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Phytase production by high cell density culture of recombinant Bacillus subtilis

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Abstract

Bacillus subtilis BD170, harboring a plasmid pGT44[phyC] carrying the phytase gene (phyC) and a phosphate-depletion inducible pst-promoter, was grown in a 2 l bioreactor. Using a controlled feeding of glucose, high cell densities of 32 and 56 g dry cell weight l−1 were achieved with peptone and yeast extract, respectively, as the complex nitrogen sources in a semi-defined growth medium. The fed-batch protocol was applied to production of recombinant phytase and a high extracellular phytase activity (48 U ml−1) was reached with peptone. Although the yeast extract feeding resulted in a higher cell density, it was unsuitable as a medium component for phytase expression due to its relatively high phosphate content.

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References

  • Cayuela C, Kai K, Park YS, Iijima S, Kobayashi T (1993) Insectiside production by recombinant Bacillus subtilis 1A96 in fedbatch culture with control of glucose concentration. J. Ferment. Bioeng. 75: 383-386.

    Google Scholar 

  • Doran PM (1997) Bioprocess Engineering Principles. London: Academic Press Ltd.

    Google Scholar 

  • Giesecke UE, Bierbaum G, Rudde H, Spohn U, Wandrey C (1991) Production of alkaline protease with Bacillus licheniformis in a controlled fed-batch process. Appl. Microbiol. Biotechnol. 35: 720-724.

    Google Scholar 

  • Kai K, Park YS, Iijima S, Kobayashi T (1992) High concentration of Bacillus subtilis spo-mutant carrying a recombinant plasmid. J. Ferment. Bioeng. 73: 248-250.

    Google Scholar 

  • Kang BC, Lee SY, Chang HN (1992) Enhanced spore production of Bacillus thuringiensis by fed-batch culture. Biotechnol. Lett. 14: 721-726.

    Google Scholar 

  • Kerovuo J, Lauraeus M, Nurminen P, Kalkkinen N, Apajalahti J (1998) Isolation, characterization, molecular gene cloning, and sequencing of a novel phytase from Bacillus subtilis. Appl. Environ. Microbiol. 64: 2079-2085.

    Google Scholar 

  • Kerovuo J, von Weymarn N, Povelainen M, Auer S, Miasnikov A (2000) A new efficient expression system for Bacillus and its ap766 plication to production of recombinant phytase. Biotechnol. Lett. 22: 1311-1317.

    Google Scholar 

  • Kim YO, Lee JK, Oh BC, Oh TK (1999) High-level expression of a recombinant thermostable phytase in Bacillus subtilis. Biosci. Biotechnol. Biochem. 63: 2205-2207.

    Google Scholar 

  • Lee AY (1996) High cell-density culture of Escherichia coli. Trends Biotechnol. 14: 98-105.

    Google Scholar 

  • Mignone CF, Rossa CA (1993) A simple method for designing fed-batch cultures with linear gradient feed of nutrients. Process Biochem. 28: 405-410.

    Google Scholar 

  • Nakano MM, Dailly YP, Zuber P, Clark DP (1997) Characterization on anaerobic fermentative growth of Bacillus subtilis: identification of fermentation end products and genes required for growth. J. Bacteriol. 179: 6749-6755.

    Google Scholar 

  • Park YS, Dohjima T, Okabe M (1996) Enhanced α-amylase production in recombinant Bacillus brevis by fed-batch culture with amino acid control. Biotechnol. Bioeng. 49: 36-44.

    Google Scholar 

  • Park YS, Kai K, Iijima S, Kobayashi T (1992) Enhanced β-galactosidase production by high cell-density culture of recombinant Bacillus subtilis with glucose concentration control. Biotechnol. Bioeng. 40: 686-696.

    Google Scholar 

  • Speck EL, Freese E (1973) Control of metabolite secretion in Bacillus subtilis. J. Gen. Microbiol. 78: 261-275.

    Google Scholar 

  • Yoon SM, Kim SC, Kim JH (1994) Identification of inhibitory metabolites in high density culture of recombinant Bacillus megaterium pCK108. Biotechnol. Lett. 16: 1011-1014.

    Google Scholar 

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Vuolanto, A., von Weymarn, N., Kerovuo, J. et al. Phytase production by high cell density culture of recombinant Bacillus subtilis. Biotechnology Letters 23, 761–766 (2001). https://doi.org/10.1023/A:1010369325558

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  • DOI: https://doi.org/10.1023/A:1010369325558

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