A review on the immobilization of pepsin: A Lys-poor enzyme that is unstable at alkaline pH values.

Pepsin is a protease used in many different applications, and in many instances, it is utilized in an immobilized form to prevent contamination of the reaction product. This enzyme has two peculiarities that make its immobilization complex. The first one is related to the poor presence of primary amino groups on its surface (just one Lys and the terminal amino group). The second one is its poor stability at alkaline pH values. Both features make the immobilization of this enzyme to be considered a complicated goal, as most of the immobilization protocols utilize primary amino groups for immobilization. This review presents some of the attempts to get immobilized pepsin biocatalyst and their applications. The high density of anionic groups (Asp and Glu) make the anion exchange of the enzyme simpler, but this makes many of the strategies utilized to immobilize the enzyme (e.g., amino-glutaraldehyde supports) more related to a mixed ion exchange/hydrophobic adsorption than to real covalent immobilization. Finally, we propose some possibilities that can permit not only the covalent immobilization of this enzyme, but also their stabilization via multipoint covalent attachment.

Effect of heat treatment on oxalate and hydrocyanic acid levels of malanga corms of two cultivars (Xanthosoma sagittifolium and Colocasia esculenta) in a murine model.

Xanthosoma sagittifolium and Colocasia esculenta contain high levels of nutrients; but have naturally toxic compounds, oxalates and hydrocyanic acid (HCN). The objective of this work was to evaluate the effect of heat treatment on the concentration of antinutrients in malanga corms and its effect on mice. Malanga samples were heated to a boil for 0 to 120 min; oxalates and HCN were determined by spectrophotometry, at 710 and 510 nm, respectively. Pellets were prepared from raw malanga flour (15 and 50%), cooked malanga (15 and 50%) and wheat flour (control) and fed for nine weeks to five groups of six mice each. Cooking of X. sagittifolium corms for 80 min reduced oxalates present by 75% (143 to 35.6 mg/100 g sample), while oxalates in C. esculenta were reduced by 83% (345 to 57.8 mg/100 g sample). HCN levels became negligible after 20 min of cooking. During the nine weeks of feeding the different mice groups showed no significant difference (p > 0.05) between initial and final weight, with respect of the control; mice did not lose their appetite. The results indicate that the consumption of cooked malanga does not pose an evident risk to health, assessed by the reduced level of antinutrients, being an excellent alternative for feeding people in communities with prevalence of food insecurity.

Physicochemical Characterization of Resistant Starch Type-III (RS3) Obtained by Autoclaving Malanga (Xanthosoma sagittifolium) Flour and Corn Starch.

The feasibility of obtaining resistant starch type III (RS3) from malanga flour (Xanthosoma sagittifolium), as an unconventional source of starch, was evaluated using the hydrothermal treatment of autoclaving. The physicochemical characterization of RS3 made from malanga flour was carried out through the evaluation of the chemical composition, color attributes, and thermal properties. In addition, the contents of the total starch, available starch, resistant starch, and retrograded resistant starch were determined by in vitro enzymatic tests. A commercial corn starch sample was used to produce RS3 and utilized to compare all of the analyses. The results showed that native malanga flour behaved differently in most of the evaluations performed, compared to the commercial corn starch. These results could be explained by the presence of minor components that could interfere with the physicochemical and functional properties of the flour; however, the RS3 samples obtained from malanga flour and corn starch were similar in their thermal and morphological features, which may be related to their similarities in the content and molecular weight of amylose, in both of the samples. Furthermore, the yields for obtaining the autoclaved powders from corn starch and malanga flour were similar (≈89%), which showed that the malanga flour is an attractive raw material for obtaining RS3 with adequate yields, to be considered in the subsequent research.

Bioactive peptides from fisheries residues: A review of use of papain in proteolysis reactions.

Papain is a cysteine endopeptidase of vegetal origin (papaya (Carica papaya L.) with diverse applications in food technology. In this review we have focused our attention on its application in the production of bio-peptides by hydrolysis of proteins from fish residues. This way, a residual material, that can become a contaminant if dumped without control, is converted into highly interesting products. The main bioactivity of the produced peptides is their antioxidant activity, followed by their nutritional and functional activities, but peptides with many other bioactivities have been produced. Thera are also examples of production of hydrolysates with several bioactivities. The enzyme may be used alone, or in combination with other enzymes to increase the degree of hydrolysis.

Chitosan Films Obtained from Brachystola magna (Girard) and Its Evaluation on Quality Attributes in Sausages during Storage.

High molecular weight chitosan (≈322 kDa) was obtained from chitin isolated from Brachystola magna (Girard) to produced biodegradable films. Their physicochemical, mechanical and water vapor permeability (WVP) properties were compared against commercial chitosan films with different molecular weights. Brachystola magna chitosan films (CFBM) exhibited similar physicochemical and mechanical characteristics to those of commercial chitosans. The CFBM films presented lower WVP values (10.01 × 10-11 g/m s Pa) than commercial chitosans films (from 16.06 × 10-11 to 64.30 × 10-11 g/m s Pa). Frankfurt-type sausages were covered with chitosan films and stored in refrigerated conditions (4 °C). Their quality attributes (color, weight loss, pH, moisture, texture and lipid oxidation) were evaluated at 0, 5, 10, 15 and 20 days. Sausages covered with CFMB films presented the lowest weight loss (from 1.24% to 2.38%). A higher increase in hardness (from 22.32 N to 30.63 N) was observed in sausages covered with CFMB films. Compared with other films and the control (uncovered sausages), CFMB films delay pH reduction. Moreover, this film presents the lower lipid oxidation level (0.10 malonaldehyde mg/sample kg). Thus, chitosan of B. magna could be a good alternative as packaging material for meat products with high-fat content.

Lipoxygenase and Its Relationship with Ethylene During Ripening of Genetically Modified Tomato (Solanum lycopersicum).

RESEARCH BACKGROUND: TomloxB is the main isoform of lipoxygenase associated with ripening and senescence of fruits. On the other hand, ethylene, a gaseous hormone, is essential for the regulation of ripening in climacteric fruits like tomatoes. However, the relationship between TomloxB and ethylene production has not been thoroughly studied. Therefore, we aim to assess the effect of exogenous ethylene in transgenic tomatoes that contain a silenced TomloxB gene, and subsequently evaluate lipoxygenase activity, 1-aminocyclopropane-1-carboxylic acid oxidase and ethylene production; as well as to quantify the expression of the genes encoding 1-aminocyclopropane-1-carboxylic acid oxidase and TomloxB. EXPERIMENTAL APPROACH: To investigate the effect of lipoxygenase and 1-aminocyclopropane-1-carboxylic acid oxidase activity, fruits harvested at the stages of break, turning and pink were used. Tomatoes at break stage collected from transgenic and wild type plants were used to determine ethylene production and gene expression. Genetically modified and wild type tomato fruits were exposed to 100 µL/L exogenous ethylene. Lipoxygenase activity was measured spectrophotometrically. Activity of 1-aminocyclopropane-1-carboxylic acid oxidase and ethylene production were determined by gas chromatography. Oligonucleotides for differentially expressed genes: 1-aminocyclopropane-1-carboxylic acid oxidase and TomloxB were used to determine gene expression by real-time PCR. RESULTS AND CONCLUSIONS: The data showed that silencing of TomloxB caused a reduction in lipoxygenase activity and ethylene production in tomato fruits, and also reduced 1-aminocyclopropane-1-carboxylic acid oxidase activity. Hence, the addition of exogenous ethylene increased lipoxygenase activity in all treatments and 1-aminocyclopropane-1-carboxylic acid oxidase activity only in transgenic lines at break stage, consequently there was a positive regulation between TomloxB and ethylene, as increasing the amount of ethylene increased the activity of lipoxygenase. The results suggest that lipoxygenase may be a regulator of 1-aminocyclopropane-1-carboxylic acid oxidase and production of ethylene at break stage. NOVELTY AND SCIENTIFIC CONTRIBUTION: These results lead to a better understanding of the metabolic contribution of TomloxB in fruit ripening and how it is linked to the senescence-related process, which can lead to a longer shelf life of fruits. Understanding this relationship between lipoxygenase and ethylene can be useful for better post-harvest handling of tomatoes.

Increased Postharvest Life of TomLox B Silenced Mutants of Tomato (Solanum lycopersicum) Var. TA234.

A healthy lifestyle includes fruits and vegetables consumption. Tomato is one of the most consumed vegetables, although it is susceptible to physical damage through postharvest handling, thus leading to important losses. Softening is an important variable during tomato ripening; excessive softening is undesirable and leads to postharvest losses. TomloxB plays an important role in ripening, mainly in the loss of cellular integrity caused by fatty acids released from the lipid matrix of membranes that initiate oxidative deterioration, which is in turn carried into senescence. In order to increase postharvest life, we produced transgenic tomato plants via Rhizobium radiobacter with tomato lipoxygenase B (TomloxB) antisense constructs under control of the cauliflower mosaic virus (CaMV) 35S promoter. Lipoxygenase activity and firmness were measured in tomato fruit and the fatty acids profile was determined. Transgenic fruits were maintained for 40 days at room temperature in optimal conditions, whereas wild type fruits remained in similar conditions for only six days. Firmness in pink and red stages was significantly lower in wild type fruits than in two transgenic lines. Linolenic acid was the most important fatty acid consumed by lipoxygenase in both turning and pink stages of ripening. Lipoxygenase activity was smaller in transformed fruits in comparison with the wild type. These results suggest that silencing the TomloxB gene promoted significant changes in the physiology of transformed tomatoes, being the increase in postharvest life the most important.

Evaluación nutricional y sensorial de un alimento a base de atún y soya enriquecido con vitaminas y minerales / Nutritional and sensorial evaluation of a food with tuna and soya enriched with vitamins and minerals

The objective of this research was to determine the nutritional and sensorial impact of a food with tuna and soya enriched with vitamins and minerals in pre-school children of three communities of Chiapas, Mexico. It is a study of nutritional intervention with a cohorte of six months. The nutritional state of children was determined previuos to the nurishing complement, to the half and at the end of this. The sample was conformed with the children who presented undernourishment problems; they were fed during six months. The level of acceptability of the food was determined with the children in pre-school age of the three communities. Significant evidences (p<0.05) of the high level of acceptability of the food, when comparing the obtained nutritional state at the beginning, half and at the end of the study were detected. Improvement on the health state was observed, when their severe malnutrition normalized.

El objetivo de este estudio fue determinar el impacto nutricional y sensorial de un alimento a base de atún y soya enriquecido con vitaminas y minerales en preescolares de tres comunidades de Chiapas, México. Estudio de intervención nutricional con una cohorte de seis meses. Se determinó el estado nutricional de los niños previo a la alimentación, a la mitad y al final de ésta; la muestra se conformó con los niños que presentaron problemas de desnutrición inicialmente; a los que se les complementó la alimentación durante seis meses. Se determinó el nivel de agrado del producto, con niños de las tres comunidades. Se encontraron evidencias significativas (p<0.05) de un alto nivel de agrado del alimento, al comparar el estado nutricional obtenido al inicio, a la mitad y al final del estudio, se observó mejoras en el estado salud de los niños, al pasar de desnutrición severa o grave a normal.