How the "Olive Oil Polyphenols" Health Claim Depends on Anthracnose and Olive Fly on Fruits.

Olive anthracnose, caused by Colletotrichum fungi, and the olive fruit fly Bactrocera olea are, respectively, the most important fungal disease and pest affecting olive fruits worldwide, leading to detrimental effects on the yield and quality of fruits and olive oil. This study focuses on the content of hydroxytyrosol (HYT) and its derivatives (the "olive oil polyphenols" health claim) in olive oils extracted from fruits of 'Galega Vulgar' and 'Cobrançosa' cultivars, naturally affected by olive anthracnose and olive fly. The olives, with different damage levels, were harvested from organic rainfed orchards, located in the center of Portugal, at four harvest times over three years. Galega oils extracted from olives with a higher anthracnose and olive fly incidence showed no conformity for the extra virgin olive oil (EVOO) and virgin olive oil (VOO) categories, presenting high acidity and negative sensory notes accompanied by the disappearance of oleacein. Conversely, no sensory defects were observed in Cobrançosa oils, regardless of disease and pest incidence levels, and quality criteria were still in accordance with the EVOO category. The total HYT and tyrosol (TYR) content (>5 mg/20 g) allows for the use of the "olive oil polyphenols" health claim on the label of all the analyzed Cobrançosa olive oils.

Ultrasound Assisted Coextraction of Cornicabra Olives and Thyme to Obtain Flavored Olive Oils.

Flavoring olive oils is a new trend in consumer preferences, and different enrichment techniques can be used. Coextraction of olives with a flavoring agent is an option for obtaining a flavored product without the need for further operations. Moreover, ultrasound (US) assisted extraction is an emergent technology able to increase extractability. Combining US and coextraction, it is possible to obtain new products using different types of olives (e.g., cultivar and ripening stage), ingredient(s) with the greatest flavoring and/or bioactive potential, as well as extraction conditions. In the present study, mastic thyme (Thymus mastichina L.) (TM) and lemon thyme (Thymus x citriodorus) (TC) were used for flavoring Cornicabra oils by coextraction. The coextraction trials were performed by (i) thyme addition to the olives during crushing or malaxation and (ii) US application before malaxation. Several parameters were evaluated in the oil: quality criteria parameters, total phenols, fatty acid composition, chlorophyll pigments, phenolic profile and oxidative stability. US application did not change the phenolic profile of Cornicabra olive oils, while the enrichment of olive oils with phenolic compounds or pigments by coextraction was very dependent on the thyme used. TM enrichment showed an improvement of several new phenolic compounds in the oils, while with TC, fewer new phenols were observed. In turn, in the trials with TC, the extraction of chlorophyll pigments was higher, particularly in crushing coprocessing. Moreover, the oils obtained with US and TM added in the mill or in the malaxator showed lower phenol decrease (59%) than oils flavored with TC (76% decrease) or Cornicabra virgin olive oil (80% decrease) over an 8-month storage period. Multivariate data analysis, considering quality parameters, pigments and phenolic contents, showed that flavored oils were mainly grouped by age.

Continuous Production of Dietetic Structured Lipids Using Crude Acidic Olive Pomace Oils.

Crude olive pomace oil (OPO) is a by-product of olive oil extraction. In this study, low-calorie structured triacylglycerols (TAGs) were produced by acidolysis of crude OPO with medium-chain fatty acids (caprylic, C8:0; capric, C10:0) or interesterification with their ethyl ester forms (C8EE, C10EE). These new TAGs present long-chain fatty acids (L) at position sn-2 and medium-chain fatty acids (M) at positions sn-1,3 (MLM). Crude OPO exhibited a high acidity (12.05-28.75% free fatty acids), and high contents of chlorophylls and oxidation products. Reactions were carried out continuously in a packed-bed bioreactor for 70 h, using sn-1,3 regioselective commercial immobilized lipases (Thermomyces lanuginosus lipase, Lipozyme TL IM; and Rhizomucor miehei lipase, Lipozyme RM IM), in solvent-free media at 40 °C. Lipozyme RM IM presented a higher affinity for C10:0 and C10EE. Lipozyme TL IM preferred C10:0 over C8:0 but C8EE over C10EE. Both biocatalysts showed a high activity and operational stability and were not affected by OPO acidity. The New TAG yields ranged 30-60 and the specific productivity ranged 0.96-1.87 g NewTAG/h.g biocatalyst. Lipozyme RM IM cost is more than seven-fold the Lipozyme TL IM cost. Therefore, using Lipozyme TL IM and crude acidic OPO in a continuous bioreactor will contribute to process sustainability for structured lipid production by lowering the cost of the biocatalyst and avoiding oil refining.

Enrichment of Sunflower Oil with Ultrasound-Assisted Extracted Bioactive Compounds from Crithmum maritimum L.

Crithmum maritimum L., or sea fennel, is an edible halophyte plant, rich in phenolic compounds with antioxidant and antimicrobial activities, that naturally grows in Mediterranean coasts. This study aims to incorporate bioactive compounds extracted from lyophilized Crithmum maritimum to sunflower oil assisted by ultrasounds (UAE), to improve its biological value and oxidative stability. UAE conditions were optimized as a function of time (5-20 min) and lyophilized plant concentration (5-20% m/v). The experiments were dictated by a central composite rotatable matrix. Oxidation products were not influenced by UAE conditions. Acidity, chlorophyll, and carotenoid contents were affected by both factors, while total phenols, flavonoids, and antioxidant activity (FRAP method) only increased with plant concentration. Response surfaces were fitted to these experimental results. Flavonoids were highly related with oil antioxidant activity. No sensory defects were detected in supplemented oil (12.5% m/v plant/5 min UAE). The oxidative stability of this oil was evaluated at 60 °C/12 days. Chlorophylls, phenols, radical scavenging (DPPH), and antioxidant activities decreased over time but were always higher than the values in non-supplemented oil (8.6 and 7-fold with FRAP and DPPH, respectively). C. maritimum presented high amounts of bioactive compounds with antioxidant activity, adequate for sunflower oil supplementation by UAE.

Integrated bioprocess for structured lipids, emulsifiers and biodiesel production using crude acidic olive pomace oils.

Olive pomace oil (OPO), a by-product of olive oil industry, is directly consumed after refining. The novelty of this study consists of the direct use of crude high acidic OPO (3.4-20% acidity) to produce added-value compounds, using sn-1,3-regioselective lipases: (i) low-calorie dietetic structured lipids (SL) containing caprylic (C8:0) or capric (C10:0) acids by acidolysis or interesterification with their ethyl esters, (ii) fatty acid methyl esters (FAME) for biodiesel, and (iii) sn-2 monoacylglycerols (emulsifiers), as by-product of FAME production by methanolysis. Immobilized Rhizomucor miehei lipase showed similar activity in acidolysis and interesterification for SL production (yields: 47.8-53.4%, 7 h, 50℃) and was not affected by OPO acidity. Batch operational stability decreased with OPO acidity, but it was at least three-fold in interesterification that in acidolysis. Complete conversion of OPO into FAME and sn-2 monoacylglycerols was observed after 3 h-transesterification (glycerol stepwise addition) and lipase deactivation was negligeable after 11 cycles.

Argan Oil as a Rich Source of Linoleic Fatty Acid for Dietetic Structured Lipids Production.

Argan oil is rich in long-chain unsaturated fatty acids (FA), mostly oleic and linoleic, and natural antioxidants. This study addresses the production of low-calorie structured lipids by acidolysis reaction, in a solvent-free system, between caprylic (C8:0; system I) or capric (C10:0; system II) acids and argan oil, used as triacylglycerol (TAG) source. Three commercial immobilized lipases were tested: Novozym® 435, Lipozyme® TL IM, and Lipozyme® RM IM. Higher incorporation degree (ID) was achieved when C10:0 was used as acyl donor, for all the lipases tested. Lipozyme® RM IM yielded the highest ID for both systems (28.9 ± 0.05 mol.% C10:0, and 11.4 ± 2.2 mol.% C8:0), being the only catalyst able to incorporate C8:0 under the reaction conditions for biocatalyst screening (molar ratio 2:1 FA/TAG and 55 °C). The optimal conditions for Lipozyme® RM IM in system II were found by response surface methodology (66 °C; molar ratio FA/TAG of 4:1), enabling to reach an ID of 40.9 mol.% of C10:0. Operational stability of Lipozyme® RM IM in system II was also evaluated under optimal conditions, after eight consecutive 24 h-batches, with biocatalyst rehydration between cycles. The biocatalyst presented a half-life time of 103 h.

Co-Processed Olive Oils with Thymus mastichina L.-New Product Optimization.

Olive co-processing consists of the addition of ingredients either in the mill or in the malaxator. This technique allows selecting the type of olives, the ingredients with the greatest flavoring and bioactive potential, and the technological extraction conditions. A new product-a gourmet flavored oil-was developed by co-processing olives with Thymus mastichina L. The trials were performed using overripe fruits with low aroma potential (cv. 'Galega Vulgar'; ripening index 6.4). Experimental conditions were dictated by a central composite rotatable design (CCRD) as a function of thyme (0.4-4.6%, w/w) and water (8.3-19.7%, w/w) contents used in malaxation. A flavored oil was also obtained by adding 2.5% thyme during milling, followed by 14% water addition in the malaxator (central point conditions of CCRD). The chemical characterization of the raw materials, as well as the analysis of the flavored and unflavored oils, were performed (chemical quality criteria, sensory analysis, major fatty acid composition, and phenolic compounds). Considering chemical quality criteria, the flavored oils have the characteristics of "Virgin Olive Oil" (VOO), but they cannot have this classification due to legislation issues. Flavored oils obtained under optimized co-processing conditions (thyme concentrations > 3.5-4.0% and water contents varying from 14 to 18%) presented higher phenolic contents and biologic value than the non-flavored VOO. In flavored oils, thyme flavor was detected with high intensity, while the defect of "wet wood", perceived in VOO, was not detected. The flavored oil, obtained by T. mastichina addition in the mill, showed higher oxidative stability (19.03 h) than the VOO and the co-processed oil with thyme addition in the malaxator (14.07 h), even after six-month storage in the dark (16.6 vs. 10.3 h).

Optimization of Ultrasound-Assisted Extraction of Bioactive Compounds from Pelvetia canaliculata to Sunflower Oil.

In this study, Pelvetia canaliculata L. macroalga, collected from the Atlantic Portuguese coast, was used as a source of bioactive compounds, mostly antioxidants, to incorporate them in sunflower oil with the aim of increasing its biological value and oxidative stability. The lyophilized alga was added to the oil, and ultrasound-assisted extraction (UAE) was performed. Algae concentration and UAE time varied following a central composite rotatable design (CCRD) to optimize extraction conditions. The following parameters were analyzed in the oils: oxidation products, acidity, color, chlorophyll pigments, carotenoids, flavonoids, total phenolic content, antioxidant activity by DPPH (2,2-diphenyl-1-picrylhydrazyl) and FRAP (ferric reducing antioxidant power) assays, and sensory analysis. Extraction conditions did not affect the acidity and the amount of oxidation products in the oil. Chlorophylls and carotenoid contents increased with algae concentration, while flavonoid extraction did not depend on algae content or UAE time. Total phenolics in the oil were highly related only to FRAP antioxidant activity. Storage experiments of supplemented oil (12.5% algae; 20 min UAE) were carried out under accelerated oxidation conditions at 60 °C/12 days. Antioxidant activity (FRAP) of supplemented oil was 6-fold higher than the value of non-supplemented oil. Final samples retained 40% of their initial antioxidant activity. The presence of algae extracts contributed to the increased oxidative stability of sunflower oil.

Phytochemical characterization of phloem in maritime pine and stone pine in three sites in Portugal.

This study analyzes the content and chemical profile of extractives present in the young phloem of mature trees of maritime pine (Pinus pinaster) and stone pine (P. pinea) in three sites in Portugal located in different climatic environments. The cross-sites average of extractives was similar in both pines with 38.5% in P. pinea and 37.7% in P. pinaster phloem. The hydrophilic fraction represented 82% and 70% of P. pinea and P. pinaster total extractives respectively, with large contents of phenolic compounds, flavonoids and tannins, and showed very high oxygen scavenging and reducing ability. Lipophilic extractives were present in higher proportion in P. pinaster phloem than in P. pinea phloem, and showed a large content of resin acids, with the predominance of abietic acid in P. pinaster, and dehydroabietic acid in P. pinea phloems, and of alkanoic acids. P. pinaster and P. pinea have specific defences related to phloem production of resin and phenolic compounds with the ratio phenolic-to-oleoresin compounds higher for P. pinea (4.7 vs 2.3 for P. pinaster) and constant in the three sites. The phytochemical content and composition of the young phloem of P. pinaster and P. pinea showed site differences highlighting the relationship between environment and metabolic production.

Chemical Composition of Cuticular Waxes and Pigments and Morphology of Leaves of Quercus suber Trees of Different Provenance.

The chemical composition of cuticular waxes and pigments and the morphological features of cork oak (Quercus suber) leaves were determined for six samples with seeds of different geographical origins covering the natural distribution of the species. The leaves of all samples exhibited a hard texture and oval shape with a dark green colour on the hairless adaxial surface, while the abaxial surface was lighter, with numerous stomata and densely covered with trichomes in the form of stellate multicellular hairs. The results suggest an adaptive role of leaf features among samples of different provenance and the potential role of such variability in dealing with varying temperatures and rainfall regimes through local adaptation and phenotypic plasticity, as was seen in the trial site, since no significant differences in leaf traits among the various specimens were found, for example, specific leaf area 55.6-67.8 cm2/g, leaf size 4.6-6.8 cm2 and photosynthetic pigment (total chlorophyll, 31.8-40.4 µg/cm2). The leaves showed a substantial cuticular wax layer (154.3-235.1 µg/cm2) composed predominantly of triterpenes and aliphatic compounds (61-72% and 17-23% of the identified compounds, respectively) that contributed to forming a nearly impermeable membrane that helps the plant cope with drought conditions. These characteristics are related to the species and did not differ among trees of different seed origin. The major identified compound was lupeol, indicating that cork oak leaves may be considered as a potential source of this bioactive compound.

Production of low-calorie structured lipids from spent coffee grounds or olive pomace crude oils catalyzed by immobilized lipase in magnetic nanoparticles.

In this study, crude oils extracted from spent coffee grounds (SCG) and olive pomace (OP) were used as raw-material to synthesize low-calorie triacylglycerols, either by acidolysis with capric acid, or by interesterification with ethyl caprate, in solvent-free media, catalyzed by sn-1,3 regioselective lipases. The Rhizopus oryzae lipase (ROL) was immobilized in magnetite nanoparticles (MNP-ROL) and tested as novel biocatalyst. MNP-ROL performance was compared with that of the commercial immobilized Thermomyces lanuginosus lipase (Lipozyme TL IM). For both oils, Lipozyme TL IM preferred interesterification over acidolysis. MNP-ROL catalyzed reactions were faster and acidolysis was preferred with yields of c.a. 50% new triacylglycerols after 3 h acidolysis of OP or SCG oils. MNP-ROL was very stable following the Sadana deactivation model with half-lives of 163 h and 220 h when reused in batch acidolysis and interesterification of OP oil, respectively.

Valorization of lignocellulosic residues from the olive oil industry by production of lignin, glucose and functional sugars.

Spent olive pomace from the two-phase extraction system of virgin olive oil and olive pomace oil, is a major agro-industrial residue. Present study aimed at the valorization of residual olive pomace and stones (seeds) by hydrothermal treatment and enzymatic hydrolysis of glucans. Both residues contain lignin (31.2% and 42.1%), glucans (13.8% and 15.3%) and xylans (18.9% and 20.3%). After hydrothermal pretreatment (130 °C, 30 min; severity factor log R0 = 2.99), 65% and 75% of hemicelluloses (65% of xylan) were hydrolysed into xylo-oligosaccharides in pomace and stones, respectively. Cellulose and lignin were not substantially affected. Three commercial enzyme preparations, Saczyme Yield, Ultimase BWL 40 and Celluclast 1.5 L, were evaluated for saccharification of pomace or stones at three biomass loads (10, 20 and 30%, w/v). Saczyme and Ultimase were active with high solid loads (30%), reaching 80 and 90% of glucan conversion in pomace, and 40 and 55% in stones, respectively, after 5 h.

Lipase-Catalyzed Synthesis of Structured Lipids at Laboratory Scale.

In this chapter, some examples of laboratory protocols to produce functional structured lipids, namely, human milk fat substitutes, dietetic triacylglycerols, and interesterified fat blends with improved nutritional and rheological properties, catalyzed either by immobilized commercial or noncommercial lipase preparations, are presented. In addition to batch synthesis, the continuous production in packed- or fluidized-bed bioreactors is addressed, as well as the evaluation of operational stability of the biocatalysts used (either in batch reuses or in continuous mode).

Application of commercial and non-commercial immobilized lipases for biocatalytic production of ethyl lactate in organic solvents.

This study explores the potential for enhancing the production of ethyl lactate (EL), a green solvent, through enzymatic esterification. Different solvents were compared as organic media for conversion of lactate and ethanol into EL, catalyzed by Novozym 435. Chloroform and hexane were the most effective in low acid concentrations (0.01-0.1M) exhibiting maximum EL yields of 88% and 75% respectively. The yield of EL improved as the solvent's LogP increased up to a value of 2. Non-commercial immobilized biocatalysts consisting heterologous Rhizopous oryzae (rROL) and Candida rugosa (CRL) lipases immobilized on hydrophobic supports were compared to commercial biocatalysts clarifying that Novozym 435 and Lipozyme RM IM could be efficiently applied. Operational stability tests were conducted using Novozym 435, which retained higher activity in chloroform as compared to hexane. Although non-commercial biocatalysts were not competitive in esterification, they exhibited significant activity towards hydrolysis constituting a valuable alternative to higher-cost options.

Influence of enzymes and technology on virgin olive oil composition.

This work aims at presenting the state-of-the-art about the influence of the activity of olive endogenous enzymes, as well as of the application of adjuvants in olive oil technology, discussing their influence on the composition of virgin olive oil, especially in phenols and volatile compounds.

Biodiesel production from crude jatropha oil catalyzed by immobilized lipase/acyltransferase from Candida parapsilosis in aqueous medium.

The lipase/acyltransferase from Candida parapsilosis (CpLIP2) immobilized on two synthetic resins (Accurel MP 1000 and Lewatit VP OC 1600) was used as catalyst for the production of biodiesel (fatty acid methyl esters, FAME) by transesterification of jatropha oil with methanol, in a lipid/aqueous system. The oil was dispersed in a buffer solution (pH 6.5) containing methanol in excess (2M in the biphasic system; molar ratio methanol/acyl chains 2:1). Transesterification was carried out at 30°C, under magnetic stirring, using 10% (w/w) of immobilized enzyme in relation to oil. The maximum FAME yields were attained after 8h reaction time: 80.5% and 93.8%, when CpLIP2 immobilized on Accurel MP 1000 or on Lewatit VP OC 1600 were used, respectively. CpLIP2 on both Accurel MP 1000 and Lewatit VP OC 1600 showed high operational stability along 5 consecutive 8h batches.

Phenolic compounds of 'Galega Vulgar' and 'Cobrançosa' olive oils along early ripening stages.

In this study, the lipophilic and hydrophilic phenol composition of virgin olive oils (VOO) obtained from olives from two of the most important Portuguese cultivars ('Galega Vulgar' and 'Cobrançosa'), harvested at different ripening stages and under two irrigation schemes (rain fed and irrigated), was evaluated. Phenolic alcohols (hydroxytyrosol and tyrosol), phenolic acids and derivatives and flavonoids (luteolin and apigenin), as well as tocopherols were quantified. Lipophilic (>300mgkg(-1)) and hydrophilic phenols (>600mgkg(-1)) were present in high contents in both VOO, for early ripening stages. Gamma-tocopherol content is higher in 'Galega Vulgar' VOO. Total phenols showed a decrease between ripening index 2.5 and 3.5. The dialdehydic form of elenolic acid linked to hydroxytyrosol (3,4-DHPEA-EDA), also known as oleacein, was the major phenolic compound identified in both oils. The concentration of free hydroxytyrosol and tyrosol in both VOO is very low while their esterified derivatives, like 3,4-DHPEA-EDA and p-HPEA-EDA, are much more abundant.

Effect of Ethanol, Sulfur Dioxide and Glucose on the Growth of Wine Spoilage Yeasts Using Response Surface Methodology.

Response surface methodology (RSM) was used to study the effect of three factors, sulfur dioxide, ethanol and glucose, on the growth of wine spoilage yeast species, Zygosaccharomyces bailii, Schizosaccharomyces pombe, Saccharomycodes ludwigii and Saccharomyces cerevisiae. Seventeen central composite rotatable design (CCRD) trials were designed for each test yeast using realistic concentrations of the factors (variables) in premium red wine. Polynomial regression equations were fitted to experimental data points, and the growth inhibitory conditions of these three variables were determined. The overall results showed Sa. ludwigii as the most resistant species growing under high ethanol/free sulfur dioxide concentrations, i.e., 15% (v/v)/20 mg L-1, 14% (v/v)/32 mg L-1 and 12.5% (v/v)/40 mg L-1, whereas other yeasts did not survive under the same levels of ethanol/free sulfur dioxide concentrations. The inhibitory effect of ethanol was primarily observed during longer incubation periods, compared with sulfur dioxide, which showed an immediate effect. In some CCRD trials, Sa. ludwigii and S. cerevisiae showed growth recovery after a short death period under the exposure of 20-32 mg L-1 sulfur dioxide in the presence of 11% (v/v) or more ethanol. However, Sc. pombe and Z. bailii did not show such growth recovery under similar conditions. Up to 10 g L-1 of glucose did not prevent cell death under the sulfur dioxide or ethanol stress. This observation demonstrates that the sugar levels commonly used in wine to sweeten the mouthfeel do not increase wine susceptibility to spoilage yeasts, contrary to the anecdotal evidence.

Supervisão em mediação: uma abordagem sistêmica de apresentação de caso / Group couseling in mediation: a systemic approach for case presentation

O artigo apresenta um modelo de supervisão para apresentação de casos em mediação a partir dos aportes do construcionismo social e o processo U. Este método foi criado, primeiramente, para o contexto do Setor de Mediação do Fórum de Santana. O novo método contribuiu sobremaneira para a capacidade de reflexão e da ampliação do repertório do mediador e, mais especificamente, da sua percepção em relação ao sistema mediador-mediando e comediação.(AU)

This article provides group counseling with the approach of the social constructionism and the U-Process for facilitating the case presentation in mediation. This method was first created for the context of the Forum Santana mediation Sector. The new method contributed greatly to develop the capacity to reflect and to expand mediator’s repertoire, and its perception of mediator-mediating and comediator systems.(AU)

The potential use of lipases in the production of fatty acid derivatives for the food and nutraceutical industries

The potential of lipases (E.C.3.1.1.3.) as biocatalysts for the production of fatty acid derivatives for the food and nutraceutical industries, such as flavouring esters, fatty acid esters of antioxidants and structured lipids, is enormous, mainly due to their high regio- and stereo-selectivities, in addition to the other well-known advantages of enzymatic processes. The replacement of chemical catalysts by lipases presents great benefits in terms of the nutritional properties of the obtained products and environmental care. The reactions performed for the production of these compounds, as well as the best operation conditions, the biocatalysts used, reactor types and operation mode, are addressed in this review.