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Enhanced fructooligosaccharides and inulinase production by a Xanthomonas campestris pv. phaseoli KM 24 mutant

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Abstract

Xanthomonas campestris pv phaseoli produced an extracellular endoinulinase (9.24 ± 0.03 U mL−1) in an optimized medium comprising of 3% sucrose and 2.5% tryptone. X. campestris pv. phaseoli was further subjected to ethylmethanesulfonate mutagenesis and the resulting mutant, X. campestris pv. phaseoli KM 24 demonstrated inulinase production of 22.09 ± 0.03 U mL−1 after 18 h, which was 2.4-fold higher than that of the wild type. Inulinase production by this mutant was scaled up using sucrose as a carbon source in a 5-L fermenter yielding maximum volumetric (21,865 U L−1 h−1) and specific (119,025 U g−1 h−1) productivities of inulinase after 18 h with an inulinase/invertase ratio of 2.6. A maximum FOS production of 11.9 g L−1 h−1 and specific productivity of 72 g g−1 h−1 FOS from inulin were observed in a fermenter, when the mutant was grown on medium containing 3% inulin and 2.5% tryptone. The detection of mono- and oligosaccharides in inulin hydrolysates by TLC analysis indicated the presence of an endoinulinase. This mutant has potential for large-scale production of inulinase and fructooligosaccharides.

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References

  1. Kaur N, Gupta AK (2002) Application of inulin and oligofructose in health and nutrition. J Biosci 27:703–714

    Article  CAS  Google Scholar 

  2. Mazutti M, Bender JP, Treichel H, Luccio MD (2005) Optimization of inulinase production by solid-state fermentation using sugarcane bagasse as substrate. Enzyme Microb Technol 39:56–59

    Article  CAS  Google Scholar 

  3. Van Loo J, Coussement P, De Leenheer L, Hoebregs H, Smits G (1995) On the presence of inulin and oligofructose as natural ingredients in western diet. CRC Crit Rev Food Sci Nut 35:525–552

    Article  Google Scholar 

  4. Yun JW (1996) Fructooligosaccharides—occurrence, preparation, and application. Enzyme Microb Technol 19:107–117

    Article  CAS  Google Scholar 

  5. Vandamme EJ, Deryeke DG (1983) Microbial inulinases: fermentation process, properties and applications. Advan Appl Microbiol 29:139–176

    Article  CAS  Google Scholar 

  6. Singh P, Gill PK (2006) Production of inulinases: recent advances. Food Technol Biotechnol 44:151–162

    CAS  Google Scholar 

  7. Cho YJ, Sinha J, Park JP, Yun JW (2001) Production of inulooligosaccharides from chicory extract by endoinulinase from Xanthomonas oryzae No. 5. Enzyme Microb Technol 28:439–445

    Article  CAS  Google Scholar 

  8. Nakamura T, Nagatomo Y, Hamada S, Nishino Y, Ohta K (1994) Occurrence of two forms of extracellular endoinulinase from Aspergillus niger mutant 817. J Ferment Bioeng 78:134–139

    Article  CAS  Google Scholar 

  9. Sharma AD, Gill PK, Bhullar SS, Singh P (2005) Improvement in inulinase production by simultaneous action of physical and chemical mutagenesis in Penicillium purpurogenum. World J Microbiol Biotechnol 21:929–932

    Article  CAS  Google Scholar 

  10. Kamal FHM, Mahnaz MA, Seyed AM (2003) Mutagenesis of Xanthomonas campestris and selection of strains with enhanced xanthan production. Iran Biomed J 7:91–98

    CAS  Google Scholar 

  11. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428

    Article  CAS  Google Scholar 

  12. Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356

    Article  CAS  Google Scholar 

  13. Park JP, Yun JW (2001) Utilization of chicory roots for microbial endoinulinase production. Lett Appl Microbiol 33:183–187

    Article  CAS  Google Scholar 

  14. Gill PK, Sharma AD, Harchand RK, Singh P (2003) Effects of media supplements and culture conditions on inulinase production by an actinomycete strain. Bioresour Technol 87:359–362

    Article  CAS  Google Scholar 

  15. Cruz VDA, Belie JG, Belline MZ, Cruz R (1998) Production and action pattern of inulinase from Aspergillus niger-245: hydrolysis of inulin from several sources. Revista de Microbiologia 29:301–306

    Article  CAS  Google Scholar 

  16. Dorta C, Cruz R, Oliva-Neto P, José D, Moura C (2006) Sugarcane molasses and yeast powder used in the fructooligosaccharides production by Aspergillus japonicus- FCL 119T and Aspergillus niger ATCC 20611. J Ind Microbiol Biotechnol 33:1003–1009

    Article  CAS  Google Scholar 

  17. Gong F, Sheng J, Chi Z, Li J (2007) Inulinase production by a marine yeast Pichia guilliermondii and inulin hydrolysis by the crude inulinase. J Ind Microbiol Biotechnol 34:179–185

    Article  CAS  Google Scholar 

  18. Elyachioui M, Horeiz JP, Taillez R (1992) General properties of extracellular bacterial inulinase. J Appl Bacteriol 73:514–519

    CAS  Google Scholar 

  19. Park JP, Bae JT, You DJ, Kim BW, Yun JW (1999) Production of inulooligosaccharides from inulin by a novel endoinulinase from Xanthomonas sp. Biotechnol Lett 21:1043–1046

    Article  CAS  Google Scholar 

  20. Vishwanathan P, Kulkarni PR (1995) Enhancement of inulinase production by Aspergillus niger van Teighem. J Appl Bacteriol 78:384–386

    Google Scholar 

  21. Ayyachamy M, Khelawan K, Pillay D, Permaul K, Singh S (2007) Production of inulinase by Xanthomonas campestris pv phaseoli using onion (Allium cepa) and garlic (Allium sativum) peels in solid state cultivation. Lett Appl Microbiol 45:439–444

    Article  CAS  Google Scholar 

  22. Cho YJ, Yun JW (2002) Purification and characterization of endoinulinase from Xanthomonas oryzae No. 5. Process Biochem 37:1325–1331

    Article  CAS  Google Scholar 

  23. Ettalibi M, Baratti JC (1987) Purification, properties and comparison of invertase, exoinulinases and endoinulinases of Aspergillus ficuum. Appl Microbiol Biotechnol 26:13–20

    Article  CAS  Google Scholar 

  24. Silva-santisteban BOY, Filho FM (2005) Agitation, aeration and shear stress as key factors in inulinase production by Kluyveromyces marxianus. Enzyme Microb Technol 36:717–724

    Article  CAS  Google Scholar 

  25. Nguyen QD, Mattes F, Hoschke AG, Rezessy-Szabo J, Bhat MK (1999) Production, purification and identification of fructooligosaccharides produced by β fructofuranosidase from Aspergillus niger IMI 303386. Biotechnol Lett 21:183–186

    Article  CAS  Google Scholar 

  26. Park JP, Kim DH, Kim DS, Yun JW (1998) Enzymatic production of inulooligosaccharides from chicory juice. Biotechnol Lett 20:385–388

    Article  CAS  Google Scholar 

  27. Ohta K, Suetsugu N, Nakamura T (2002) Purification and properties of an extracellular inulinase from Rhizopus sp. strain TN-96. J Biosci Bioeng 94:78–80

    Article  CAS  Google Scholar 

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Acknowledgments

This study was supported by grants from the National Research Foundation, Republic of South Africa. The authors are thankful to Prof. Bernard Prior, University of Stellenbosch, South Africa, for his valuable suggestions and critical evaluation of the manuscript.

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Correspondence to Manimaran Ayyachamy.

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Naidoo, K., Ayyachamy, M., Permaul, K. et al. Enhanced fructooligosaccharides and inulinase production by a Xanthomonas campestris pv. phaseoli KM 24 mutant. Bioprocess Biosyst Eng 32, 689–695 (2009). https://doi.org/10.1007/s00449-008-0293-6

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  • DOI: https://doi.org/10.1007/s00449-008-0293-6

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