Abstract
A large amount of adenosine triphosphate with high energy phosphate bonds is required for uridine triphosphate regeneration during curdlan biosynthesis by Agrobacterium sp. ATCC 31749. To supply high energy for curdlan synthesis, three low-polyphosphates (Na4P2O7, Na5P3O10, and (NaPO3)6) with higher energy phosphate bonds were employed to substitute for KH2PO4-K2HPO4 in fermentation medium. Two genes encoding the polyphosphate metabolizing enzymes, polyphosphate kinase and exopolyphosphatase, were amplified and showed 95% homology to those in Agrobacterium sp. C58 by sequence analysis. The curdlan yields were enhanced by 23 and 134% when phosphate concentrations 0.024 mol/L of Na5P3O10 and 0.048 mol/L of (NaPO3)6 respectively, were added in the medium. The maximum curdlan yield of 30 ± 1.02 g/L was obtained with the addition of 0.048 mol/L of (NaPO3)6 with 5 g/L CaCO3 in the medium. When CaCO3 was removed from the culture and the three lowpolyphosphates were added, the pH and biomass yield dropped remarkably and little or no curdlan was produced. The culture containing 0.048 mol/L of (NaPO3)6 was mixed with KH2PO4-K2HPO4 and CaCO3 in the medium, but showed no effect on curdlan production. However, curdlan yield was improved by 49 ∼ 60% when CaCO3 was removed from the medium and KH2PO4-K2HPO4 acted as a buffer. It appears that the positive effect of (NaPO3)6 on curdlan production required the buffering capacity of CaCO3 and the absence of KH2PO4-K2HPO4 competing as a phosphate supplier.
Similar content being viewed by others
References
Harada, T., K. Fujimori, S. Hirose, and M. Masada (1966) Growth and glucan (10C3K) production by a mutant of Alcaligenes faecalis vax. myxogenes in defined medium. Agric. Biol. Chem. 30: 764–769.
Jagodzinski, P. P., R. Wiaderkiewicz, G. Kurzawski, M. Kloczewiak, and H. Nakashima (1994) Mechanism of the inhibitory effect of curdlan sulfate on HIV-I infection in vitro. Virology 202: 735–745.
Mikio, K., O. Yoshiro, O. Hirotomo, K. Yoshikazu, I. Hiroshi, M. Hisashi, and K. Takeshi (1995) Water soluble β-(1,3)-glucan derivative and antiviral agent containing the derivative. Jpn Patent 07228601.
Evans, S. G., D. Morrison, Y. Kaneko, and I. Havlik (1998) The effect of curdlan sulfate on development in vitro of Plasmodium falciparum. Trans. R. Soc. Trop. Med. Hyg. 92: 87–89.
Sutherland, I. W. and D. C. Elwood (1979) Microbial exopolysaccharide-industrial polymers of current and future potentia1. pp. 422–427. In: Bull A. T. (ed.). Microbial Technology: Current States and Future Prospects. Cambridge University Press, London.
Lawford, H. G., K. R. Phillips, and G. R. Lawford (1982) Two stage continuous process for the production of thermogelable curdlan-type exopolysaccharide. Biotechnol. Lett. 4: 689–694.
Lee, I. Y., M. K. Kim, J. H. Lee, W. T. Seo, J. K. Jung, H. W. Lee, and Y. H. Park (1999) Influence of agitation speed on production of curdlan by Agrobacterium species. Bioproc. Eng. 20: 283–287.
Kim, M. K., I. Y. Lee, J. H. Lee, K. T. Kim, Y. H. Rhee, and Y. H. Park (2000) Residual phosphate concentration under nitrogen 1imiting conditions regulates curdlan production in Agrobacterium species. J. Ind. Microbiol. Biotechnol. 25: 180–183.
Lee, J. H. and I. Y. Lee (2001) Optimization of uracil addition for curdlan (β-1,3-glucan) production by Agrobacterium sp. Biotechnol. Lett. 23: 1131–1134.
Lee, J. H., I. Y. Lee, M. K. Kim, and Y. H. Park (1999) Optimal pH control of batch process for production of curdlan by Agrobacterium species. J. Ind. Microbiol. Biotechnol. 23: 143–148.
Wu, J., X. Zhan, H. Liu, and Z. Zheng (2008) Enhanced production of curdlan by Alcaligenes faecalis by selective feeding with ammonia water during the cell growth phase of fermentation. Chin. J. Biotech. 24: 1035–1039.
Kim, M. K., I. Y. Lee, J. H. Ko, Y. H. Rhee, and Y. H. Park (1999) Higher intracellular levels of uridine monophosphate under nitrogen 1imited conditions enhance metabolic flux of curdlan synthesis in Agrobacterium species. Biotechnol. Bioeng. 62: 317–323.
West, T. P. (2006) Pyrimidine base supplementation effects curdlan production in Agrobacterium sp. ATCC 31749. J. Basic. Microbiol. 46: 153–157.
Gummadi, S. N. and K. Kumar (2005) Production of extracellular water insoluble β-1,3-glucan (Curdlan) from Bacillus sp. SNC07. Biotechnol. Bioproc. Eng. 10: 546–551.
Zheng, Z. Y., J. W. Lee, X. B. Zhan, Z. P. Shi, L. Wang, L. Zhu, J. R. Wu, and C. C. Lin (2007) Effect of metabolic structures and energy requirements on curdlan production by Alcaligenes faecalis. Biotechnol. Bioproc. Eng. 12: 359–365.
Liao, X., T. Deng, Y. Zhu, G. Du, and J. Chen (2008) Enhancement of glutathione production by altering adenosine metabolism of Escherichia coli in a coupled ATP regeneration system with Saccharomyces cerevisiae. J. Appl. Microbiol. 104: 345–352.
Noguchi, T. (1998) Use of Escherichia coli polyphosphate kinase for oligosaccharide synthesis. Biosci. Biotechnol. Biochem. 62: 1594–1596.
Ahn, K. and A. Kornberg (1990) Polyphosphate kinase from Escherichia coli: Purification and demonstration of a phosphoenzyme intermediate. J. Biol. Chem. 265: 11734–11739.
Akiyama, M., E. Crooke, and A. Kornberg (1993) An exopolyphosphatase of Escherichia coli: the enzyme and its ppx gene in a polyphosphate operon. J. Biol. Chem. 268: 633–639.
Murata, K., T. Uchida, J. Kato, and I. Chibata (1988) Polyphosphate kinase: Distribution, some properties and its application as an ATP regeneration system. Agric. Biol. Chem. 52: 1471–1477.
Kulaev, I. and V. Vagabov (1999) New aspects of inorganic polyphosphate metabolism and function. J. Biosci. Bioeng. 88: 111–129.
Kim, K. S., N. N. Rao, and D. Cresson (2002) Inorganic polyphosphate is essential for long-term survival and virulence factors in Shigella and Salmonella spp. Proc. Natl. Acad. Sci. 99: 7675–7680.
Werner, T. P., N. Amrhein, and F. M. Freimoser (2007) Inorganic polyphosphate occurs in the cell wall of Chlamydomonas reinhardtii and accumulates during cytokinesis. Plant Biol. 7: 1–11.
Nocek, B., S. Kochinyan, M. Proudfoot, G. Brown, E. Evdokimova, J. Osipiuk, A. M. Edwards, A. Savchenko, A. Joachimiak, and A. F. Yakunin (2008) Polyphosphate-dependent synthesis of ATP and ADP by the family-2 polyphosphate kinases in bacteria. Proc. Natl. Acad. Sci. 105: 17730–17735.
Iwamoto, S., K. Motomura, Y. Shinoda, M. Urata, J. Kato, N. Takiguchi, H. Ohtake, R. Hirota, and A. Kuroda (2007) Use of an Escherichia coli recombinant producing thermostable polyphosphate kinase as an ATP regenerator to produce fructose 1,6-diphosphate. Appl. Environ. Microbiol. 73: 5676–5678.
Miller, G. L. (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 31: 426–428.
Scroggins, L. H. (1968) Spectrophotometric microchemical phosphorus determination: A quantitative oxygen flask procedure applicable to problem organophosphorus compounds. Microchem. J. 13: 385–391.
Jin, L. H., H. J. Um, C. J. Yin, Y. H. Kim, and J.H. Lee (2008) Proteomic analysis of curdlan-producing Agrobacterium sp. in response to pH downshift. J. Biotechnol. 138: 80–87.
Cristian, V., M. Cecilia, P. Alberto, A. Juan, J. Carlos, and C. Francisco (2010) New structural and functional defects in polyphosphate deficient bacteria: A cellular and proteomic study. BMC Microbiol. 10: 1–7.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yu, L., Wu, J., Liu, J. et al. Enhanced curdlan production in Agrobacterium sp. ATCC 31749 by addition of low-polyphosphates. Biotechnol Bioproc E 16, 34–41 (2011). https://doi.org/10.1007/s12257-010-0145-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12257-010-0145-5