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Selection and Optimization of Bacillus atrophaeus Inoculum Medium and its Effect on Spore Yield and Thermal Resistance

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Abstract

Bacillus atrophaeus’s spores are used as biological indicators to monitor sterilization processes and as a Bacillus anthracis surrogate in the development and validation of biosafety methods. The regular use of biological indicators to evaluate the efficiency of sterilization processes is a legal requirement for health services. However, its high cost hinders its widespread use. Aiming at developing a cost-effective inoculum medium, soybean molasses and nutrient-supplemented vinasse were evaluated for their effectiveness in solid-state fermentation (SSF). In biomass production, the results demonstrated that all tested compositions favor growth by providing the nutritional demands of the microorganism. Optimum casein peptone and soybean molasses concentration (1.0%, 2.5%, or 4.0%) was determined by a 2(2–0) factorial experimental design. The results have showed a positive influence of peptone on biomass production. In order to define peptone final concentration (4.0% or 6.0%), a 22 factorial experimental design was used. An optimized medium containing 4.0% soybean molasses and 4.0% casein peptone was similar in performance to a synthetic control medium (tryptone soy broth) in dry-heat thermal-resistant spore production by SSF. An experiment performed under optimum SSF conditions resulted in 1.9 × 1010 CFU g−1 dry matter with D 160 °C = 5.2 ± 0.2 min.

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Acknowledgments

The present research was financially supported by the Secretaria de Estado da Ciência, Tecnologia e Ensino Superior—Fundo Paraná. The authors thank José Carlos Pereira Comink for the statistic revision and Neuza Araujo and Elza Retka for the laboratory support.

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Authors and Affiliations

  1. Secretaria de Estado da Saúde do Paraná, Centro de Produção e Pesquisa de Imunobiológicos, Av.São Roque, 716, 83. 302-200, Piraquara, PR, Brazil

    Sandra Regina B. R. Sella & Regina Elizabete F. Dlugokenski

  2. Divisão de Engenharia de Bioprocessos & Biotecnologia, Universidade Federal do Paraná, Caixa Postal 19031, 81.531-970, Curitiba, PR, Brazil

    Sandra Regina B. R. Sella, Regina Elizabete F. Dlugokenski, Belquis P. Guizelini, Luciana P. S. Vandenberghe, Adriane B. P. Medeiros & Carlos Ricardo Soccol

  3. Regional Research Laboratory, Biotechnology Division, CSIR, Trivandrum, 695 019, India

    Ashok Pandey

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  1. Sandra Regina B. R. Sella
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  2. Regina Elizabete F. Dlugokenski
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  3. Belquis P. Guizelini
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  4. Luciana P. S. Vandenberghe
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Correspondence to Sandra Regina B. R. Sella.

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Sella, S.R.B.R., Dlugokenski, R.E.F., Guizelini, B.P. et al. Selection and Optimization of Bacillus atrophaeus Inoculum Medium and its Effect on Spore Yield and Thermal Resistance. Appl Biochem Biotechnol 151, 380–392 (2008). https://doi.org/10.1007/s12010-008-8206-3

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  • DOI: https://doi.org/10.1007/s12010-008-8206-3

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