Effect of modified atmosphere applied to minimally processed radicchio (Cichorium intybus L.) submitted to different sanitizing treatments

Stability of minimally processed radicchio (Cichorium intybus L.) was evaluated under modified atmosphere (2% O2, 5% CO2, and 93% N2) on 3, 5, 7 and 10 days of storage at 5°C. The samples were hygienized in sodium hypochlorite or hydrogen peroxide solutions to identify the most effective sanitizing solution to remove microorganisms. Microbiological analysis was conducted to identify the presence of coliforms at 35°C and 45°C, mesophilic microorganisms, and yeast and mold. Physicochemical analyses of mass loss, pH, soluble solids, and total acidity were conducted. The color measurements were performed using a Portable Colorimeter model CR-400. The antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic methods. The sensory evaluation was carried out using a hedonic scale to test overall acceptance of the samples during storage. The sodium hypochlorite (150 mg.L-1) solution provided greater safety to the final product. The values of pH ranged from 6.17 to 6.25, total acidity from 0.405 to 0.435%, soluble solids from 0.5 to 0.6 °Brix, mass loss from 1.7 to 7.2%, and chlorophyll from 1.068 to 0.854 mg/100g. The antioxidant activity of radicchio did not show significant changes during the first 3 days of storage. The overall acceptance of the sample stored in the sealed package without modified atmosphere was 70%, while the fresh sample was obtained 77% of approval. Although the samples packaged under modified atmosphere had a higher acceptance score, the samples in sealed packages had satisfactory results during the nine days of storage. The use of modified atmosphere, combined with cooling and good manufacturing practices, was sufficient to prolong the life of minimally processed radicchio, Folha Larga cultivar, for up to ten days of storage.

Forced-air, vacuum, and hydro precooling of cauliflower (Brassica oleracea L. var. botrytis cv. Freemont): Part II. Determination of quality parameters during storage

Cauliflower heads, which were precooled using four different methods including vacuum, forced-air, and high and low flow hydro precooling, were stored under controlled atmosphere and room conditions. Controlled atmosphere conditions (CA) were as follows: 1°C temperature, 90 ± 5% relative humidity, and 0:21 [(%CO2:%O2) – (0:21) control] atmosphere composition. Room conditions (RC) were: 22±1°C temperature and 55-60% humidity. Various quality parameters of the cauliflower heads were assessed during storage (days 0, 7, 14, 21, 28, and 35) under controlled atmosphere and room conditions (days 0, 5, and 10). During storage, weight loss, deterioration rate, overall sensory quality score, hardness, and colour (L, a, b, C and α) were evaluated. In the present study, the strength and quality parameters of cauliflower under CA and RC conditions were obtained. Vacuum precooling was found to be most suitable method before cauliflower was submitted to cold storage and sent to market. Furthermore, the storage of cauliflower without precooling resulted in a significant decrease in quality parameters.

Inibição do desenvolvimento in vitro de embriões de Coffea por cafeína exógena

A cafeína, alcalóide conhecido como 1,3,7-trimetilxantina, é encontrada em sementes quiescentes de cafeeiro, perfazendo um total de 1,1 a 1,7% em Coffea arabica L. e 2 a 3% em Coffea canephora Pierre, localizada em sua grande maioria no endosperma, na forma livre no citoplasma das células ou complexada com ácidos clorogênicos. Com função fisiológica em plantas ainda não totalmente esclarecida, a cafeína causa efeito alelopático, seja inibindo a germinação de várias espécies, seja como anti-herbívoro ou como agente pesticida natural. A lenta germinação de sementes de café ainda não foi esclarecida e várias causas são apontadas, como presença do endocarpo, baixa absorção de água e O2, presença de inibidores naturais e balanço hormonal. Embora sugeridos, estudos sobre a inibição de sementes de cafeeiro por ação da cafeína endógena e ou exógena são escassos. Assim, o presente trabalho teve como objetivo estudar o efeito da cafeína exógena sobre a germinação e o desenvolvimento de embriões de Coffea arabica L. e de Coffea canephora Pierre. O experimento foi realizado utilizando-se frutos no estádio cereja das cultivares Rubi e Apoatã IAC-2258. Após desinfestação dos frutos por 30 minutos de imersão em hipoclorito de sódio (2% i.a.) e lavagem por três vezes em água destilada e autoclavada, os embriões foram retirados e inoculados, de modo asséptico, em placas de petri com meio MS 50%, acrescido de sacarose e suplementado com diferentes concentrações de cafeína (0,00; 0,05; 0,10; 0,15; 0,20; 0,25; 0,30 e 0,40%). Os embriões foram mantidos em sala de crescimento a 27 ± 2ºC e densidade de fluxo de fótons de 13µmol.m-2.s-1, durante 23 dias, quando foram avaliados comprimento da parte aérea, comprimento de raiz e massa fresca das plântulas. Cinco dias após o cultivo, foram avaliadas a porcentagem de emissão de radículas e cotilédones, computando-se os embriões com cotilédones abertos e radículas expandidas. O delineamento experimental utilizado foi inteiramente casualizado com seis repetições por tratamento, sendo cada repetição constituída por cinco embriões. Concluiu-se que a germinação e o desenvolvimento in vitro de embriões de Coffea arabica L. e Coffea canephora Pierre são afetados por cafeína exógena. O efeito detrimental da cafeína exógena em embriões de Coffea é maior nas radículas do que nos cotilédones. Embriões de Coffea arabica L. são mais sensíveis aos efeitos negativos da cafeína exógena do que embriões de Coffea canephora Pierre. A cafeína pode contribuir para a lenta germinação de sementes e o lento desenvolvimento de plântulas de cafeeiro.

Alternative electron transfer routes involved in photoprotection of cyanobacteria

In oxygenic photosynthesis, the highly oxidizing reactions of water splitting produce reactive oxygen species (ROS) and other radicals that could damage the photosynthetic apparatus and affect cell viability. Under particular environmental conditions, more electrons are produced in water oxidation than can be harmlessly used by photochemical processes for the reduction of metabolic electron sinks. In these circumstances, the excess of electrons can be delivered, for instance, to O2, resulting in the production of ROS. To prevent detrimental reactions, a diversified assortment of photoprotection mechanisms has evolved in oxygenic photosynthetic organisms. In this thesis, I focus on the role of alternative electron transfer routes in photoprotection of the cyanobacterium Synechocystis sp. PCC 6803. Firstly, I discovered a novel subunit of the NDH-1 complex, NdhS, which is necessary for cyclic electron transfer around Photosystem I, and provides tolerance to high light intensities. Cyclic electron transfer is important in modulating the ATP/NADPH ratio under stressful environmental conditions. The NdhS subunit is conserved in many oxygenic phototrophs, such as cyanobacteria and higher plants. NdhS has been shown to link linear electron transfer to cyclic electron transfer by forming a bridge for electrons accumulating in the Ferredoxin pool to reach the NDH-1 complexes. Secondly, I thoroughly investigated the role of the entire flv4-2 operon in the photoprotection of Photosystem II under air level CO2 conditions and varying light intensities. The operon encodes three proteins: two flavodiiron proteins Flv2 and Flv4 and a small Sll0218 protein. Flv2 and Flv4 are involved in a novel electron transport pathway diverting electrons from the QB pocket of Photosystem II to electron acceptors, which still remain unknown. In my work, it is shown that the flv4-2 operon-encoded proteins safeguard Photosystem II activity by sequestering electrons and maintaining the oxidized state of the PQ pool. Further, Flv2/Flv4 was shown to boost Photosystem II activity by accelerating forward electron flow, triggered by an increased redox potential of QB. The Sll0218 protein was shown to be differentially regulated as compared to Flv2 and Flv4. Sll0218 appeared to be essential for Photosystem II accumulation and was assigned a stabilizing role for Photosystem II assembly/repair. It was also shown to be responsible for optimized light-harvesting. Thus, Sll0218 and Flv2/Flv4 cooperate to protect and enhance Photosystem II activity. Sll0218 ensures an increased number of active Photosystem II centers that efficiently capture light energy from antennae, whilst the Flv2/Flv4 heterodimer provides a higher electron sink availability, in turn, promoting a safer and enhanced activity of Photosystem II. This intertwined function was shown to result in lowered singlet oxygen production. The flv4-2 operon-encoded photoprotective mechanism disperses excess excitation pressure in a complimentary manner with the Orange Carotenoid Protein-mediated non-photochemical quenching. Bioinformatics analyses provided evidence for the loss of the flv4-2 operon in the genomes of cyanobacteria that have developed a stress inducible D1 form. However, the occurrence of various mechanisms, which dissipate excitation pressure at the acceptor side of Photosystem II was revealed in evolutionarily distant clades of organisms, i.e. cyanobacteria, algae and plants.

Monooxygenase Diversity and Oxygen Activation within Polyketide Antibiotic Biosynthesis

Molecular oxygen (O2) is a key component in cellular respiration and aerobic life. Through the redox potential of O2, the amount of free energy available to organisms that utilize it is greatly increased. Yet, due to the nature of the O2 electron configuration, it is non-reactive to most organic molecules in the ground state. For O2 to react with most organic compounds it must be activated. By activating O2, oxygenases can catalyze reactions involving oxygen incorporation into organic compounds. The oxygen activation mechanisms employed by many oxygenases to have been studied, and they often include transition metals and selected organic compounds. Despite the diversity of mechanisms for O2 activation explored in this thesis, all of the monooxygenases studied in the experimental part activate O2 through a transient carbanion intermediate. One of these enzymes is the small cofactorless monooxygenase SnoaB. Cofactorless monooxygenases are unusual oxygenases that require neither transition metals nor cofactors to activate oxygen. Based on our biochemical characterization and the crystal structure of this enzyme, the mechanism most likely employed by SnoaB relies on a carbanion intermediate to activate oxygen, which is consistent with the proposed substrate-assisted mechanism for this family of enzymes. From the studies conducted on the two-component system AlnT and AlnH, both the functions of the NADH-dependent flavin reductase, AlnH, and the reduced flavin dependent monooxygenase, AlnT, were confirmed. The unusual regiochemistry proposed for AlnT was also confirmed on the basis of the structure of a reaction product. The mechanism of AlnT, as with other flavin-dependent monooxygenases, is likely to involve a caged radical pair consisting of a superoxide anion and a neutral flavin radical formed from an initial carbanion intermediate. In the studies concerning the engineering of the S-adenosyl-L-methionine (SAM) dependent 4-O-methylase DnrK and the homologous atypical 10-hydroxylase RdmB, our data suggest that an initial decarboxylation of the substrate is catalyzed by both of these enzymes, which results in the generation of a carbanion intermediate. This intermediate is not essential for the 4-O-methylation reaction, but it is important for the 10-hydroxylation reaction, since it enables substrate-assisted activation of molecular oxygen involving a single electron transfer to O2 from a carbanion intermediate. The only role for SAM in the hydroxylation reaction is likely to be stabilization of the carbanion through the positive charge of the cofactor. Based on the DnrK variant crystal structure and the characterizations of several DnrK variants, the insertion of a single amino acid in DnrK (S297) is sufficient for gaining a hydroxylation function, which is likely caused by carbanion stabilization through active site solvent restriction. Despite large differences in the three-dimensional structures of the oxygenases and the potential for multiple oxygen activation mechanisms, all the enzymes in my studies rely on carbanion intermediates to activate oxygen from either flavins or their substrates. This thesis provides interesting examples of divergent evolution and the prevalence of carbanion intermediates within polyketide biosynthesis. This mechanism appears to be recurrent in aromatic polyketide biosynthesis and may reflect the acidic nature of these compounds, propensity towards hydrogen bonding and their ability to delocalize π-electrons.

Revue de l'Institut oriental / Fondateur l'abbé L. Gruel

Variante(s) de titre : Revue de l'Institut oriental et colonial

Revue de l'Institut oriental / Fondateur l'abbé L. Gruel

1913.