RESUMO
Emerging technologies, such as focused microwave heating of liquid foods, have been studied to reduce quality losses due to the high temperatures of conventional processing. Besides faster heating, microwaves can also have non-thermal effects on inactivation; however, this is a controversial issue. The objective of this study was to compare conventional and focused microwave heating under similar conditions for the inactivation of two polyphenol oxidases (PPOs): mushroom tyrosinase in buffer and the PPO present in coconut water. Small samples under stirring were treated at temperatures between 50 and 90 °C and three kinetic models were adjusted considering the whole time-temperature history. The Weibull model could best describe inactivation in both heating processes, which was more effective with microwave heating for temperatures over 70 °C. Validation runs show that the model can satisfactorily describe the PPO inactivation. This study contributes for the design of liquid food pasteurization by focused microwave technology.
Assuntos
Catecol Oxidase/química , Pasteurização/métodos , Agaricales/enzimologia , Soluções Tampão , Catecol Oxidase/metabolismo , Cocos/enzimologia , Ativação Enzimática , Indústria de Processamento de Alimentos/métodos , Calefação , Cinética , Micro-Ondas , Modelos Teóricos , Monofenol Mono-Oxigenase/química , Monofenol Mono-Oxigenase/metabolismo , TemperaturaRESUMO
The fragrance gene, betaine aldehyde dehydrogenase 2 (Badh2), has been well studied in many plant species. The objectives of this study were to clone Badh2 and compare the sequences between aromatic and non-aromatic coconuts. The complete coding region was cloned from cDNA of both aromatic and non-aromatic coconuts. The nucleotide sequences were highly homologous to Badh2 genes of other plants. Badh2 consisted of a 1512-bp open reading frame encoding 503 amino acids. A single nucleotide difference between aromatic and non-aromatic coconuts resulted in the conversion of alanine (non-aromatic) to proline (aromatic) at position 442, which was the substrate binding site of BADH2. The ring side chain of proline could destabilize the structure leading to a non-functional enzyme. Badh2 genomic DNA was cloned from exon 1 to 4, and from exon 5 to 15 from the two coconut types, except for intron 4 that was very long. The intron sequences of the two coconut groups were highly homologous. No differences in Badh2 expression were found among the tissues of aromatic coconut or between aromatic and non-aromatic coconuts. The amino acid sequences of BADH2 from coconut and other plants were compared and the genetic relationship was analyzed using MEGA 7.0. The phylogenetic tree reconstructed by the Bayesian information criterion consisted of two distinct groups of monocots and dicots. Among the monocots, coconut (Cocos nucifera) and oil palm (Elaeis guineensis) were the most closely related species. A marker for coconut differentiation was developed from one-base substitution site and could be successfully used.
Assuntos
Betaína-Aldeído Desidrogenase/genética , Cocos/genética , Sequência de Aminoácidos , Cocos/enzimologia , Éxons , Genes de Plantas , Odorantes , Fenótipo , Filogenia , Proteínas de Plantas/genética , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNARESUMO
A novel unmediated hydrogen peroxide biosensor based on the incorporation of fibrous tissue of coconut fruit in carbon paste matrix is presented. Cyclic voltammetry and amperometry were utilized to characterize the main electrochemical parameters and the performance of this new biosensor under different preparation and operation conditions. The resulting H2O2-sensitive biosensors respond rapidly (7 s to attain 90% of the signal), was operated at -0.15 V, presented linear response between 2.0x10(-4) and 3.4x10(-3) mol L(-1), the detection limit was estimated as 4.0x10(-5) mol L(-1). Its operation potential was situated between -0.2 and 0.1 V and the best pH was determined as 5.2. Electrodes containing 5% (w/w) of coconut fiber presented the best signal and their lifetime was extended to 3 months. The apparent Michaelis-Menten constant KM(app) and Vmax were estimated to be 8.90 mmol L(-1) and 6.92 mmol L(-1) microA(-1), respectively. The results obtained for determination of hydrogen peroxide in four pharmaceutical products (antiseptic solution, contact lenses cleaning solution, hair coloring cream and antiseptic dental rinse solution) were in agreement with those obtained by the spectrophotometric method. An additional advantage of these biosensors is the capacity to measure hydrogen peroxide even in samples with relatively low pH. To demonstrate the enzymatic activity of the coconut tissue, a very simple way was created during this work. Coconut fibers were immersed in H2O2 solution between two glass slides. Sequential images were taken to show the rapid generation of O2, attesting the high activity of the enzymes.