Batch CGTase Production with Free and Immobilized <i>Bacillus firmus</i>Strain 37 in Bovine Bone Charcoal

The present study aimed to study the batch production of CGTase (cyclomaltodextrin-glucanotransferase) with Bacillus firmus strain 37 free and immobilized in bovine bone charcoal in batch mode and in a fluidized bed batch reactor, respectively. The bovine bone charcoal is an innovative support material for the immobilization of microorganisms’ producers of enzymes and the use of this microbial support allows its reuse to a significant cost reduction of the process. The batch fermentation with free cells was investigated for 96 h and reached a CGTase activity equal to 0.77 U/mL. When the microorganism was immobilized on bovine bone charcoal (7 g) and cultivated in fluidized bed batch reactor with air supplementation (1 volume of air/volume of medium * minute), the same activity could be achieved in 24 h. The results of enzymatic activity achieved show the potential of CGTase production in a short time with Bacillus firmus strain 37 immobilized in bovine bone charcoal matrix and using air supplementation in the production medium.

Bovine Bone Charcoal as Support Material for Immobilization of <em>Bacillus firmus</em>Strain 37 and Production of Cyclomaltodextrin Glucanotransferase by Batch Fermentation in a Fluidized Bed

The process described in the present work uses air supplementation in a fluidized bed reactor containing Bacillus firmus strain 37 immobilized on active bovine bone charcoal, to produce by batch fermentation the enzyme CGTase (cyclomaltodextrin-glucanotransferase). Three different aeration rates were evaluated. The maximum CGTase activity was achieved after 120 hours of fermentation with aeration rate of 2 vvm and was equal to 2.48 U/mL. When 0.5 and 1 vvm were used the enzymatic activities achieved 1.1 and 0.57 U/mL, respectively. Bovine bone charcoal was characterized in terms of surface area, pore size and volume. To the best of our knowledge, the immobilization of microorganism cells in bovine bone charcoal for CGTase production has not been reported in the literature. Our results showed that fluidized bed reactor allows retaining high concentration of biomass, improving biomass-substrate contact and operation at low residence times, which resulted in improved enzyme production. Therefore, the process as proposed has great potential for industrial development.