Kátia Flávia Fernandes

Immobilization of horseradish peroxidase onto polyaniline polymers

Some chemically synthesized conductive polymers, based on polyaniline (PANI), were used as supports for Horseradish Peroxidase (HRP) immobilization. The polymers were first activated with glutaraldehyde (PANIG) and then HRP was successfully immobilized. Among the tested polymers, those with intermediate oxidation state (PANIG1, PANIG2 and PANIG3) were more suitable for HRP immobilization than that predominately reduced (PANIG2) or oxidized (PANIG5). PANIG1 presented the best performance for HRP immobilization, and the analysis of the protein versus active enzyme retention showed that immobilized HRP maintained 100% native activity. The effects of enzyme concentration, reaction time and pH on the immobilization efficiency were investigated.

Changes in the Functional Properties and Antinutritional Factors of Extruded Hard-to-Cook Common Beans (Phaseolus vulgaris, L.)

The biochemical and functional properties of 2 hard-to-cook common bean cultivars (Phaseolus vulgaris, L.) were investigated after the extrusion process. Beans of BRS pontal and BRS grafite cultivars were milled and extruded at 150 degrees C, with a compression ratio screw of 3 : 1, 5-mm die, and screw speed of 150 rpm. Extrudate flours were evaluated for water solubility (WS), water absorption index (WAI), oil absorption capacity (OAC), foaming capacity (FC), emulsifying activity (EA), antinutritional factors, and in vitro protein and starch digestibility. Results indicated that the extrusion significantly decreased antinutrients such as phytic acid, lectin, alpha-amylase, and trypsin inhibitors, reduced the emulsifying capacity and eliminated the FC in both BRS pontal and BRS grafite cultivars. In addition, the WS, WAI, and in vitro protein and starch digestibility were improved by the extrusion process. These results indicate that it is possible to produce new extruded products with good functional and biochemical properties from these common bean cultivars.

Biochemical characterization of selected plant species from Brazilian Savannas

The aim of this work was to analyze and quantify the presence of antinutritional compounds such as lectins and trypsin-like inhibitors, polyphenols and tannins, and enzymatic activity of peroxidases and proteases in the seeds of Annona crassiflora Mart. (araticum), Hymenaea courbaril L. var. courbaril (jatoba), Plathymenia reticulata Benth. (vinhatico), Zanthoxylum rhoifolium Lam. (maminha de porca), Apeiba tibourbou Aubl. (pau jangada), Salacia crassiflora Mart G. Don. (bacupari), and Sclerolobium paniculatum Vog. (carvoeiro). The results suggested that these plants could be used as new source of food.

Lipase entrapment in PVA/Chitosan biodegradable film for reactor coatings

This study reports the development and characterization of novel biodegradable film, based on chitosan and polyvinyl alcohol containing lipase entrapped. The films showed a thickness of 70.4 and 79 μm to PVA/Chitosan and PVA/Chitosan/Lipase, respectively. The entrapment of lipase in PVA/Chitosan film resulted in increasing of 69.4% tensile strength (TS), and 52.4% of elongation. SEM images showed the formation of a continuous film, without pores or cracks. The lipase entrapment efficiency was estimated in 92% and the films were repeatedly used for 25 hydrolytic cycles, maintaining 62% of initial activity. The PVA/Chitosan/Lipase film was used for olive oil hydrolysis of high performance. These results indicate that PVA/Chitosan/Lipase is a promising material for biotechnology applications such as triacylglycerol hydrolysis and biodiesel production.

Biodegradable and bioactive CGP/PVA film for fungal growth inhibition.

In this study, chitinolytic enzymes produced by Trichoderma asperellum were immobilized on a biodegradable film manufactured with a blend of cashew gum polysaccharide (CGP) and polyvinyl alcohol (PVA), and tested as a fungal growth inhibitor. The film was produced by casting a blend of CGP and PVA solution on glass molds. The CGP/PVA film showed 68% water solubility, tensile strength of 23.7 MPa, 187.2% elongation and 52% of mass loss after 90 days in soil. The presence of T-CWD enzymes immobilized by adsorption or covalent attachment resulted in effective inhibition of fungal growth. Sclerotinia sclerotiorum was the most sensitive organism, followed by Aspergillus niger and Penicillium sp. SEM micrograph showed that the presence of immobilized T-CWD enzymes on CGP/PVA film produced morphological modifications on vegetative and germinative structures of the microorganisms, particularly hyphae disruption and changes of spores shape.

Extraction and chemical characterization of starch from S. lycocarpum fruits

In this study the pulp from Solanum lycocarpum fruits was used as raw material for extraction of starch, resulting in a yield of 51%. The starch granules were heterogeneous in size, presenting a conical appearance, very similar to a high-amylose cassava starch. The elemental analysis (CHNS) revealed 64.33% carbon, 7.16% hydrogen and 0.80% nitrogen. FT-IR spectroscopy showed characteristic peaks of polysaccharides and NMR analysis confirmed the presence of the α-anomer of d-glucose. The S. lycocarpum starch was characterized by high value of intrinsic viscosity (3515 mPa s) and estimated molecular weight around 645.69 kDa. Furthermore, this starch was classified as a B-type and high amylose content starch, presenting 34.66% of amylose and 38% crystallinity. Endothermic transition temperatures (To=61.25 °C, Tp=64.5 °C, Tc=67.5 °C), gelatinization temperature (ΔT=6.3 °C) ranges and enthalpy changes (ΔH=13.21 J g(-1)) were accessed by DCS analysis. These results make the S. lycocarpum fruit a very promising source of starch for biotechnological applications.

Potential fungal inhibition by immobilized hydrolytic enzymes from Trichoderma asperellum.

The use of cell wall degrading enzymes from Trichoderma asperellum immobilized on biodegradable support is an alternative for food packaging. In this study, hydrolytic enzymes produced by T. asperellum were tested as a fungal growth inhibitor, in free form or immobilized on a biodegradable film composed of cassava starch and poly(butylene adipate-co-terephtalate) (PBAT). The inhibitory activity was tested against Aspergillus niger , Penicillium sp., and Sclerotinia sclerotiorum , microorganisms that frequently degrade food packaging. The use of chitin as carbon source in liquid medium induced T. asperellun to produce N-acetylglucosaminidase, β-1,3-glucanase, chitinase, and protease. The presence of T. asperellun cell wall degradating enzymes (T-CWD) immobilized by adsorption or covalent attachment resulted in effective inhibition of fungal growth. The enzymatic activity of T-CWD was stronger on S. sclerotiorum than on the Aspergillus or Penicillum isolates tested. These results suggest that T-CWD can be used in a free or immobilized form to suppress fungi that degrade food packaging.

Immobilization of Peroxidase onto Magnetite Modified Polyaniline

The present study describes the immobilization of horseradish peroxidase (HRP) on magnetite-modified polyaniline (PANImG) activated with glutaraldehyde. After the optimization of the methodology, the immobilization of HRP on PANImG produced the same yield (25%) obtained for PANIG with an efficiency of 100% (active protein). The optimum pH for immobilization was displaced by the effect of the partition of protons produced in the microenvironment by the magnetite. The tests of repeated use have shown that PANImG-HRP can be used for 13 cycles with maintenance of 50% of the initial activity.

A stimuli-responsive and bioactive film based on blended polyvinyl alcohol and cashew gum polysaccharide

In this study, a stimuli-responsive, biodegradable and bioactive film was produced by blending cashew gum polysaccharide (CGP) and polyvinyl alcohol (PVA). The film presented malleability and mechanical properties enabling an easy handling. Wetting the film changed the optical property from opacity to levels of transparency higher than 70% and resulted in up to 2-fold increase in its superficial area. Different swelling indexes were obtained varying the pH of solvent, which allows classifying the CGP/PVA film as pH sensitive stimuli-responsive material. The bioactivity was achieved through covalent immobilization of papain, which remained active after storage of CGP/PVA-papain film for 24h in the presence of buffer or in a dry form. These results evidenced that CGP/PVA-papain film is a very promising material for biomedical applications.

A bioactive film based on cashew gum polysaccharide for wound dressing applications.

This work presents the development of a new bioactive material for wound therapeutics which may play a dual role of modulate metallo proteinases activity while prevents infection blocking out pathogenic microorganisms and foreign materials. A CGP/PVA film was activated by covalent immobilization of trypsin. Results from biocompatibility test revealed that PDL fibroblasts grown on the surface of CGP/PVA and the high amount of viable cells proved absence of cytotoxicity. Trypsin immobilized onto CGP/PVA film remained 100% active after 28 days stored dried at room temperature. In addition, CGP/PVA-trypsin film could be used for 9 cycles of storage/use without loss of activity. After immobilization, trypsin retained its collagenolytic activity, indicating this material as a promising material for wound dressing applications.