No abstract

Bacteria isolated from thermophilic environment that can produce cellulase as well as utilise agro-waste biomass have a high potential for developing thermostable cellulase required in the biofuel industry. The cost for cellulase represents a significant challenge in converting lignocellulose to fermentable sugars for biofuel production. Among three potential bacteria examined, Bacillus licheniformis 2D55 (accession no. KT799651) was found to produce the highest cellulolytic activity (CMCase 0.33 U/mL and FPase 0.09 U/mL) at 18–24 h fermentation when grown on microcrystalline cellulose (MCC) as a carbon source in shake flask at 50 °C. Cellulase production process was further conducted on the untreated and NaOH pretreated rice straw (RS), rice husk (RH), sugarcane bagasse (BAG) and empty fruit bunch (EFB). Untreated BAG produced the highest FPase (0.160 U/mL), while the highest CMCase (0.150 U/mL) was supported on the pretreated RH. The mixture of untreated BAG and pretreated RH as agro-waste cocktail has remarkably improved CMCase (3.7- and 1.4-fold) and FPase (2.5- and 11.5-fold) compared to the untreated BAG and pretreated RH, respectively. The mechanism of cellulase production explored through SEM analysis and the location of cellulase enzymes of the isolate was also presented. Agro-waste cocktail supplementation provides an alternative method for an efficient production of cellulase.

Effects of nutritional and physicochemical factors were investigated for cellulase production by the newly isolated thermophilic strain Bacillus licheniformis 2D55 (Accession No. KT799651). The optimum cellulase production in shake flask fermentation was attained at 60 °C, pH 3.5, 180 rpm, and in a medium containing untreated sugarcane bagasse and pretreated rice husk at 7% (w/v), urea, 1 g/L, peptone, 11.0 g/L, Mg(SO4)2, 0.40 g/L, CaCl2, 0.03 g/L, Tween 80, 0.2% (w/v), and 3% inoculum. The highest carboxymethyl cellulase (CMCase), filtre paperase (FPase), and β-glucosidase produced under the optimized conditions were 29.4 U/mL, 12.9 U/mL, and 0.06 U/mL, respectively, after 18 h of fermentation. Optimization of the parameters increased the CMCase, FPase, and β-glucosidase activities by 77.4-fold, 44.5-fold, and 10-fold, respectively. The crude enzyme was highly active and stable over broad temperature (50 to 80 °C) and pH (3.5 to 10.0) ranges with optimum temperature at 65 °C and 80 ºC for CMCase and FPase, respectively. The optimum pH for CMCase and FPase was 7.5 and 6.0, respectively. Saccharification of sugar cane bagasse and rice husk by crude cellulase resulted in perspective yields of 0.348 and 0.301 g g -1 dry substrate of reducing sugars. These results suggest prospects of thermostable cellulase from B. licheniformis 2D55 in application for bio-sugar production and other industrial bioprocess applications involving high temperatures.

Submerged substrate fermentation of Jatropha seed cake, a by-product of oil extraction from Jatropha curcas seed was carried out using Aspergillus terreus for the production of itaconic acid. The Jatropha seed cake was initially converted into fermentable sugars by dilute acid hydrolysis using 50% sulphuric acid. The rate of hydrolysis was 1.04 gL -1 . The fermentation process was carried out at room temperature, agitation of 400 rpm and three physico-chemical parameters (pH, inoculum size and substrate concentration) were varied. Itaconic acid and glucose assays were carried out by spectrophotometry and Dinitrosalicylic acid methods respectively daily. Maximum yield of itaconic acid was 48.70 gL -1 at 5 ml of inoculum size, 50% substrate concentration and pH 1.5. The residual glucose concentration increased for the first two days of fermentation after which it began to decrease as the itaconic acid concentration increased. The least concentration of itaconic acid observed was 6.00 gL -1 , obtained after 24 hours of fermentation with 4 ml inoculum size, 50% substrate concentration and at pH 1.5. The findings of this work indicate that Jatropha curcas seed cake is a suitable substrate for itaconic acid production.