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Optimization of Pseudoalteromonas sp. JYBCL 1 culture conditions, medium composition and extracellular β-agarase activity

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Abstract

Extracellular agarase produced by the Pseudoalteromonas strain JYBCL 1 is used in a variety of applications in the biotechnology, pharmaceutical, cosmetic, and food industries. The optimization of culture conditions for agarase-producing microbes and agarase activity is thus an important consideration in many industrial applications. In this study, the optimum medium composition and culture conditions for the JYBCL 1 strain were determined using the “one factor at a time” (OFAT) method and a Plackett-Burman design. Optimal cell growth was obtained at a temperature of 25°C and when 10 g/L tryptone was present in the culture medium. Optimal agarase activity occurred at a temperature of 40°C and at pH 6. The presence of carbonyl groups in the extracellular agarase hydrolysis products was verified using FT-IR. LC-MS identified the hydrolyzates as neoagarohexaose, neoagarotetraose, and neoagarobiose. The extracellular agarase produced by the JYBCL 1 strain used in this study was identified as β-agarase by 13C-NMR spectroscopy.

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References

  1. Ohta, Y., Y. Hatada, M. Miyazaki, Y. Nogi, S. Ito, and K. Horikoshi (2005) Purification and characterization of a novel α-agarase from a Thalassomonas sp. Curr. Microbiol. 50: 212–216.

    Article  CAS  Google Scholar 

  2. Potin, P., C. Richard, C. Rochas, and B. Kloareg (1993) Purification and characterization of the alpha-agarase from Alteromonas agarlyticus (Cataldi) comb. nov., strain GJ1B. Eur. J. Biochem. 214: 599–607.

    Article  CAS  Google Scholar 

  3. Lakshmikanth, M., S. Manohar, Y. Souche, and J. Lalitha (2006) Extracellular β-agarase LSL-1 producing neoagarobiose from a newly isolated agar-liquefying soil bacterium, Acinetobacter sp., AG LSL-1. World J. Microbiol. Biotechnol. 22: 1087–1094.

    Article  CAS  Google Scholar 

  4. Van der Meulen, H. J. and W. Harder (1975) Production and characterization of the agarase of Cytophaga flevensis. Antonie. Van. Leeuwenhoek. 41: 431–447.

    Article  Google Scholar 

  5. Ha, J. C., G. T. Kim, S. K. Kim, T. K. Oh, J. H. Yu, and I. S. Kong (1997) Beta-agarase from Pseudomonas sp. W7: Purification of the recombinant enzyme from Escherichia coli and the effects of salt on its activity. Biotechnol. Appl. Biochem. 26: 1–6.

    CAS  Google Scholar 

  6. Belas, R. (1989) Sequence analysis of the agrA gene encoding β-agarase from Pseudomonas atlantica. J. Bacteriol. 171: 602–605.

    CAS  Google Scholar 

  7. Kang, N. Y., Y. L. Choi, Y. S. Cho, B. K. Kim, B. S. Jeon, J. Y. Cha, C. H. Kim, and Y. C. Lee (2003) Cloning, expression and characterization of a beta-agarase gene from a marine bacterium, Pseudomonas sp. SK38. Biotechnol. Lett. 25: 1165–1170.

    Article  CAS  Google Scholar 

  8. Morrice, L. M., M. W. McLean, W. F. Long, and F. B. Williamson (1983) Beta-agarases I and II from Pseudomonas atlantica. Substrate specificities. Eur. J. Biochem. 137: 149–154.

    Article  CAS  Google Scholar 

  9. Araki, T., M. Hayakawa, L. Zhang, S. Karita, and T. Morishita (1998) Purification and characterization of agarases from a marine bacterium, Vibrio sp. PO-303. J. Mar. Biotechnol. 6: 260–265.

    Google Scholar 

  10. Sugano, Y., I. Terada, M. Arita, and M. Noma (1993) Purification and characterization of a new agarase from a marine bacterium, Vibrio sp. strain JT0107. Appl. Environ. Microbiol. 59: 1549–1554.

    CAS  Google Scholar 

  11. Hu, Z., B. K. Lin, Y. Xu, M. Q. Zhong, and G. M. Liu (2008) Production and purification of agarase from a marine agarolytic bacterium Agarivorans sp. Z105. J. Appl. Microbiol. 106: 181–190.

    Article  Google Scholar 

  12. Ting, F. X., H. Lin, and S. M. Kim (2008) Purification and characterization of a novel β-agarase, AgaA34, from Agarivorans albus YKW-34. Appl. Microbiol. Biotechnol. 78: 265–273.

    Article  Google Scholar 

  13. Ting, F. X., C. H. Pan, H. Lin, and S. M. Kim (2009) Gene Cloning, Expression, and Characterization of a â-Agarase, AgaB34, from Agarivorans albus YKW-34. J. Microbiol. Biotechnol. 19: 257–264.

    Google Scholar 

  14. Kim, H. T., S. Y. Lee, D. H. Lee, H. S. Kim, W. G. Bang, K. H. Kim, and I. G. Choi (2010) Overexpression and molecular characterization of Aga50D from Saccharophagus degradans 2–40: An exo-type β-agarase producing neoagarobiose. Appl. Microbiol. Biotechnol. 86: 227–234.

    Article  CAS  Google Scholar 

  15. Ohta, Y., Y. Hatada, Y. Nogi, Z. Li, S. Ito, and K. Horikoshi (2004) Cloning, expression, and characterization of a glycoside hydrolase family 86 β-agarase from a deep-sea Microbulbifer-like isolate. Appl. Microbiol. Biotechnol. 66: 266–275.

    Article  CAS  Google Scholar 

  16. Ma, C., X. Lu, C. Shi, J. Li, Y. Gu, Y. Ma, Y. Chu, F. Han, Q. Gong, and W. Yu (2007) Molecular cloning and characterization of a novel betaagarase, AgaB, from marine Pseudoalteromonas sp. CY24. J. Biol. Chem. 282: 3747–3754.

    Article  CAS  Google Scholar 

  17. Chiura, H. and K. Tsukamoto (2000) Purification and characterisation of novel agarase secreted by marine bacterium, Pseudoalteromonas sp. Strain CKT1. Microbes. Environ. 15: 11–22.

    Article  Google Scholar 

  18. Schroeder, D. C., M. A. Jaffer, and V. E. Coyne (2003) Investigation of the role of a beta (1–4) agarase produced by Pseudoalteromonas gracilis B9 in eliciting disease symptoms in the red alga Gracilaria gracilis. Microbiol. 149: 2919–2929.

    Article  CAS  Google Scholar 

  19. Oh, C., C. Nikapitiya, Y. Lee, I. Whang, S. J. Kim, D. H. Kang, and J. Lee (2010) Cloning, purification and biochemical characterization of beta agarase from the marine bacterium Pseudoalteromonas sp. AG4. J. Ind. Biotechnol. 37: 483–494.

    Article  CAS  Google Scholar 

  20. Oh, C., C. Nikapitiya, Y. Lee, I. Whang, D. H. Kang, S. J. Heo, Y. U. Choi, and J. Lee (2010) Molecular cloning, characterization and enzymatic properties of a novel beta-agarase from a marine isolate Psudoalteromonas sp. AG52. B. J. Microbiol. 41: 876–889.

    Article  CAS  Google Scholar 

  21. Jam, M., D. Flament, J. Allouch, P. Potin, L. Thion, B. Kloareg, M. Czjzek, W. Helbert, G. Michel, and T. Barbeyron (2005) The endo-β-agarases AgaA and AgaB from the marine bacterium Zobellia galactanivorans: Two paralogue enzymes with different molecular organizations and catalytic behaviours. Biochem. J. 385: 703–713.

    Article  CAS  Google Scholar 

  22. Kirimura, K., N. Masuda, Y. Iwasaki, H. Nakagawa, R. Kobayashi, and S. Usami (1999) Purification and characterization of a novel β-agarase from an alkalophilic bacterium, Alteromonas sp. E-1. J. Biosci. Bioeng. 87: 436–441.

    Article  CAS  Google Scholar 

  23. Vera, J., R. Alvarez, E. Murano, J. C. Slebe, and O. Leon (1998) Identification of a marine agarolytic Pseudoalteromonas isolate and characterization of its extracellular agarase. Appl. Environ. Microbiol. 64: 4378–4383.

    CAS  Google Scholar 

  24. Rhee, Y. J., R. H. Cho, W. C. Kim, D. Y. Jun, I. K. Rhee, and Y. H. Kim (2010) Isolation of a novel freshwater agarolytic Cellvibrio sp. KY-YJ-3 and characterization of its extracellular â-Agarase. J. Microbiol. Biotechnol. 20: 1378–1385.

    Article  CAS  Google Scholar 

  25. Holmstrom, C. and S. Kjelleberg (1999) Marine Pseudoalteromonas species are associated with higher organisms and produce biologically active extracellular agents organisms and produce biologically active extracellular agents. FEMS. Microbiol. Ecol. 30: 285–293.

    Article  CAS  Google Scholar 

  26. Oh, Y. H., C. K. Jung, and J. W. Lee (2011) Isolation and characterization of a novel Agarase-producing Pseudoalteromonas spp. Bacterium from the guts of spiny turban shells. J. Microbiol. Biotechnol. 21: 818–821.

    Article  CAS  Google Scholar 

  27. Lakshmikanth, M., S. Manohar, J. Patnakar, P. Vaishampayan, Y. Shouche, and J. Lalitha (2006) Optimization of culture conditions for the production of extracellular agarases from newly isolated Pseudomonas aeruginosa AG LSL-11. World J. Microbiol. Biotechnol. 22: 531–537.

    Article  CAS  Google Scholar 

  28. Fu, X. T., H. Lin, and S. M. Kim (2009) Optimization of medium composition and culture conditions for agarase production by Agarivorans albus YKW-34. Proc. Biochem. 44: 1158–1163.

    Article  CAS  Google Scholar 

  29. Wang, J., X. Jiang, H. Mouand, and H. Guan (2004) Anti-oxidation of agarooligosaccharides produced by agarase from a marine bacterium. J. Appl. Phycol. 16: 333–340.

    Article  CAS  Google Scholar 

  30. Kobayashi, R., M. Takisada, T. Suzuki, K. Kirimura, and S. Usami (1997) Neoagarobiose as a novel moisturizer with whitening effect. Biosci. Biotech. Biochem. 61: 162–163.

    Article  CAS  Google Scholar 

  31. Wu, S. C., T. N. Wen, and C. L. Pan (2005) Algal-oligosaccharide-lysates prepared by two bacterial agarases stepwise hydrolyzed and their anti-oxidative properties. Fish. Sci. 71: 1149–1159.

    Article  CAS  Google Scholar 

  32. Fu, X. T. and S. M. Kim (2010) Agarase: Review of major sources, categories, purification method, enzyme characteristics and applications. Mar. Drugs. 8: 200–218.

    Article  CAS  Google Scholar 

  33. Choi, H. J., J. B. Hong, J. J. Park, W. J. Chi, M. C. Kim, Y. K. Chang, and S. K. Hong (2011) Production of agarase from a novel micrococcus sp. GNUM-08124 strain isolated from the East Sea of Korea. Biotechnol. Bioproc. Eng. 16: 81–88.

    Article  CAS  Google Scholar 

  34. Khambhaty, Y., K. Mody, and B. Jha (2008) Purification, characterization and application of a novel extracellular agarase from a marine bacillus megaterium. Biotechnol. Bioproc. Eng. 13: 584–591.

    Article  CAS  Google Scholar 

  35. Stuart, B. and D. J. Ando (1997) Biological applications of infrared spectroscopy. pp. 65–68. John Wiley & Sons, NY, USA.

    Google Scholar 

  36. Rochas, C., M. Lahaye, W. Yaphe, and M. T. P. Viet (1986) 13CN. M.R.—Spectroscopic investigation of agarose oligomers, Carbo. Res. 148: 199–207.

    Article  CAS  Google Scholar 

  37. Rochas, C., P. Potin, and B. Kloareg (1994) NMR spectroscopic investigation of agarose oligomers produced by an α-agarase. Carbo. Res. 253: 69–77.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, Sogang University, Seoul, 121-742, Korea

    Changkyou Jung, Hyuckjin Kwon & Jinwon Lee

  2. Department of Chemical Engineering, Kwangwoon University, Seoul, 139-701, Korea

    Chulhwan Park

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  1. Changkyou Jung
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Correspondence to Jinwon Lee.

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Jung, C., Kwon, H., Park, C. et al. Optimization of Pseudoalteromonas sp. JYBCL 1 culture conditions, medium composition and extracellular β-agarase activity. Biotechnol Bioproc E 17, 937–945 (2012). https://doi.org/10.1007/s12257-012-0009-2

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  • DOI: https://doi.org/10.1007/s12257-012-0009-2

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