Maricarmen Quirasco

Development of a liquid nutritional supplement using a Sesamum indicum L. protein isolate

This work presents some functional properties and the potential use of a new protein isolate from sesame seed flour, Sesaprot® (SP), as protein source in a liquid nutritional supplement. It was compared to a commercial soybean isolate and results showed that its emulsifying properties are better than those of its soybean counterpart. Lysine content is lower than FAO recommendations for children, but is adequate for adults. Other essential amino acids, however, are present in adequate amounts for any kind of consumer. Osmolality, pH and emulsion stability of an experimental formulation were similar to those of commercial beverages. A consumer sensory test indicated that the product was preferred to one prepared with a soybean isolate and to a commercial brand that was assayed.

Real-time and conventional PCR detection of Liberty Link rice varieties and transgenic soy in rice sampled in the Mexican and American retail markets.

Samples of rice from Mexican and USA retail stores were analyzed for the presence of transgenic (GM) events using real-time PCR. In screening for the CaMV35S promoter sequence (35SP), positive results were found in 49 and 35% of the Mexican and American samples, respectively. In further investigations in Mexican samples, 43% were positive for P35S::bar, with two above the quantifiable limit; these were 0.07% and 0.05% GMO. Fourteen out of the sixteen positive samples were labeled as imported from the USA. In testing samples bought in American retail shops, 24% showed positive results, all below the quantifiable range. It could be deduced that P35S::bar positive samples were Liberty Link® (LL) rice. In distinguishing between LL601 and LL62, end-point PCR was used, corroborating the P35S::bar amplicon length difference of these events. LL62 was found in one rice sample purchased in Mexico and two in the USA samples. Its presence was verified with the 35S terminator sequence. All other LL positive samples contained LL601. None of the samples analyzed showed the presence of Bt63 rice. The LL rice varieties found have been identified as not being commercially cultivated, and so their presence requires further investigation. 35SP was also present in samples which did not have any LL rice. Maize sequences could not be detected in any of the samples; however, soybean DNA was found in Mexican and USA rice samples. The Roundup Ready® trait was detected in trace amounts in 16 and 6% of the rice samples bought in Mexico and the USA, respectively. Real-time PCR was shown to be the method of choice for the sensitive and rapid screening of commodities and retail samples for the detection of GM and other contamination.

Emulsifying Properties of Chemically Deamidated Corn (Zea Mays) Gluten Meal

Corn gluten meal is a by-product of starch production that is readily available. Corn protein isolates have limited applications due to their hydrophobic nature, low solubility and limited functionality as emulsifiers. In this study, a mild acidic treatment of corn gluten meal was performed in order to achieve deamidation of asparagine and glutamine residues and modify the interfacial behavior of this byproduct. A 0.1 N HCl treatment for 6 h at 70 °C rendered a deamidation degree of 20.4%, which increased the emulsification activity index of corn gluten meal from 6.8 to 16.8 m2/g protein, with a remarkable increase in emulsion stability from 0 to 90.6% oil retention. Proteins participating in the emulsion were separated by SDS-PAGE and the main polypeptides were identified as alpha and beta-zeins. After deamidation, protein dissociation and unfolding due to the obtained negative charges resulted in enhanced functionality.

Cloning and Characterization of a Novel N-acetylglucosaminidase (AtlD) from Enterococcus faecalis

The atlD gene from an Enterococcus faecalis strain isolated from a Mexican artisanal cheese was cloned, sequenced and expressed in Escherichia coli in order to perform a biochemical characterization. A partial amino acid sequence of the heterologous protein was obtained by LC-MS/MS, and it corresponded to a novel peptidoglycan hydrolase designated AtlD. Its molecular mass was 62–75 kDa, as determined by SDS-PAGE, zymography, Western blot, and exclusion chromatography. Electrofocusing rendered an isoelectric point (pI) of 4.8. It exhibited N-acetylglucosaminidase activity, with an optimal pH and temperature between 6–7 and 50°C, respectively. It retained 85% activity with NaCl at 1,000 mM, but it was susceptible to divalent ions, particularly Zn2+. It showed antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, and enterococcal strains of clinical origin. Due to the fact that it showed activity versus pathogenic bacteria, and because of its capabilities under ionic strength, temperature, and pH values present in food matrices, it could be applied as an additive in the food industry. This study will aid in the design of new antibacterial agents of natural origin to combat food-borne diseases, and it could be used as an industrial or hospital hygiene agent as well.

Enhancement of the antibacterial activity of an E. faecalis strain by the heterologous expression of enterocin A

The genus Enterococcus occurs as native microbiota of fermented products due to its broad environmental distribution and its resistance to salt concentrations. Enterococcus faecalis F, a non-pathogenic strain isolated from a ripened cheese, has demonstrated useful enzymatic capabilities, a probiotic behavior and antibacterial activity against some food-borne pathogens, mainly due to peptidoglycan hydrolase activity. Its use as a natural pathogen-control agent could be further enhanced through the production of a bacteriocin, e.g. Enterocin A, because of its remarkable antilisterial activity. In this work, a markerless allelic insertion method was used to obtain an enterococcal strain capable of producing a functional enterocin. Agar diffusion tests showed that the recombinant strain was active against Staphylococcus aureus, Listeria monocytogenes and the pathogenic strain E. faecalis V583. When grown in liquid culture together with L. monocytogenes, it attained a two-log reduction of the pathogen counts in lesser time relative to the native strain. Because the DNA construction is integrated into the chromosome, the improved strain avoids the use of antibiotics as selective pressure; besides, it does not require an inductor because of the inclusion of a constitutive promoter in the construction. Its technological and antibacterial capabilities make the improved E. faecalis strain a potential culture for use in the food industry.