Progress in nanostructure understanding of edible crystalline fats and their application in nano-delivery systems: Cocoa butter as a model.

Nucleation and crystal growth of edible fats at the nanoscale have received little attention due to analytical limitations. A key factor to modify the properties of edible fats is nanostructure understanding. Progress within the last years will be presented, including analytical techniques used to investigate fat crystallization. Cocoa butter has been the subject of several works due to its properties and its high impact on chocolate manufacturing. Moreover, this vegetable fat has been used as the solid lipid component in nano delivery systems. Since nanoplatelet is the smallest unit in crystalline fats, and the nanoscale is influenced by supersaturation, temperature, shear fields, and surfactants, nanostructure engineering is possible. On its part, cocoa butter has been included in innovative delivery systems along the last years. This review will highlight main results and challenges on these topics.

Sodium carbonate versus borate buffer for lactase quenching, laboratory work.

A laboratory exercise for undergraduate advanced students of enzymology and biocatalysis is presented. Since enzyme assays can be quenched or continuous, this experiment compares the performance of two quenching agents for lactase, in a continuous setup. Enzymatic activity of ß-galactosidase (Aspergillus oryzae) was determined based on the release of 4-nitrophenol from 4-nitrophenyl ß-D-galactopyranoside using a microplate reader. Sodium carbonate and borate buffer were tested as quenching agents, and experimental control was the unstopped assay. Based on released 4-nitrophenol, enzyme activity, and rate constant k, the students could assess the performance of each termination agent. The experiment promotes disciplinary and transversal competencies, including research-based learning, critical thinking, and introduce the students to high-throughput techniques that are common in the research and development environment.

Protein A chromatography: Challenges and progress in the purification of monoclonal antibodies.

Antibodies for therapeutic use are being continuously approved and their demand has been steadily growing. As known, the golden standard for monoclonal antibody (mAb) purification is Protein A affinity chromatography, a technology that has gained high interest because of its great performance and capabilities. The main concerns are the elevated resins costs and their limited lifetime compared to other resins (e.g. ion exchange chromatography). Great efforts have been carried out to improve purification conditions, such as resin characterization and designing alkali/acid stable resins with a longer lifetime. Modification of Protein A ligands and alternative formats such as monoliths membranes and microshperes have been tested to increase the purification performance. New technology has been proposed to improve the large-scale separation; in addition, alternative ligands have been suggested to capture mAbs instead of Protein A ligand; however, most of the information is locked by pharmaceutical companies. This mini review summarizes and describes the advances, results, and impact on the Protein A chromatography purification processing.

Rapid physicochemical characterization of innovative fucoidan/fructan powders by ATR-FTIR.

Functional food has been highly demanded lately because of its benefits in counteracting diseases. Fucoidan and agave fructan are ingredients that enhance the growth of beneficial bacteria in the gut (prebiotics). This mixture has great potential to develop innovative products but it has never been explored before. Because of fucoidan is more expensive than agave fructan, the innovative proposed mixture is vulnerable to adulteration. This research was aimed to assess the accuracy of Fourier transform infrared spectroscopy with attenuated total reflectance (ATR-FTIR) coupled with chemometrics to identify and predict concentration of both polysaccharides in powder mixtures (0-100%). Absorption bands at 1240-1255 and 836-840 cm-1 were attributed to fucoidan and a strong peak at ~ 936 cm-1 confirmed the fructan presence. Peak areas were best fitted into linear models ( R adj 2  ≥ 0.92, RMSE ≤ 3.54%). This achievement may be useful to certificate ingredients contained in fucoidan-fructan mixtures, preventing adulteration.

Temperature model for process impact non-uniformity in genipin recovery by high pressure processing.

A model for the process impact temperature non-uniformity during high pressure processing (HPP) of genipap fruit purees was found during genipin recovery. Purees were subjected to HPP (130-530 MPa) under quasi-isobaric non-isothermal conditions (15 min; 0, 4.6 and 9.3mg pectinases/g fruit). Genipin and protein concentration was determined, and pH was measured. Polygalacturonase activity was quantified indirectly by protein content (mg/g fruit). First order kinetics described temperature changes (0-4 min). Polygalacturonase was activated at 130 MPa, inactivated reversibly at 330 MPa and activated again at 530 MPa. Enzyme reaction rate constant (k) was placed in the 0-4 min model and temperature from 2 to 15 min was described. Protein content and pH characterization in terms of decimal reduction time improved highly the 2-15 min model. Since temperature changes were modeled, more insight of its behavior in an HPP reactor was obtained, avoiding uniformity assumptions, making easier the industrial scale HPP implementation.

Environmental friendly cold-mechanical/sonic enzymatic assisted extraction of genipin from genipap (Genipa americana).

An efficient cold-mechanical/sonic-assisted extraction technique was developed for extraction of genipin from genipap (Genipa americana) peel. Ultrasound assisted extraction (285 W, 24 kHz) was performed at 5, 10 and 15 °C for 5, 10 and 15 min. After cold-extraction, genipin was separated from pectin and proteins by aid of fungal pectinesterase. The maximum yield of non-cross-linked genipin was 7.85±0.33 mg/g, at 10 °C for 15 min by means of ultrasound extraction. The protein amount in extracts decreased in all samples. If mechanical process is combined with ultrasound assisted extraction the yield is increased by 8 times after the pectinesterase-assisted polyelectrolyte complex formation between pectic polysaccharides and proteins, avoiding the typical cross-linking of genipin. This novel process is viable to obtain non-cross-linked genipin, to be used as a natural colorant and cross-linker in the food and biotechnological industries.