E. Gomes

Optimization of xylanase production by Bacillus circulans D1 in submerged fermentation using response surface methodology

Abstract In this work, a 3 3 factorial design was performed with the aim of optimizing the culture conditions for xylanase production by an alkalophilic thermophilic strain of Bacillus circulans , using response surface methodology. The variables involved in this study were xylan concentration ( X 1 ), pH ( X 2 ) and cultivation time ( X 3 ). The optimal response region was approached without using paths of steepest ascent. Statistical analysis of results showed that, in the range studied, only pH did not have a significant effect on xylanase production. A second-order model was proposed to represent the enzymic activity as a function of xylan concentration ( X 1 ) and cultivation time ( X 3 ). The optimum xylan concentration and cultivation time were 5 g/l and 48 h, respectively. Under these conditions, the model predicted a xylanase activity of 19.1 U/ml.

Application of crude xylanase from Bacillus licheniformis 77-2 to the bleaching of eucalyptus Kraft pulp

Alkalophilic Bacillus licheniformis 77-2 produced an extracellular alkali-tolerant xylanase with negligible cellulase activity in medium containing corn straw. The effectiveness of crude xylanase on treatment of eucalyptus Kraft pulp was evaluated. A biobleaching experiment was carried out to compare the chlorine saving with pulp treated and untreated by the enzyme. Two-stage bleaching was employed, using a ClO2 chlorination and NaOH extraction (DE sequence). With the enzymatic treatment, in order to obtain the same value of Kappa number and brightness, respectively 28.5 and 30% less ClO2 was required in comparison to the enzymatically untreated samples.

Purification and characterization of two beta-glucosidases from the thermophilic fungus Thermoascus aurantiacus

Abstractβ-Glucosidase from the fungusThermoascus aurantiacus grown on semi-solid fermentation medium (using ground corncob as substrate) was partially purified in 5 steps—ultrafiltration, ethanol precipitation, gel filtration and 2 anion exchange chromatography runs, and characterized. After the first anion exchange chromatography, β-glucosidase activity was eluted in 3 peaks (Gl-1, Gl-2, Gl-3). Only the Gl-2 and Gl-3 fractions were adsorbed on the gel matrix. Gl-2 and Gl-3 exhibited optimum pH at 4.5 and 4.0, respectively. The temperature optimum of both glucosidases was at 75–80°C. The pH stability of Gl-2 (4.0–9.0) was higher than Gl-3 (5.5–8.5); both enzyme activities showed similar patterns of thermostability. Under conditions of denaturing gel chromatography the molar mass of Gl-2 and Gl-3 was 175 and 157 kDa, respectively. Using 4-nitrophenyl β-d-glucopyranoside as substrate,Km values of 1.17±0.35 and 1.38±0.86 mmol/L were determined for Gl-2 and Gl-3, respectively. Both enzymes were inhibited by Ag+ and stimulated by Ca2+.

Pectinolytic enzyme production by Bacillus species and their potential application on juice extraction

Five Bacillus strains isolated from decaying vegetable material were cultivated on wheat bran and endo-polygalacturonases, exo-polygalacturonase and pectin lyase activities in the crude enzymatic solution obtained were determined. Highest activity was observed for all enzymes when fermentation was carried out at 28 °C, the highest activity values were obtained after 120 h of cultivation for exo-PG and after 48 h for endo-PG and PL. The use of the enzymatic solution for treatment of fruits and vegetable mash afforded a high juice extraction and a pulp with good pressing characteristics.

Thermophilic fungi as new sources for production of cellulases and xylanases with potential use in sugarcane bagasse saccharification

To obtain new cellulases and xylanases from thermophilic fungi; evaluate their potential for sugarcane bagasse saccharification.

Pectinase production by a Brazilian thermophilic fungus Thermomucor indicae-seudaticae N31 in solid-state and submerged fermentation

Thermophilic organisms produce thermostable enzymes, which have a number of applications, justifying the interest in the isolation of new thermophilic strains and study of their enzymes. Thirty-four thermophilic and thermotolerant fungal strains were isolated from soil, organic compost, and an industrial waste pile based on their ability to grow at 45°C and in a liquid medium containing pectin as the only carbon source. Among these fungi, 50% were identified at the genus level as Thermomyces, Aspergillus, Monascus, Chaetomium, Neosartoria, Scopulariopsis, and Thermomucor. All isolated strains produced pectinase during solid-state fermentation (SSF). The highest polygalacturonase (PG) activity was obtained in the culture medium of thermophilic strain N31 identified as Thermomucor indicae-seudaticae. Under SSF conditions on media containing a mixture of wheat bran and orange bagasse (1 : 1) at 70% of initial moisture, this fungus produced the maximum of 120 U/ml of exo-PG, while in submerged fermentation (SmF) it produced 13.6 U/ml. The crude PG from SmF was more thermostable than that from SSF and exhibited higher stability in acidic pH.

Screening for pectinolytic activity of wood-rotting basidiomycetes and characterization of the enzymes

Seventy-five fungal strains from different groups of basidiomycetes, newly isolated from rotten wood, were screened for pectinolytic activity. Despite the fact that basidiomycetes are scarcely referred to as pectinase producers, the polygalacturonase (PG) activity was detected in 76 % of the strains; 16 % with activity higher than 40 nkat/g, 40 % between 13.3 and 40 nkat/g, and 44 % with activity lower than 13.3 nkat/g. The highest productions were obtained among the fungi from orderAphyllophorales, familyPolyporaceae. The characterization of the enzymes from the highest PG producers (Lentinus sp.,Gloeophyllum striatum, Pycnoporus sanguineus, Schizophyllum commune) showed optimum temperature for catalytic activity at 60–70°C and two peaks of pH optimum (3.5–4.5 and 8.5–9.5). The enzymes exhibited high pH stability (3.0–11.0) but after incubation at 40°C for 1 h their activity dropped by 18–73%.

Endoglucanase production with the newly isolated Myceliophtora sp. i-1d3b in a packed bed solid state fermentor

This work is aimed to produce endoglucanase through solid state fermentation in a packed bed bioreactor with the use of the fungus Myceliophtora sp. I-1D3busing a mixture of wheat bran (WB) and sugar cane bagasse (SCB) as culture medium. Preliminary tests were performed in polypropylene plastic bags, controlling the variables temperature (40, 45, and 50oC), initial moisture content (75, 80, and 85%, w.b.), and weight proportion SCB/WB (1:1, 7:3, and 9:1). The highest enzyme activities in plastic bags were obtained using the substrate proportion of 7:3, 50oC temperature, and 80% initial moisture content (878 U/grams of dry solid). High activities of filter-paper cellulase and xylanase were also obtained in plastic bags and some results are reported. For the packed bed experiments, the temperature (45 and 50oC) and the air flow rate (80, 100 and 120L/h) were the controlled variables. Activity of endoglucanase was similar to plastic bag tests. A longitudinal gradient of moisture content, was observed increasing from the bottom to the top of the reactor, even though the longitudinal enzyme activity profile was flat for almost the whole bed. Air flow rate did not affect enzyme activity, while experiments carried out at 50oC showed higher enzyme activities. The maximum temperature peak observed was at about 6oC above the process temperature.

Production of cyclodextrins by CGTase from Bacillus clausii using different starches as substrates.

Cyclodextrins (CDs) are cyclic oligasaccharides composed by d-glucose monomers joined by α-1,4-d glicosidic linkages. The main types of CDs are α-, β- and γ-CDs consisting of cycles of six, seven, and eight glucose monomers, respectively. Their ability to form inclusion complexes is the most important characteristic, allowing their wide industrial application. The physical property of the CD-complexed compound can be altered to improve stability, volatility, solubility, or bio-availability. The cyclomaltodextrin glucanotransferase (CGTase, EC 2.4.1.19) is an enzyme capable of converting starch into CD molecules. In this work, the CGTase produced by Bacillus clausii strain E16 was used to produce CD from maltodextrin and different starches (commercial soluble starch, corn, cassava, sweet potato, and waxy corn starches) as substrates. It was observed that the substrate sources influence the kind of CD obtained and that this CGTase displays a β-CGTase action, presenting a better conversion of soluble starch at 1.0%, of which 80% was converted in CDs. The ratio of total CD produced was 0:0.89:0.11 for α/β/γ. It was also observed that root and tuber starches were more accessible to CGTase action than seed starch under the studied conditions.

Production, Characterization, and Properties of β-Glucosidase and β-Xylosidase from a Strain of Aureobasidium sp.

Abstractβ-Glucosidase and β-xylosidase production by a yeastlike Aureobasidium sp. was carried out during solid-state and submerged fermentation using different carbon sources and crude enzymes were characterized. β-Glucosidase and β-xylosidase exhibited optimum activities at pH 2.0–2.5 and 3.0, respectively. These enzymes had the maximum activities at 65°C and were stable in a wide pH range and at high temperatures.