Production of asporogenous mutants of Bacillus sphaericus 2362 in continuous culture

• Published in: Letters in applied microbiology Vol. 33; no. 1; pp. 40 - 44
• Main Authors: Idachaba, M.A., Rogers, P.L.
• Format: Journal Article
• Language: English
• Published: Oxford UK Blackwell Publishing Ltd 01.07.2001; Blackwell Science
• Subjects: Animals; Bacillus - genetics; Bacillus - growth & development; Bacillus - physiology; Bacillus sphaericus; Bacterial Toxins - biosynthesis; Bacterial Toxins - toxicity; Biological and medical sciences; Biological control; Biotechnology; Colony Count, Microbial; Control; Culex - microbiology; Culex - physiology; Culture Media; Fundamental and applied biological sciences. Psychology; Hot Temperature; Methods. Procedures. Technologies; Microbial engineering. Fermentation and microbial culture technology; Mutation; Phytopathology. Animal pests. Plant and forest protection; Protozoa. Invertebrates; Spores, Bacterial; Entomopathogen; Biological control; Bacillaceae; Bacillus sphaericus; Continuous process; Toxin; Bacillales; Sporulation; Production; Microorganism culture; Bacteria; Mutation; Biopesticide; Microbial insecticide
• ISSN: 0266-8254; 1472-765X; 1365-2673
• DOI: 10.1046/j.1472-765X.2001.00955.x

Aims: To report the production of asporogenous mutants (Spo−) of Bacillus sphaericus 2362 in continuous culture. Methods and Results: Microbial culture samples were taken at 0.05 h−1 dilution rate and plated out on nutrient agar plates. Translucent colonies were obtained with vegetative morphology under phase contrast microscope. Heat resistance evaluations at different temperature settings showed that the Spo− mutants had lower heat resistance than the Spo+ wild type. Western blots analyses carried out on both wild type and the mutants indicated the presence of binary protein toxins of 42 and 51 kDa in both. Bioassays carried out on the wild type and the Spo– mutants against mosquitoes showed the mutants to be 100‐fold less toxic in comparison to the wild type. Conclusions: Existence and production of asporogenous mutants of Bacillus sphaericus 2362 in continuous culture at low dilution rates is demonstrated by this study. The organism's ability to produce toxins appears to be significantly reduced by the mutational process. Significance and Impact of the Study: The production of asporogenous mutants had not been reported previously among strains of Bacillus sphaericus. The present report on the toxigenic capability of asporogenous mutants also raises the possibility of using continuous culture to significantly improve the productivity of toxin production in future.