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Oxygen supply in Bacillus thuringiensis fermentations: bringing new insights on their impact on sporulation and δ-endotoxin production

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Abstract

The growth kinetics, sporulation, and toxicity of Bacillus thuringiensis var. israelensis were evaluated through the analysis of batch cultures with different dissolved oxygen (DO) profiles. Firstly, DO was maintained constant at 5%, 20%, or 50% throughout fermentation in order to identify the most suitable one to improve the main process parameters. Higher biomass concentration, cell productivity, and cell yield based on glucose were obtained with 50% DO. The higher aeration level also resulted in higher spore counts and markedly improved the toxic activity of the fermentation broth, which was 9-fold greater than that obtained with 5% DO (LC50 of 39 and 329 mg/L, respectively). Subsequently, using a two-stage oxygen supply strategy, DO was kept at 50% during the vegetative and transition phases until the maximum cell concentration was achieved. Then, DO was changed to 0%, 5%, 20%, or 100% throughout sporulation and cell lysis phases. The interruption of oxygen supply strongly reduced the spore production and thoroughly repressed the toxin synthesis. On the contrary, when DO was raised to 100% of saturation, toxic activity increased approximately four times (LC50 of 8.2 mg/L) in comparison with the mean values reached with lower DO levels, even though spore counts were lower than that from the 50% DO assay. When pure oxygen was used instead of normal air, it was possible to obtain 70% of the total biomass concentration achieved in the air assays; however, cultures did not sporulate and the toxin synthesis was consequently suppressed.

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Acknowledgments

The authors gratefully acknowledge the financial support of FAPERJ (Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro, Brazil) and Dr. Ines C. Roberto for her support with fermentation facilities.

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Authors and Affiliations

  1. Laboratório de Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, CEP 28013-602, Campos dos Goytacazes, RJ, Brazil

    Fabrízio Siqueira Boniolo, Raphael Cardoso Rodrigues, Tânia Jacinto & Marília Amorim Berbert-Molina

  2. Departamento de Biotecnologia, Escola de Engenharia de Lorena, Universidade de São Paulo, CEP 12602-810, Lorena, SP, Brazil

    Arnaldo Márcio Ramalho Prata

  3. Laboratório de Biologia Celular e Tecidual, Universidade Estadual do Norte Fluminense Darcy Ribeiro, CEP 28013-602, Campos dos Goytacazes, RJ, Brazil

    Maria Luisa López

  4. Instituto de Biotecnologia, Universidade de Caxias do Sul, CEP 95001-970, Caxias do Sul, RS, Brazil

    Mauricio Moura da Silveira

Authors
  1. Fabrízio Siqueira Boniolo
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  2. Raphael Cardoso Rodrigues
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  3. Arnaldo Márcio Ramalho Prata
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  4. Maria Luisa López
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  5. Tânia Jacinto
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  6. Mauricio Moura da Silveira
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  7. Marília Amorim Berbert-Molina
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Correspondence to Marília Amorim Berbert-Molina.

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Boniolo, F.S., Rodrigues, R.C., Prata, A.M.R. et al. Oxygen supply in Bacillus thuringiensis fermentations: bringing new insights on their impact on sporulation and δ-endotoxin production. Appl Microbiol Biotechnol 94, 625–636 (2012). https://doi.org/10.1007/s00253-011-3746-9

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  • DOI: https://doi.org/10.1007/s00253-011-3746-9

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