Isolation of an acetyl-CoA synthetase gene (ZbACS2) from Zygosaccharomyces bailii

A gene homologous to Saccharomyces cerevisiae ACS genes, coding for acetyl-CoA synthetase, has been cloned from the yeast Zygosaccharomyces bailii ISA 1307, by using reverse genetic approaches. A probe obtained by PCR amplification from Z. bailii DNA, using primers derived from two conserved regions of yeast ACS proteins, RIGAIHSVVF (ScAcs1p; 210-219) and RVDDVVNVSG (ScAcs1p; 574-583), was used for screening a Z. bailii genomic library. Nine clones with partially overlapping inserts were isolated. The sequenced DNA fragment contains a complete ORF of 2027 bp (ZbACS2) and the deduced polypeptide shares significant homologies with the products of ACS2 genes from S. cerevisiae and Kluyveromyces lactis (81% and 82% identity and 84% and 89% similarity, respectively). Phylogenetic analysis shows that the sequence of Zbacs2 is more closely related to the sequences from Acs2 than to those from Acs1 proteins. Moreover, this analysis revealed that the gene duplication producing Acs1 and Acs2 proteins has occurred in the common ancestor of S. cerevisiae, K. lactis, Candida albicans, C. glabrata and Debaryomyces hansenii lineages. Additionally, the cloned gene allowed growth of S. cerevisiae Scacs2 null mutant, in medium containing glucose as the only carbon and energy source, indicating that it encodes a functional acetyl-CoA synthetase. Also, S. cerevisiae cells expressing ZbACS2 have a shorter lag time, in medium containing glucose (2%, w/v) plus acetic acid (0.1-0.35%, v/v). No differences in cell response to acetic acid stress were detected both by specific growth and death rates. The mode of regulation of ZbACS2 appears to be different from ScACS2 and KlACS2, being subject to repression by a glucose pulse in acetic acid-grown cells.     

Quantification of ethyl carbamate in cachaça produced in different agro‐industrial production systems

Popularly known by various names such as caninha, dangerous, drips, damned and many other denominations, cachaça is the genuine Brazilian drink, produced by the fermentation of sugarcane juice by yeast, followed by distillation in alambics. Considering that cachaça is the most widely consumed distilled beverage from Brazil, the knowledge of the chemical composition and the presence of potentially toxic compounds such as ethyl carbamate, considered a human carcinogen, is important. The aim of this study was to evaluate the influence of different conditions of the agro‐industrial cachaça production systems, including the variety of sugarcane, on the quantification of ethyl carbamate through the chromatographic. Thirteen unaged beverage samples, produced from different varieties of sugarcane, were analysed. Using analysis of variance and comparison of average concentrations of ethyl carbamate (Scott–Knott, α = 5%), all of the samples were found to contain contaminant levels below the ceiling established by the legislation, which is 210 µg L^?1. Copyright © 2016 The Institute of Brewing & Distilling     

Sacha inchi (Plukenetia volubilis L.) shell: an alternative source of phenolic compounds and antioxidants

Sacha inchi seed (SI) is known as a rich source of oil with high content of polyunsaturared fatty acids of the ω‐3 and ω‐6 type (~85% of total fatty acids). However, few studies have focused on the use of by‐products from the seed. The aim of this study was to characterise the main phenolic families present in SI shell and to evaluate the best extraction solvent for the extraction of phenolic compounds (PC) and antioxidant capacity (AOXC). The PC content corresponded to 74.5 ± 5.1 mg g^?1 of which 93.1% were condensed tannins and the remaining compounds corresponded to free and bound phenolic acids, hydrolyzable tannins, flavonoids and flavanoids. Protocatechic and p‐coumaric acids but also hydroxycinammic acid derivatives of ferulic and o‐coumaric type and lignan derivatives were identified. Acetone containing solvents favoured the extraction of higher amounts of total PC and AOXC. This study highlights the potential use of SI shell as a novel and alternative source of PC antioxidants for the nutraceutical and/or functional food industries.     

Ultrasonic assisted enzymatic digestion (USAED) coupled with high performance liquid chromatography and electrothermal atomic absorption spectrometry as a powerful tool for total selenium and selenium species control in Se-enriched food supplements

The suitability of hyphenated USAED with HPLC separation and ET-AAS determination as a new rapid methodology for Se control in Se-enriched food supplements is demonstrated. Total Se determination and Se speciation are accomplished in a single sample treatment using low sample amounts (ca. 10 mg), and low extracting volume (1 mL). The total Se content in seven of the 10 Se-enriched supplements studied was in agreement with the values obtained after microwave pressurized acid digestion, MW, (test t, p = 0.05). The Se species studied were Se(IV), Se(VI), SeMet, SeMeSeCys, and SeCys₂, being some of the most common found in the 10 supplements studied. Although SeMet was the Se species expected to be present at the highest concentration in most Se-enriched food supplements, we detected it in only three of the 10 samples studied. In the other seven samples, two of them had Se(IV) as the main Se species. The other five supplements had Se species that did not match with any of the five standards selected by us. We have also systematically demonstrated that ultrasonication does not alter the following Se species: Se(IV), Se(VI), SeMet, SeMeSeCys, and SeCys₂. The new procedure can be easily adapted to more Se species and can be routinely used for Se control in Se-enriched food supplements. Concerning the supplements studied, our results suggest that stricter control on the Se content in enriched food supplements in terms of Se species will need to become mandatory.     

Vacuum impregnation of apple slices with Yacon (Smallanthus sonchifolius Poepp. & Endl) fructooligosaccharides to enhance the functional properties of the fruit snack

This work aimed to incorporate prebiotic FOS from yacon in apple slices using vacuum impregnation (VI). Three FOS concentrations (10.3, 14.1 and 18.9 g per 100 g of dry matter (DM)), two temperatures (25 and 35 °C), reuse of extracts and stability of the impregnated slices were evaluated. The highest impregnation level (30.5 g per 100 g DM) was obtained at 35 °C with 14.1% FOS extract while levels of common sugars were reduced. Total phenolics and ABTS antioxidant capacity (AC) slightly decreased while ORAC AC was reduced by 55%. Reuse of the impregnation solution in successive cycles after restoring the FOS level maintained the FOS concentration and profile (GF2–GF7), sugars and phenolic antioxidants. FOS in apple slices remained stable during 4 week storage, while a_w, colour and fracture point changed during storage. This work demonstrated the feasibility of yacon FOS to improve the functional properties of dehydrated apple slices.     

Effect of light,food additives and heat on the stability of sorghum 3-deoxyanthocyanins in model beverages

This work aimed to evaluate the stability of sorghum 3-deoxyanthocyanins (DXA) in model beverages (pH 3.5) elaborated with crude sorghum phenolic extract, containing ascorbic acid and sulphite, under fluorescent light exposure and subjected to heat treatment. There was no significant difference in the DXA degradation during storage under light exposure (24.16%) and absence of light (20.72%). DXA degradation did not differ in the presence of ascorbic acid during storage under light exposure (23.99–25.38%) and absence of light (19.87–21.74%). The addition of sulphite caused an initial bleaching reaction, but as a reversible reaction, the anthocyanin content was higher on the last day of storage compared to the first day. There were no significant differences in total anthocyanin content of all treatments subjected to the heat treatment (80 °C for 5 and 25 min). Thus, crude DXA are very stable under light, additives and heat, and may be useful as natural food colourants.     

Physicochemical and bioactive compounds at edible ripeness of eleven varieties of avocado (Persea americana) cultivated in the Andean Region of Peru

This work aimed evaluating some physicochemical compounds, hydrophilic (H-AC) and lipophilic (L-AC) antioxidant capacity of eleven avocado genotypes grown in the Andes. The results show great variability in physical and chemical properties, dry matter (DM) and lipid contents with ranges of 18.4–39.3% and 10.1–23.2%, respectively. All genotypes had high oleic acid content (50.4–69.9%) and a high unsaturated fatty acid/saturated fatty acid ratio (UFA/SFA, from 3.05 to 6.05), while the unsaponifiable fraction contained mainly β-sitosterol (140.83–235.51 mg/100 g DM) and α-tocopherol (17.44–71.29 µg/g DM). Total phenolic compounds (TPC), H-AC and L-AC ranged from 0.48 to 0.88 mg EAG/g DM, 5.37 to 14.00 and 1.87 to 6.71 μmol ET/g DM, respectively. Although the genotype influences avocado characteristics, climatic conditions in the Andean region seem to be important for the biosynthesis of metabolites of interest and should be considered in future studies.     

The Brazilian Amazonian rainforest harbors a high diversity of yeasts associated with rotting wood,including many candidates for new yeast species

This study investigated the diversity of yeast species associated with rotting wood in Brazilian Amazonian rainforests. A total of 569 yeast strains were isolated from rotting wood samples collected in three Amazonian areas (Universidade Federal do Amazonas-Universidade Federal do Amazonas [UFAM], Piquiá, and Carú) in the municipality of Itacoatiara, Amazon state. The samples were cultured in yeast nitrogen base (YNB)-d -xylose, YNB-xylan, and sugarcane bagasse and corncob hemicellulosic hydrolysates (undiluted and diluted 1:2 and 1:5). Sugiyamaella was the most prevalent genus identified in this work, followed by Kazachstania. The most frequently isolated yeast species were Schwanniomyces polymorphus, Scheffersomyces amazonensis, and Wickerhamomyces sp., respectively. The alpha diversity analyses showed that the dryland forest of UFAM was the most diverse area, while the floodplain forest of Carú was the least. Additionally, the difference in diversity between UFAM and Carú was the highest among the comparisons. Thirty candidates for new yeast species were obtained, representing 36% of the species identified and totaling 101 isolates. Among them were species belonging to the clades Spathaspora, Scheffersomyces, and Sugiyamaella, which are recognized as genera with natural xylose-fermenting yeasts that are often studied for biotechnological and ecological purposes. The results of this work showed that rotting wood collected from the Amazonian rainforest is a tremendous source of diverse yeasts, including candidates for new species.