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Fatty acids are produced by eukaryotes like baker’s yeast Saccharomyces cerevisiae mainly using a large multifunctional type I fatty acid synthase (FASI) where seven catalytic steps and a carrier domain are shared between one or two protein subunits. While this system may offer efficiency in catalysis, only a narrow range of fatty acids are produced. Prokaryotes, chloroplasts and mitochondria rely instead on a FAS type II (FASII) where each catalytic step is carried out by a monofunctional enzyme encoded by a separate gene. FASII is more flexible and capable of producing a wider range of fatty acid structures, such as the direct production of unsaturated fatty acids. An efficient FASII in the preferred industrial organism S. cerevisiae could provide a platform for developing sustainable production of specialized fatty acids. We functionally replaced either yeast FASI genes (FAS1 or FAS2) with a FASII consisting of nine genes from Escherichia coli (acpP, acpS and fab -A, -B, -D, -F, -G, -H, -Z) as well as three from Arabidopsis thaliana (MOD1, FATA1 and FATB). The genes were expressed from an autonomously replicating multicopy vector assembled using the Yeast Pathway Kit for in-vivo assembly in yeast. Two rounds of adaptation led to a strain with a maximum growth rate (μmax) of 0.19 h− 1 without exogenous fatty acids, twice the growth rate previously reported for a comparable strain. Additional copies of the MOD1 or fabH genes resulted in cultures with higher final cell densities and three times higher lipid content compared to the control., This work was supported by the Fundação para a Ciência e Tecnologia Portugal (FCT) through Project FatVal POCI-01- 0145-FEDER-032506; CBMA was supported by the strategic program UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through the COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI)., info:eu-repo/semantics/publishedVersion

Principal Component Analysis (PCA) transforms the original variable into new ones called principal components (PC). These PC´s are calculated attributing a coefficient for each original variables proportional to their contribution into this transformation in order to maximize the variances of the first few components [1]. The main objective is to reduce the dimensionality, while keeping the contribution of all initial variables in order to provide a visual pattern recognition [2]. PCA biplot graphs with both scores and loadings provide information on the influence of each variable on a given sample. The hierarchical clustering was also employed in order to highlight the similarities among samples. The following variables were determined through ethanolic extracts of apple and pear varieties from the Alcobaça region (Portugal) using spectrophotometric analysis: DPPH radical scavenging, -carotene bleaching, total phenolic content, total flavonoid content, and fructose content. The results show that a very high correlation exists among the variables antioxidant capacity through DPPH, total phenolics content and total flavonoids, while the fructose content shows independent behavior in relation to the other ones. The graph of scores for the first two PCs, which explain 90% of variance, shows three different clusters with different apple and pear species. It can be concluded from these results that the fructose content allows to separate apples and pears while antioxidant capacity through DPPH, total phenolics content and total flavonoids content can be used to separate different cultivars of each fruit. This study shows that multivariate analysis, with special focus on PCA, can be a valuable tool for the separation of different fruit species and their cultivars highlighting the similarities and differences among them., This study was carried out in the frame of the clabel+ project: Innovative Natural, Nutritious and Consumer Oriented “Clean Label” Foods with the reference POCI-01-0247-FEDER-046080 financed by the Competitiveness and Internationalization Thematic Operational Programme (PO CI), under the COMPETE2020, PORTUGAL2020 Partnership Agreement, through the co-financing of European Regional Development Fund (FEDER). J. D. Teixeira would like to thank to clabel+ project for his fellowship (28/2021/BI). The authors thank Patricia Vicente for her support in the field work. The work was supported by UIDB/00211/2020 with funding from FCT/MCTES through national funds. C. Almeida also acknowledges the financial support by LA/P/0045/2020 (ALiCE), UIDB/00511/2020 and UIDP/00511/2020 (LEPABE), funded by national funds through FCT/MCTES (PIDDAC)., info:eu-repo/semantics/publishedVersion

Phenolic compounds are believed to have antioxidant properties, and therefore many studies have been conducted in order to search for the presence of such compounds in food matrices and the effects that they cause [1, 2]. Some of the most common sources of phenolic compounds in human diet are fruit and fruit based products, particularly apples [3]. An Ultra-High Performance Liquid Chromatography combined with Time-of-Flight Mass Spectrometry (UHPLC-ToF-MS) method was developed for the determination of individual phenolics in the pulp and by-products of regional and commercial cultivars of apples from Alcobaça region (Portugal). The analytical method was evaluated regarding linearity, limit of detection, limit of quantification and accuracy, showing its suitability for the quantification of phenolic compounds. Epicatechin, quercetin-3-b-d-glucoside, quercetin, chlorogenic acid and 4-hydroxybenzoic acid were the main phenolic compounds found in all portions of apples, namely: peels, seeds, and pulp (mesocarp). Apple peels presented the highest content of phenolic compounds. For example, in the Noiva variety, peels had the highest concentration of epicatechin (80.69 ± 0.59 g/g), followed by seeds (16.36 ± 0.48 g/g) and lower levels were found in the pulp (3.74 ± 0.08 g/g). Phloridzin, one of the most recognized apple polyphenols, was quantified in all apples, however, different cultivars have different levels of this phenolic compound. In peels, the level of phloridzin ranges between 10.8 g/g in the Bonita variety to 0.57 mg/g in the Fujion variety. Some cultivars of apples showed very high levels of phenolic compounds, associated with their putative benefits for Human Health. Moreover, their by-products showed great potential as source of natural antioxidants., This study was carried out in the frame of the clabel+ project: Innovative Natural, Nutritious and Consumer Oriented “Clean Label” Foods with the reference POCI-01-0247-FEDER-046080 financed by the Competitiveness and Internationalization Thematic Operational Programme (PO CI), under the COMPETE2020, PORTUGAL2020 Partnership Agreement, through the co-financing of European Regional Development Fund (FEDER) and of the IDfoods project: Food System of The Future - Research and Development in Sustainable Agrifood Systems and Healthy Nutrition with the reference POCI-01-0247- FEDER-039364 supported financed by the Competitiveness and Internationalization Operational Program (POCI), within the scope of COMPETE2020, Partnership Agreement PORTUGAL 2020, co-financed by the European Regional Development Fund (ERDF). The work was supported by UIDB/00211/2020 with funding from FCT/MCTES through national funds. J.D.T. would like to thank to clabel+ project for his fellowship (28/2021/BI). C. Almeida also acknowledges the financial support by LA/P/0045/2020 (ALiCE), UIDB/00511/2020 and UIDP/00511/2020 (LEPABE), funded by national funds through FCT/MCTES (PIDDAC)., info:eu-repo/semantics/publishedVersion

Alcobaça is a region located in the centre of Portugal known by its unique edaphoclimatic conditions that make it the largest apple production region in the country. The Maca de Alcobaca is one of the 1257 products registered as Protected Geographical Origin (PGI) by the eAmbrosia database of the EU [1]. This fruit is widely appreciated worldwide, not only because of the organoleptic properties, but also because of their antioxidant properties and association with lower risk of cardiovascular diseases [2-3]. The objective of this work was to determine the antioxidant properties as well as estimate the fructose content of 30 commercial cultivars produced in the Alcobaça region. The antioxidant properties were evaluated through antioxidant capacity tests (DPPH radical scavenging and -carotene bleaching). Moreover total phenolic content and total flavonoids were also determined. The fructose content was assessed as per Ashwell [4]. Overall, the by-products of the commercial cultivars of apple presented a higher antioxidant capacity than the pulp. The Fujion cultivar, for instance, presented a antioxidant capacity of 76.4 g Trolox equivalents (TE)/g of fresh fruit on the peels, comparatively to 53.7 g TE/g of fresh fruit on the seeds and 22.1 g TE/g of fresh fruit in the pulp. These results show that the by-products of the studied commercial cultivars of the studied apples fruit can be used as a valuable source of natural antioxidants, avoiding their waste and guaranteeing their valorisation for instance, by food industry, in a circular economy concept., This study was carried out in the frame of the clabel+ project: Innovative Natural, Nutritious and Consumer Oriented “Clean Label” Foods with the reference POCI-01-0247-FEDER-046080 financed by the Competitiveness and Internationalization Thematic Operational Programme (PO CI), under the COMPETE2020, PORTUGAL2020 Partnership Agreement, through the co-financing of European Regional Development Fund (FEDER), and of the IDfoods project: Food System of The Future - Research and Development in Sustainable Agrifood Systems and Healthy Nutrition with the reference POCI-01-0247- FEDER-039364 supported financed by the Competitiveness and Internationalization Operational Program (POCI), within the scope of COMPETE2020, Partnership Agreement PORTUGAL 2020, co-financed by the European Regional Development Fund (ERDF).. The work was supported by UIDB/00211/2020 with funding from FCT/MCTES through national funds. J.D.T. would like to thank to clabel+ project for his fellowship (28/2021/BI). C. Almeida also acknowledges the financial support by LA/P/0045/2020 (ALiCE), UIDB/00511/2020 and UIDP/00511/2020 (LEPABE), funded by national funds through FCT/MCTES (PIDDAC)., info:eu-repo/semantics/publishedVersion

Pear (Pyrus communis L.) is a fruit of great interest because their consumption offers nutritional as well as medicinal advantages [1]. The most well-known and studied properties associated with pears are antioxidant and anti-inflammatory properties [2] these are associated with the presence of phenolic compounds, mainly in the peels but also, in smaller amounts, in its mesocarp (pulp) [3]. In this work, an Ultra-High Performance Liquid Chromatography combined with Time-of-Flight Mass Spectrometry (UHPLC-ToF-MS) method was optimized and validated for the determination of individual phenolics in the pulp and by-products of fruits. The main phenolic compounds found in the studied regional pear cultivars (6 cultivars from Alcobaça region) were catechin, epicatechin, caffeic acid, chlorogenic acid, and vanillic acid. The pear by-products (peels and seeds) presented higher phenolic content than mesocarp, mainly peels samples. One example of this is the Carapinheira Roxa cultivar, wherein the peels had the highest concentration of chlorogenic acid (26.26 ± 0.65 g/g) and lower levels were found in the seeds (3.11 ± 0.19 g/g) and in the pulp (4.27 ± 0.03 g/g). Other very well recognized pear phenolics, were quantified in all the pear cultivars, at different levels. For example, in the peels portion, the level of epicatechin varied between 0.19 g/g in the Torres Novas cultivar and 37.30 g/g in the Lambe-os-Dedos cultivar. The studied regional pear cultivars, in particular their by-products, presented high levels of phenolic compounds, showing potential to be valorized., This study was carried out in the frame of the clabel+ project: Innovative Natural, Nutritious and Consumer Oriented “Clean Label” Foods with the reference POCI-01-0247-FEDER-046080 financed by the Competitiveness and Internationalization Thematic Operational Programme (POCI), under the COMPETE2020, PORTUGAL2020 Partnership Agreement, through the co-financing of European Regional Development Fund (FEDER). The work was supported by UIDB/00211/2020 with funding from FCT/MCTES through national funds. J. D.T. would like to thank to clabel+ project for his fellowship (28/2021/BI). The authors thank Patricia Vicente for her support in the field work. C. Almeida also acknowledges the financial support by LA/P/0045/2020 (ALiCE), UIDB/00511/2020 and UIDP/00511/2020 (LEPABE), funded by national funds through FCT/MCTES (PIDDAC)., info:eu-repo/semantics/publishedVersion

Food waste is commonly defined as a decrease, at all stages of the food system from production to consumption, in mass and/or quality, of food that was originally intended for human consumption, regardless of the cause [1]. To avoid food waste, we aimed to characterize the antioxidant capacity of regional cultivars of apples and pears, for which there are still scarce data. Moreover, different parts of the fruits (peels, seeds and mesocarp) were analysed individually in order to evaluate the potential of by-products to be used a valuable source of natural antioxidants [2], which are believed to be responsible for human health benefits such as anticancer and cardioprotective effects [3]. The objective of this work was to determine the antioxidant properties [4] and fructose content of both by-products (peels and seeds) and edible part of five different Portuguese apple cultivars (Pardo Lindo, Pêro de Borbela, Noiva, Pêro Coimbra and Repinau) and five pear cultivars (Bela-Feia, Torres Novas, Carapinheira Roxa, Lambe-os-Dedos and Amorim), produced in Alcobaça region (Alcobaça), in two consecutive years. The antioxidant properties were evaluated through antioxidant capacity tests (DPPH radical scavenging and -carotene bleaching) and total phenolic content and total flavonoids assays. By-products showed better antioxidant capacity, than the edible part in all cultivars, but the fructose content was very similar in the three parts of the fruits. The results indicate that regional apples´ by-products presented greater potential than regional pears´ by-products. In this line they have potential to be used as a valuable source of antioxidant compounds, which can be further used, by food, cosmetics, food packaging or food supplements industries., This study was carried out in the frame of the clabel+ project: Innovative Natural, Nutritious and Consumer Oriented “Clean Label” Foods with the reference POCI-01-0247-FEDER-046080 financed by the Competitiveness and Internationalization Thematic Operational Programme (POCI), under the COMPETE2020, PORTUGAL2020 Partnership Agreement, through the co-financing of European Regional Development Fund (FEDER). J. D. Teixeira would like to thank to clabel+ project for his fellowship (28/2021/BI). The authors thank Patricia Vicente for her support in the field work. The work was supported by UIDB/00211/2020 with funding from FCT/MCTES through national funds. C. Almeida also acknowledges the financial support by LA/P/0045/2020 (ALiCE), UIDB/00511/2020 and UIDP/00511/2020 (LEPABE), funded by national funds through FCT/MCTES (PIDDAC)., info:eu-repo/semantics/publishedVersion

The authors regret to inform that there are some inaccuracies in the above referred article. More specifically, in the end of the Results and Discussion section 3.6. Anti-Leishmania activity against axenic promastigote and intramacrophagic L. infantum amastigotes it is described the following “Dex-AmB nanocomplexes had an activity similar to the free-drug, despite the slightly higher IC50 values - 0.017 ± 0.009 and 0.023 ± 0.006 μM for Dex-AmB FD and Dex-AmB SD, respectively - promoting a decrease in the amastigote parasite burden in the used concentration range. These results are consistent with the literature where reduced and unreduced AmB-AG conjugates were able to decrease the percentage of macrophages infected with L. infantum (ED50 of 0.035 mg/mL and 0.027 mg/mL, respectively) [75]”. In this sentence the units “mg/mL” are incorrect and the correct ones are “μg/mL”. Moreover, the reference numbered with 75 should be “J. Golenser, et al., Efficacious treatment of experimental leishmaniasis with amphotericin B-arabinogalactan water-soluble derivatives. Antimicrob Agents Chemother 43(9) (1999) 2209-14” instead of “R. Falk, et al. A novel injectable water-soluble amphotericin B-arabinogalactan conjugate. Antimicrob. Agents Chemother., 43 (8) (1999), pp. 1975-1981”. The authors would like to apologise for any inconvenience caused., (undefined), info:eu-repo/semantics/publishedVersion