinesterase. Int J Biol Macromol 37:148–153

    Article  CAS  Google Scholar 

  31. Heath EC, Horecker BL, Smyrniotis PZ, Takagi Y (1958) Pentose fermentation by Lactobacillus plantarum. II. l-Arabinose isomerase. J Biol Chem 231:1031–1037

    CAS  Google Scholar 

  32. Yeom SJ, Ji JH, Yoon RY, Oh DK (2008) l-Ribulose production from l-arabinose by an l-arabinose isomerase mutant from Geobacillus thermodenitrificans. Biotechnol Lett 30:1789–1793

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the 21C Frontier Microbial Genomics and Applications Center Program, Ministry of Education, Science & Technology, Republic of Korea. This research was supported by a grant from the Korea Research Foundation (KRF-2007-314-D00073). This research was also supported by the 2009 KU Brain Pool of Konkuk University.

Author information

Authors and Affiliations

  1. Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul, 143-701, Korea

    Ye-Wang Zhang & Jung-Kul Lee

  2. Department of Bioscience and Biotechnology, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul, 143-701, Korea

    Ponnandy Prabhu

  3. Institute of Biomedical Science and Technology, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul, 143-701, Korea

    Jung-Kul Lee

Authors
  1. Ye-Wang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ponnandy Prabhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jung-Kul Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jung-Kul Lee.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, YW., Prabhu, P. & Lee, JK. Alginate immobilization of recombinant Escherichia coli whole cells harboring l-arabinose isomerase for l-ribulose production. Bioprocess Biosyst Eng 33, 741–748 (2010). https://doi.org/10.1007/s00449-009-0397-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00449-009-0397-7

Keywords

Search

Navigation