Bacillus subtilis Endospores at High Purity and Recovery Yields: Optimization of Growth Conditions and Purification Method

• Published in: Current microbiology Vol. 66; no. 3; pp. 279 - 285
• Main Authors: Tavares, Milene B, Souza, Renata D, Luiz, Wilson B, Cavalcante, Rafael C. M, Casaroli, Caroline, Martins, Eduardo G, Ferreira, Rita C. C, Ferreira, Luís C. S
• Format: Journal Article
• Language: English
• Published: New York Springer-Verlag 01.03.2013; Springer Nature B.V
• Subjects: antigens; Bacillus subtilis; Bacillus subtilis - cytology; Bacillus subtilis - growth & development; Bacillus subtilis - physiology; Bacteria; Biomedical and Life Sciences; Biotechnology; Culture Media; Data processing; endospores; Growth conditions; Growth media; Life Sciences; lysozyme; Microbiology; probiotics; purification methods; qualitative analysis; Spores, Bacterial - cytology; Spores, Bacterial - isolation & purification; sporulation; Time Factors; Subtilis Spore; Spore Yield; Bacillus Spore; Spore Concentration; Recovery Yield
• ISSN: 0343-8651; 1432-0991
• DOI: 10.1007/s00284-012-0269-2

Bacillus subtilis endospores have applications in different fields including their use as probiotics and antigen delivery vectors. Such specialized applications frequently require highly purified spore preparations. Nonetheless, quantitative data regarding both yields and purity of B. subtilis endospores after application of different growth conditions and purification methods are scarce or poorly reported. In the present study, we conducted several quantitative and qualitative analyses of growth conditions and purification procedures aiming generation of purified B. subtilis spores. Based on two growth media and different incubations conditions, sporulation frequencies up to 74.2 % and spore concentrations up to 7 × 10⁹ spores/ml were achieved. Application of a simplified spore isolation method, in which samples were incubated with lysozyme and a detergent, resulted in preparations with highly purified spores at the highest yields. The present study represents, therefore, an important contribution for those working with B. subtilis endospores for different biotechnological purposes.