The immobilization of horseradish peroxidase (HRP) on raw and alkaline pre-treated sugarcane bagasse by physical adsorption (ADS) and covalent bond (LC) methods was studied. The saturation of the support with 2 mg of HRP/g of support by LC immobilization reached 35% of immobilization efficiency and 39 units of the immobilized enzyme (U). Regarding the HRP immobilization on sugarcane bagasse without pretreatment and using the same HRP loading, it was observed a reduction in the efficiency of immobilization and in the number of immobilized units for both methods, ADS (13.98% and 15.46 U) and LC (15.79% and 17.46 U). The sugarcane bagasse with alkaline pretreatment experiment, on the other hand, exhibited higher potential for HRP immobilization by LC. The supports and biocatalysts were characterized by Fourier transform infrared spectroscopy (FTIR), showing greater availability of hydroxyl groups in the pretreated support and the typical amide I and amide II bands that corroborate the effectiveness of the enzyme immobilization on sugarcane bagasse. In the same way, the thermogravimetric analysis (TGA) confirmed a higher weight loss in the region I for the derivative immobilized by LC, suggesting the presence of water favored enzymatic activity.

Mesoporous silica supports (aerogels) were used to immobilize Burkholderia cepacia lipase (BC) by encapsulation (EN or ENIL), physical adsorption (ADS or ADSIL) and covalent binding (CB or CBIL) into or onto the aerogel modified with protic ionic liquid (PIL). Yield immobilization (Ya) and operational stability were determined by the hydrolytic reaction of olive oil. Ya (37% to 83% by physical adsorption) and operational stability (2 to 23 batches by encapsulation) increased when the support was modified with PIL. For immobilized derivates observed by the BET method, in this case ADS and CB for ADSIL and CBIL, increased pores size was observed, possibly due to the higher amount of BC immobilized conferring Ya and operational stability. This effect was probably attributed to the entry of the enzyme into the pores of the silica aerogel structure. SEM images showed a change in the structure and properties of immobilized lipase derived with PIL. A characteristic FTIR band was obtained for the silanol groups and amides I, IV and V, demonstrating the efficiency of immobilization of BC. The most efficient biocatalysts were ADSIL with regard to yield immobilization and ENIL for operational stability.

This work investigated the effect of microwave irradiation (MW) on the ethanolysis rate of soybean and sunflower oils catalyzed by supported Novozyme 435 (Candida antarctica). The effects of tert-butanol, water addition and oil:ethanol molar ratio on transesterification were evaluated under conventional heating (CH), and under optimum reaction conditions (with no added water in the system, 10% tert-butanol and 3:1 ethanol-to-oil molar ratio). The reactions were monitored up to 24 h to determine the conditions of initial reaction velocity. The investigated variables under MW (50 W) were: reaction time (5.0-180 min) and mode of reactor operation (fixed power, dynamic and cycles) in the absence and presence of tert-butanol (10% (w/w). The measured response was the reaction conversion in ethyl esters, which was linked to the enzyme catalytic activity. The results indicated that the use of microwave improved the activity at fixed power mode. A positive effect of the association of tert-butanol and MW irradiation on the catalytic activity was observed. The reaction rate improved in the order of approximately 1.5 fold compared to that under CH with soybean oil. Using soybean oil, the enzymatic transesterification under MW for conversion to FAEE (fatty acid ethyl esters) reached >99% in 3h, while with the use of CH the conversions were about 57% under similar conditions.

Lipase from Burkholderia cepacia was immobilized in a silica matrix and dried in high pressure carbon dioxide media (aerogel). The protic ionic liquid (PIL) was used in the immobilization process by encapsulation. The objective of this work was to evaluate the influence of the drying technique using supercritical carbon dioxide in biocatalysts obtained through the sol-gel technique by evaluating temperature and pressure and, after selecting the best drying conditions, to investigate the application of the technique for the biocatalyst using ionic liquid as an additive in the immobilization process. The results for immobilized biocatalysts showed that the best conditions of pressure and temperature were 100 bar and 25 ºC, respectively, giving a total activity recovery yield of 37.27% without PIL (EN) and 44.23% with PIL (ENLI). The operational stability of the biocatalysts showed a half-life of 11.4 h for ENLI and 6 h for EN. Therefore, solvent extraction using supercritical CO2, besides shortening drying time, offers little resistance to the immobilization of lipases, since their macropores provide ample room for their molecules. The use of the ionic liquid as an additive in the process studied for the immobilization of enzymes produced attractive yields for immobilization and therefore has potential for industrial applications in the hydrolysis of vegetable oils.

This work presents biochemical characterization of a lipase from a new strain of Bacillus sp. ITP-001, immobilized using a sol gel process (IB). The results from the biochemical characterization of IB showed increased activity for hydrolysis, with 526.63 U g-1 at pH 5.0 and 80 ºC, and thermal stability at 37 ºC. Enzymatic activity was stimulated by ions such as EDTA, Fe+3, Mn+2, Zn+2, and Ca+2, and in various organic solvents. Kinetic parameters obtained for the IB were Km = 14.62 mM, and Vmax = 0.102 mM min-1 g-1. The results of biochemical characterization revealed the improved catalytic properties of IB.

Lipases are characterised mainly by catalytic versatility and application in different industrial segments. The aim of this study was to biochemically characterise a lipase from a new strain of Bacillus sp. ITP-001. The isoelectric point and molecular mass were 3.12 and 54 kDa, respectively. The optima lipase activity was 276 U g-1 at pH 7.0 and a temperature of 80 ºC, showing greater stability at pH 5.0 and 37 ºC. Enzymatic activity was stimulated by various ions and pyridine, and inhibited by Cu+ and ethanol. The values of Km and v max were 105.26 mmol and 0.116 mmol min-1 g-1, respectively determined by the Eadie-Scatchard method.