María Elena Lugo-Sánchez

Comparison of the milk-clotting properties of three plant extracts.

Several proteases from plant sources have been proposed as milk coagulants, however, limited research has been done on their milk-clotting properties. The effect of temperature on the milk-clotting activity of kiwi fruit, melon and ginger extracts was evaluated, as well as the effects of the different extracts on curd properties. Melon extracts showed high milk-clotting activity over a broad temperature range (45-75 °C) while kiwi fruit and ginger extracts showed high activity over a narrower temperature range, with a maximum at 40 and 63 °C, respectively. Curds produced using kiwi extracts had textural properties comparable with those obtained using commercial rennet, while melon extracts produced a fragile gel and low curd yield. The milk-clotting behavior of the three plant extracts was related to the protease specificity present in these extracts. The kiwi proteases displayed chymosin-like properties and thus hold the best potential for use as a milk coagulant in cheese production.

Conformational changes in proteins recovered from jumbo squid (Dosidicus gigas) muscle through pH shift washing treatments.

Conformational and thermal-rheological properties of acidic (APC) and neutral (NPC) protein concentrates were evaluated and compared to those of squid (Dosidicus gigas) muscle proteins (SM). Surface hydrophobicity, sulfhydryl status, secondary structure profile, differential scanning calorimetry and oscillatory dynamic rheology were used to evaluate the effect of treatments on protein properties. Acidic condition during the washing process (APC) promoted structural and conformational changes in the protein present in the concentrate produced. These changes were enhanced during the heat setting of the corresponding sol. Results demonstrate that washing squid muscle under the proposed acidic conditions is a feasible technological alternative for squid-based surimi production improving its yield and gel-forming ability.

Partial characterization of an effluent produced by cooking of Jumbo squid (Dosidicus gigas) mantle muscle

Jumbo squid (Dosidicus gigas) mantle muscle was cooked simulating industrial procedures (95 degrees C x 25 min, 1.2:5 muscle:water ratio). The effluent produced was analyzed for chemical and biochemical oxygen demands (COD and BOD(5), respectively), proximate analysis, flavor-related compounds (free amino acids, nucleotides and carbohydrates) and SDS-PAGE. The COD and BOD(5) exhibited variation among samplings (N=3) (27.4-118.5 g O(2)/L for COD and 11.3-26.7 g O(2)/L for BOD(5)). The effluent consisted of 1% total solids, 75% of which represented crude protein. Sixty percent of the total free amino acid content, which imparts flavor in squid species, corresponded to glutamic acid, serine, glycine, arginine, alanine, leucine and lysine. The nucleotide concentration followed this order, Hx>>ADP>AMP>ATP>IMP>HxR. The variation observed in the present work was probably due to physiological maturity differences among the squid specimens (i.e., juvenile versus mature). Solids present in squid cooking effluent could be recovered and potentially used as flavor ingredients in squid-analog production by the food industry.

Arginine kinase shows nucleoside diphosphate kinase-like activity toward deoxythymidine diphosphate

Arginine kinase (AK) (ATP: L-arginine phosphotransferase, E.C. 2.7.3.3) catalyzes the reversible transfer of ATP γ-phosphate group to L-arginine to synthetize phospho-arginine as a high-energy storage. Previous studies suggest additional roles for AK in cellular processes. Since AK is found only in invertebrates and it is homologous to creatine kinase from vertebrates, the objective of this work was to demonstrate nucleoside diphosphate kinase-like activity for shrimp AK. For this, AK from marine shrimp Litopenaeus vannamei (LvAK) was purified and its activity was assayed for phosphorylation of TDP using ATP as phosphate donor. Moreover, by using high-pressure liquid chromatography (HPLC) the phosphate transfer reaction was followed. Also, LvAK tryptophan fluorescence emission changes were detected by dTDP titration, suggesting that the hydrophobic environment of Trp 221, which is located in the top of the active site, is perturbed upon dTDP binding. The kinetic constants for both substrates Arg and dTDP were calculated by isothermal titration calorimetry (ITC). Besides, docking calculations suggested that dTDP could bind LvAK in the same cavity where ATP bind, and LvAK basic residues (Arg124, 126 and 309) stabilize the dTDP phosphate groups and the pyrimidine base interact with His284 and Ser122. These results suggest that LvAK bind and phosphorylate dTDP being ATP the phosphate donor, thus describing a novel alternate nucleoside diphosphate kinase-like activity for this enzyme.

Optimal Immobilization of Acidic Proteases from Monterey Sardine (Sardinops sagax caeurelea) on Partially Deacetylated Chitin from Shrimp Head Waste

ABSTRACT Response surface methodology was employed to optimize the immobilization yield of acidic proteases from Monterey sardine (Sardinops sagax caeurelea) using partially deacetylated chitin as immobilization support. A rotatable central composite design was applied to evaluate the effects of immobilization conditions such as enzyme loading (X1), immobilization pH (X2), and tripolyphosphate concentration (X3) on the immobilization yield. The analysis of variance revealed that the established model was significant (p < 0.05), and the adjustment of the quadratic model with the experimental data was satisfactory. Under optimal conditions (X1 = 0.05 mg/mL, X2 = 3.16, and X3 = 0.75%), an immobilization yield of 79.1% was achieved; a value that was in accordance with the predicted one.

Relationships between morphometrical properties and the texture of an extrusion-expanded snack made from squid mantle (Dosidicus gigas)

The giant squid (Dosidicus gigas) is a species of commercial interest as a source of protein, and it can be developed into ready-to-eat food products, including expanded extrusion snacks (EES). EES are prepared primarily from starch; however, adding animal protein increases the nutritional contents. The objective of this study was to evaluate the effect of the protein-carbohydrate interactions on the physical and morphological characteristics of an EES made of squid mantle and potato-corn flour. The independent variable was the squid mantle content (40, 60, 80, and 100%) and two controls (01  = 100% potato, 02  = 100% corn). The expansion rate (ER) of the sample is significantly minor (p < .5) when the squid mantle content increases ER = 2.0, 1.8 1.4 to 40, 60, and 80%, respectively. In samples with more protein, crispness and crunchiness were reduced, whereas the hardness increased. Digital imaging analysis indicated that the interaction between protein and starch causes significant morphometric changes to the fractal dimension (2.665-2.739) and lacunarity (0.61-1.29). The results showed that it is possible to incorporate up to 60% squid mantle to prepare EES that possess texture and morphometric characteristics competitive in reported studies with snacks usually incorporating flours, corn, and wheat in the formulations. PRACTICAL APPLICATIONS The giant squid is a very attractive species because its meat has low caloric intake, high protein content, and is an important source of omega 3 fatty acids. Despite the desirable qualities of the squid meat its consumption is very low due to the low diffusion of the properties of its meat, acid, and ammoniacal flavor, rigid texture that requires prolonged cooking times and lack of alternatives of consumption. In Mexico, this type of squid is mainly destined for export as frozen mantle and products with little added value, which generates little economic benefit. Therefore, the results of this research may be of interest to the squid processing industry, which demands new forms of consumption of this marine species to increase their commercialization and added value.

Partial Characterization of AMP Deaminase from Jumbo Squid (Dosidicus gigas) Mantle

AMP deaminase enzyme showed an optimum temperature at 35°C. It was relatively stable between 30 and 40°C but extremely unstable at higher temperatures. The optimum pH was 6.0 and the enzyme presented an excellent stability with pH between 5.5 and 6.5. The K+ ion acted as a negative effector when used in concentrations higher than 50 mM. The nucleotide ATP acted as a positive effector in concentrations exceeding 2 mM, while ADP was a positive effector between 0 and 2 mM. These results indicated that AMP deaminase is an important enzyme in Dosidicus gigas post-mortem biochemistry, involving the rapid degradation of AMP during iced storage of jumbo squid mantle.