Optimization of the cultivation conditions of Bacillus licheniformis BCLLNF-01 for cellulase production.

The objective of this study was to optimize the production of CMCase by Bacillus licheniformis BCLLNF-01, a strain associated with the mucus of the zoanthid Palythoa caribaeorum (Cnidaria, Anthozoa). Production of total cellulase and CMCase was investigated in the supernatant, intracellular content and wall content. Cultivation was carried out in BLM medium supplemented with 1.5 % (w/v) CMC, 5.5 % (v/v) inoculum, 40 °C, pH 6.5, 500 rpm for 72 h, and the highest activity was recorded in the supernatant. A Rotational Central Composite Design (RCCD) 2³ was used to investigate the influence of the carbon source concentration (CMC-0.5 to 1.5 % w/v), inoculum concentration (1-10 % v/v) and temperature (35-45 °C) on CMCase production. The maximum enzyme activity was achieved for a CMC concentration of 1.5 % w/v at 40 °C, attaining 0.493 IU/mL after 96 h of cultivation.

Production of concentrates of bacterial bio-insecticide Bacillus thuringiensis var. israelensis by flocculation/sedimentation.

Flocculation/sedimentation of Bacillus thuringiensis var. israelensis (Bti) using flocculating agents has been studied. Batch cell production was performed in an agitated tank, and the flocculation assays were carried out in jar tests. Flocculent suspensions were characterized based on diameter of flocs and density. The best results were obtained with CaCl(2).2H(2)O, FeCl(3).6H(2)O, Al(2)(SO(4))(3) and tannin, with optimal flocculation concentrations of 2500, 2500, 3500 and 1000 mg l(-1), respectively. Thickening of the flocculent suspensions was investigated, leading to determination of the capacity curves of the settler. Bioassays against Aedes aegypti larvae demonstrated excellent results in insect control.

Concentration of Cupriavidus necator cells by flocculation and sedimentation.

Separation and cells concentration constitute important stages in most biotechnological processes. Particularly, use of flocculation/sedimentation can improve significantly the extraction of biopolymers accumulated by microorganisms and the biodegradation of xenobiotic compounds by cell sludge. In this work the use of tannin and aluminum sulphate (Al2(SO4)3) as flocculating agents for concentration of cells of Cupriavidus necator DSM 545 is evaluated. Cells were grown in broth nutrient medium in Erlenmeyer flasks, submitted to orbital agitation of 160 rpm at 30 °C for 21 h. The optimal concentrations of flocculating agents, as determined with a standard jar test method, were equal to 2,800 mg/L for tannin and 800 mg/L for Al2(SO4)3, allowing for recovery of 95% of the cells in both cases. Obtained flocs presented density and average diameter of 1.03 g/mL ± 0.01 g/mL and 158 µm ± 19 µm for tannin and of 1.05 g/mL ± 0.01 g/mL and 146 µm ± 14 µm for Al2(SO4)3, respectively. Batch settling tests were performed in order to determine the operational capacity of continuous settlers to be used for separation of the investigated flocculent suspensions. Finally, cultivation of cells using flocs as inoculum indicated that the cells remained viable after flocculation with usage of the optimum flocculating agent concentrations.