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KMID : 1094720130180010075
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Biotechnology and Bioprocess Engineering
2013 Volume.18 No. 1 p.75 ~ p.87
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Soybean molasses-based bioindicator system for monitoring sterilization process: Designing and performance evaluation
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Sella Sandra Regina B. R.
Masetti Carla
Figueiredo Luis Felipe M.
Vandenberghe Luciana P. S.
Minozzo Joao Carlos
Soccol Carlos Ricardo
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Abstract
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A novel cost-effective Bacillus atrophaeus Sterilization Bioindicator System (BIS) with a high quality and performance was developed from a soybean byproduct and compared with the commercial BIS. It was composed of recovery medium and dry-fermented spores with sand as the support. The BIS was developed and optimized using a sequential experimental design strategy. The recovery medium contained soluble starch (1.0 g/L), soybean molasses (30.0 g/L), tryptone (40.0 g/L), and bromothymol blue (0.02 g/L) at pH 8.5. The solid-state fermentation conditions of the bioreactor and environmental humidity had no significant effects on the spore yield and dry-heat resistance. The only substrate mineral that showed a positive effect was Mn2+, allowing Mg2+, K+, and Ca2+ to be eliminated from the formulation. Validation of optimized medium indicated D 160°C = 6.8±1.0 min (3.6 min more than the minimum) and spore yield = 2.3 ± 0.5 × 109 CFU/g dry sand (10,000 × initial values). BIS performance resulted in D 160°C = 6.6 ± 0.1 min. Sporulation and germination kinetics allowed the sporulation process to be reduced to three days, and the growth of heat-damaged spores was sufficient to achieve visual identification of a non-sterile BIS within 21 h. Process economics was a minimum of 23.9%, and process cycle time was reduced from 29 to 15 days. The new BIS parameters demonstrated compliance to all regulatory requirements. No studies have yet described a BIS production from soybean molasses.
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KEYWORD
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Bacillus atrophaeus, solid-state fermentation, bioindicator system, soybean molasses, recovery medium, sterilization
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