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References

  1. Andrews BA, Pyle DL, Asenjo JA. 1994. The effects of pH and ionic strength on the partitioning of four proteins in reverse micelle systems. Biotechnol. Bioeng. 43: 1052-1058.
    Pubmed CrossRef
  2. Beg QK, Kapoor M, Mahajan L, Hoondal GS. 2001. Microbial xylanases and their industrial applications: a review. Appl. Microbiol. Biotechnol. 56: 326-338.
    Pubmed CrossRef
  3. Bera MB, Panesar PS, Panesar R, Singh B. 2008. Application of reverse micelle extraction process for amylase recovery using response surface methodology. Bioprocess Biosyst. Eng. 31: 379-384.
    Pubmed CrossRef
  4. Bhatti HN, Rashid MH, Nawaz R, Asgher MM, Perveen R, Jabbar A. 2007. Purication and characterization of a novel glucoamylase from Fusarium solani. Food Chem. 103: 338-343.
    CrossRef
  5. Chen YL, Su CK, Chiang BH. 2006. Optimization of reversed micellar extraction of chitosanase produced by Bacillus cereus. Process Biochem. 41: 752-758.
    CrossRef
  6. Cortez EV Jr AP, Felipe MGA, Roberto IC, Vitolo M. 2004. Optimized extraction by cetyltrimethyl ammonium bromide reversed micelles of xylose reductase and xylitol dehydrogenase from Candida guilliermondii homogenate. J. Chromatogr. B 807: 47-54.
    Pubmed CrossRef
  7. Dean A, Voss D. 1999. Design and Analysis of Experiments. Springer-Verlag New York.
    CrossRef
  8. Ferreira LFP, Taqueda ME, Vitoloc M, Converti A, Pessoa Jr A. 2005. Liquid–liquid extraction of commercial glucose oxidase by reversed micelles. J. Biotechnol. 116: 411-416.
    Pubmed CrossRef
  9. George DA, Stuckey DC. 2010. Extraction of monoclonal antibodies (IgG1) using anionic and anionic/nonionic reverse micelles. Biotechnol. Progress 26: 1352-1360.
    Pubmed CrossRef
  10. Ghildyal NP, Ramakrishna M, Lonsane BK, Karanth NG. 1991. Efficient and simple extraction of mouldy bran in a pulsed column extractor for recovery of amyloglucosidase in concentrated form. Process Biochem. 26: 235-241.
    CrossRef
  11. Hasmann FA, Gurpilhares DB, Roberto IC, Pessoa Jr A. 2007. Response surface methodology for the evaluation of glucose-6-phosphate dehydrogenase enrichment process by soybean lecithin reversed micelles. J. Chromatogr. B 847: 262-266.
    Pubmed CrossRef
  12. Hayes DG, Marchio C. 1998. Expulsion of proteins from water-in-oil microemulsion by treatment with cosurfactant. Biotechnol. Bioeng. 59: 557-566.
    CrossRef
  13. Hemavathi AB, Hebbar HU, Raghavarao KSMS. 2010. Mixed reverse micellar systems for extraction and purification of βglucosidase. Sep. Purif. Technol. 71: 263-268.
    CrossRef
  14. Ikasari L, Mitchell DA. 1996. Leaching and characterization of Rhizopus oligosporus acid protease from solid-state fermentation. Enzyme Microb. Technol. 19: 171-175.
    CrossRef
  15. Kalisz HM. 1988. Microbial proteinases. Adv. Biochem. Eng. Biotechnol. 36: 1-65.
    Pubmed CrossRef
  16. Kashimura J. 2007. Sucrase activity inhibitor, glucoamylase activity inhibitor and food and feed containing the same. Global patent index - EP 1864670 A4.
  17. Khanahmadi M. 2004. Simulation and analysis of fluidized bed bioreactor for solid state fermentation. Sharif University of Technology, Iran.
  18. König J, Grasser R, Pikor H, Vogel K. 2002. Determination of xylanase, β-glucanase, and cellulase activity. Anal. Bioanal. Chem. 374: 80-87.
    Pubmed CrossRef
  19. Krishna SH, Srinivas ND, Raghavarao KSMS, Karanth NG. 2002. Reverse micellar extraction for downstream processing of proteins/enzymes. Adv. Biochem. Eng. Biotechnol. 75: 119-176.
    Pubmed CrossRef
  20. Liu JG, Xing JM, Chang TS, Liu HZ. 2006. Purification of nattokinase by reverse micelles extraction from fermentation broth: effect of temperature and phase volume ratio. Bioprocess Biosyst. Eng. 28: 267-273.
    Pubmed CrossRef
  21. Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ. 1951. Protein measurement with the Folin-phenol reagent. J. Biol. Chem. 193: 265-275.
    Pubmed
  22. Mahajan PB, Kolhekar SR, Borkar PS. 1983. Purification of amyloglucosidase. Anal. Biochem. 133: 482-485.
    CrossRef
  23. Mathew DS, Juang RS. 2007. Role of alcohols in the formation of inverse microemulsions and back extraction of proteins/enzymes in a reverse micellar system. Sep. Purif. Technol. 53: 199-215.
    CrossRef
  24. Nandini KE, Rastogi NK. 2010. Single step purification of lactoperoxidase from whey involving reverse micellesassisted extraction and its comparison with reverse micellar extraction. Biotechnol. Progress 26: 763-771.
    Pubmed CrossRef
  25. Nguyen QD, Rezessy-Szabo JM, Claeyssens M, Stals I, Hoschke A. 2002. Purification and characterisation of amylolytic enzymes from thermophilic fungus Thermomyces lanuginosus strain ATCC 34626. Enzyme Microb. Technol. 31: 345-352.
    CrossRef
  26. Noh KH, Imm JY. 2005. One-step separation of lysozyme by reverse micelles formed by the cationic surfactant, cetyldimethylammonium bromide. Food Chem. 93: 95-101.
    CrossRef
  27. Noh KH, Rhee MS, Imm JY. 2005. Separation of lactoferrin from model whey protein mixture by reverse micelles formed by cationic surfactant. Food Sci. Biotechnol. 14: 131-136.
  28. Ono K, Shigeta S, Oka S. 1988. Effective purification of glucoamylase in koji: a solid culture of Aspergillus oryzae on steamed rice by affinity chromatography using an immobilized acarbose (Bay g-5421). Agric. Biol. Chem. 52:1707-1714.
    CrossRef
  29. Paszczynski A, Miedziak I, Lobarzewski J, Kochmanska J, Trojanowski J. 1982. A simple method of affinity chromatography for the purification of glucoamylase obtained from Aspergillus niger C. FEBS Lett. 149: 63-66.
    CrossRef
  30. Pires MJ, Aires-Barros MR, Cabral JMS. 1996. Liquid-liquid extraction of proteins with reversed micelles. Biotechnol. Progress 12: 290-301.
    CrossRef
  31. Ramadas M, Holst O, Mattiasson B. 1995. Extraction and purification of amyloglucosidase produced by solid state fermentation with Aspergillus niger. Biotechnol. Tech. 9: 901-906.
    CrossRef
  32. Rodrigues EMG, Milagres AMF, Pessoa Jr A. 1999. Xylanase recovery: effect of extraction conditions on the AOT-reversed micellar systems using experimental design. Process Biochem. 34: 121-125.
    CrossRef
  33. Su CK, Chiang BH. 2003. Extraction of immunoglobulin-G from colostral whey by reverse micelles. J. D airy S ci. 86:1639-1645.
    CrossRef
  34. Tanuja S, Raghavarao KSMS, Gowthaman MK. 1997. Aqueous two phase extraction for downstream processing of amyloglucosidase. Process Biochem. 32: 635-641.
    CrossRef
  35. Tanuja S, Srinivas ND, Gowthman MK, Raghavarao KSMS. 2000. Aqueous two-phase extraction coupled with ultrafiltration for purification of amyloglucosidase. Bioprocess Eng. 23: 63-68.
    CrossRef
  36. Teotia S, Lata R, Khare SK, Gupta MN. 2001. One step purification of glucoamylase by affinity precipitation with alginate. J. Mol. Recogn. 14: 295-299.
    Pubmed CrossRef
  37. Yasuda M, Kuwae M, Matsushita H. 1989. Purification and properties of two forms of glucoamylase from Monascus sp. Agric. Biol. Chem. 53: 247-249.
    CrossRef
  38. Zivorad RL. 2004. Design of Experiment in Chemical Engineering. Wiley-VCH, Weinheim.

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Article

Research article

J. Microbiol. Biotechnol. 2014; 24(12): 1690-1698

Published online December 28, 2014 https://doi.org/10.4014/jmb.1407.07016

Copyright © The Korean Society for Microbiology and Biotechnology.

Reverse Micellar Extraction of Fungal Glucoamylase Produced in Solid-State Fermentation Culture

Aliakbar Paraj 1*, Morteza Khanahmadi 2, Keikhosro Karimi 1 and Mohammad J. Taherzadeh 3

1Department of Chemical Engineering, Isfahan University of Technology, Isfahan 84156-83111, Iran, 2Department of Agricultural Engineering, Isfahan Center for the Research of Agricultural Science & Natural Resources, Isfahan 81785-199, Iran, 3School of Engineering, University of Borås, Borås SE-501 90, Sweden

Received: July 8, 2014; Accepted: August 9, 2014

Abstract

Partial purification of glucoamylase from solid-state fermentation culture was, firstly,
investigated by reverse micellar extraction (RME). To avoid back extraction problems, the
glucoamylase was kept in the original aqueous phase, while the other undesired proteins/
enzymes were moved to the reverse micellar organic phase. The individual and interaction
effects of main factors (i.e., pH and NaCl concentration in the aqueous phase, and
concentration of sodium bis-2-ethyl-hexyl-sulfosuccinate (AOT) in the organic phase) were
studied using response surface methodology. The optimum conditions for the maximum
recovery of the enzyme were pH 2.75, 100 mM NaCl, and 200 mM AOT. Furthermore, the
optimum organic to aqueous volume ratio (Vorg/Vaq) and appropriate number of sequential
extraction stages were 2 and 3, respectively. Finally, 60% of the undesired enzymes including
proteases and xylanases were removed from the aqueous phase, while 140% of glucoamylase
activity was recovered in the aqueous phase and the purification factor of glucoamylase was
found to be 3.0-fold.

Keywords: Glucoamylase, Solid-State Fermentation, Purification, Reverse Micelle, AOT, Protein

References

  1. Andrews BA, Pyle DL, Asenjo JA. 1994. The effects of pH and ionic strength on the partitioning of four proteins in reverse micelle systems. Biotechnol. Bioeng. 43: 1052-1058.
    Pubmed CrossRef
  2. Beg QK, Kapoor M, Mahajan L, Hoondal GS. 2001. Microbial xylanases and their industrial applications: a review. Appl. Microbiol. Biotechnol. 56: 326-338.
    Pubmed CrossRef
  3. Bera MB, Panesar PS, Panesar R, Singh B. 2008. Application of reverse micelle extraction process for amylase recovery using response surface methodology. Bioprocess Biosyst. Eng. 31: 379-384.
    Pubmed CrossRef
  4. Bhatti HN, Rashid MH, Nawaz R, Asgher MM, Perveen R, Jabbar A. 2007. Purication and characterization of a novel glucoamylase from Fusarium solani. Food Chem. 103: 338-343.
    CrossRef
  5. Chen YL, Su CK, Chiang BH. 2006. Optimization of reversed micellar extraction of chitosanase produced by Bacillus cereus. Process Biochem. 41: 752-758.
    CrossRef
  6. Cortez EV Jr AP, Felipe MGA, Roberto IC, Vitolo M. 2004. Optimized extraction by cetyltrimethyl ammonium bromide reversed micelles of xylose reductase and xylitol dehydrogenase from Candida guilliermondii homogenate. J. Chromatogr. B 807: 47-54.
    Pubmed CrossRef
  7. Dean A, Voss D. 1999. Design and Analysis of Experiments. Springer-Verlag New York.
    CrossRef
  8. Ferreira LFP, Taqueda ME, Vitoloc M, Converti A, Pessoa Jr A. 2005. Liquid–liquid extraction of commercial glucose oxidase by reversed micelles. J. Biotechnol. 116: 411-416.
    Pubmed CrossRef
  9. George DA, Stuckey DC. 2010. Extraction of monoclonal antibodies (IgG1) using anionic and anionic/nonionic reverse micelles. Biotechnol. Progress 26: 1352-1360.
    Pubmed CrossRef
  10. Ghildyal NP, Ramakrishna M, Lonsane BK, Karanth NG. 1991. Efficient and simple extraction of mouldy bran in a pulsed column extractor for recovery of amyloglucosidase in concentrated form. Process Biochem. 26: 235-241.
    CrossRef
  11. Hasmann FA, Gurpilhares DB, Roberto IC, Pessoa Jr A. 2007. Response surface methodology for the evaluation of glucose-6-phosphate dehydrogenase enrichment process by soybean lecithin reversed micelles. J. Chromatogr. B 847: 262-266.
    Pubmed CrossRef
  12. Hayes DG, Marchio C. 1998. Expulsion of proteins from water-in-oil microemulsion by treatment with cosurfactant. Biotechnol. Bioeng. 59: 557-566.
    CrossRef
  13. Hemavathi AB, Hebbar HU, Raghavarao KSMS. 2010. Mixed reverse micellar systems for extraction and purification of βglucosidase. Sep. Purif. Technol. 71: 263-268.
    CrossRef
  14. Ikasari L, Mitchell DA. 1996. Leaching and characterization of Rhizopus oligosporus acid protease from solid-state fermentation. Enzyme Microb. Technol. 19: 171-175.
    CrossRef
  15. Kalisz HM. 1988. Microbial proteinases. Adv. Biochem. Eng. Biotechnol. 36: 1-65.
    Pubmed CrossRef
  16. Kashimura J. 2007. Sucrase activity inhibitor, glucoamylase activity inhibitor and food and feed containing the same. Global patent index - EP 1864670 A4.
  17. Khanahmadi M. 2004. Simulation and analysis of fluidized bed bioreactor for solid state fermentation. Sharif University of Technology, Iran.
  18. König J, Grasser R, Pikor H, Vogel K. 2002. Determination of xylanase, β-glucanase, and cellulase activity. Anal. Bioanal. Chem. 374: 80-87.
    Pubmed CrossRef
  19. Krishna SH, Srinivas ND, Raghavarao KSMS, Karanth NG. 2002. Reverse micellar extraction for downstream processing of proteins/enzymes. Adv. Biochem. Eng. Biotechnol. 75: 119-176.
    Pubmed CrossRef
  20. Liu JG, Xing JM, Chang TS, Liu HZ. 2006. Purification of nattokinase by reverse micelles extraction from fermentation broth: effect of temperature and phase volume ratio. Bioprocess Biosyst. Eng. 28: 267-273.
    Pubmed CrossRef
  21. Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ. 1951. Protein measurement with the Folin-phenol reagent. J. Biol. Chem. 193: 265-275.
    Pubmed
  22. Mahajan PB, Kolhekar SR, Borkar PS. 1983. Purification of amyloglucosidase. Anal. Biochem. 133: 482-485.
    CrossRef
  23. Mathew DS, Juang RS. 2007. Role of alcohols in the formation of inverse microemulsions and back extraction of proteins/enzymes in a reverse micellar system. Sep. Purif. Technol. 53: 199-215.
    CrossRef
  24. Nandini KE, Rastogi NK. 2010. Single step purification of lactoperoxidase from whey involving reverse micellesassisted extraction and its comparison with reverse micellar extraction. Biotechnol. Progress 26: 763-771.
    Pubmed CrossRef
  25. Nguyen QD, Rezessy-Szabo JM, Claeyssens M, Stals I, Hoschke A. 2002. Purification and characterisation of amylolytic enzymes from thermophilic fungus Thermomyces lanuginosus strain ATCC 34626. Enzyme Microb. Technol. 31: 345-352.
    CrossRef
  26. Noh KH, Imm JY. 2005. One-step separation of lysozyme by reverse micelles formed by the cationic surfactant, cetyldimethylammonium bromide. Food Chem. 93: 95-101.
    CrossRef
  27. Noh KH, Rhee MS, Imm JY. 2005. Separation of lactoferrin from model whey protein mixture by reverse micelles formed by cationic surfactant. Food Sci. Biotechnol. 14: 131-136.
  28. Ono K, Shigeta S, Oka S. 1988. Effective purification of glucoamylase in koji: a solid culture of Aspergillus oryzae on steamed rice by affinity chromatography using an immobilized acarbose (Bay g-5421). Agric. Biol. Chem. 52:1707-1714.
    CrossRef
  29. Paszczynski A, Miedziak I, Lobarzewski J, Kochmanska J, Trojanowski J. 1982. A simple method of affinity chromatography for the purification of glucoamylase obtained from Aspergillus niger C. FEBS Lett. 149: 63-66.
    CrossRef
  30. Pires MJ, Aires-Barros MR, Cabral JMS. 1996. Liquid-liquid extraction of proteins with reversed micelles. Biotechnol. Progress 12: 290-301.
    CrossRef
  31. Ramadas M, Holst O, Mattiasson B. 1995. Extraction and purification of amyloglucosidase produced by solid state fermentation with Aspergillus niger. Biotechnol. Tech. 9: 901-906.
    CrossRef
  32. Rodrigues EMG, Milagres AMF, Pessoa Jr A. 1999. Xylanase recovery: effect of extraction conditions on the AOT-reversed micellar systems using experimental design. Process Biochem. 34: 121-125.
    CrossRef
  33. Su CK, Chiang BH. 2003. Extraction of immunoglobulin-G from colostral whey by reverse micelles. J. D airy S ci. 86:1639-1645.
    CrossRef
  34. Tanuja S, Raghavarao KSMS, Gowthaman MK. 1997. Aqueous two phase extraction for downstream processing of amyloglucosidase. Process Biochem. 32: 635-641.
    CrossRef
  35. Tanuja S, Srinivas ND, Gowthman MK, Raghavarao KSMS. 2000. Aqueous two-phase extraction coupled with ultrafiltration for purification of amyloglucosidase. Bioprocess Eng. 23: 63-68.
    CrossRef
  36. Teotia S, Lata R, Khare SK, Gupta MN. 2001. One step purification of glucoamylase by affinity precipitation with alginate. J. Mol. Recogn. 14: 295-299.
    Pubmed CrossRef
  37. Yasuda M, Kuwae M, Matsushita H. 1989. Purification and properties of two forms of glucoamylase from Monascus sp. Agric. Biol. Chem. 53: 247-249.
    CrossRef
  38. Zivorad RL. 2004. Design of Experiment in Chemical Engineering. Wiley-VCH, Weinheim.