ATLANTIC-PRIMATES: a dataset of communities and occurrences of primates in the Atlantic Forests of South America

Primates play an important role in ecosystem functioning and offer critical insights into human evolution, biology, behavior, and emerging infectious diseases. There are 26 primate species in the Atlantic Forests of South America, 19 of them endemic. We compiled a dataset of 5,472 georeferenced locations of 26 native and 1 introduced primate species, as hybrids in the genera Callithrix and Alouatta. The dataset includes 700 primate communities, 8,121 single species occurrences and 714 estimates of primate population sizes, covering most natural forest types of the tropical and subtropical Atlantic Forest of Brazil, Paraguay and Argentina and some other biomes. On average, primate communities of the Atlantic Forest harbor 2 ± 1 species (range = 1–6). However, about 40% of primate communities contain only one species. Alouatta guariba (N = 2,188 records) and Sapajus nigritus (N = 1,127) were the species with the most records. Callicebus barbarabrownae (N = 35), Leontopithecus caissara (N = 38), and Sapajus libidinosus (N = 41) were the species with the least records. Recorded primate densities varied from 0.004 individuals/km 2 (Alouatta guariba at Fragmento do Bugre, Paraná, Brazil) to 400 individuals/km 2 (Alouatta caraya in Santiago, Rio Grande do Sul, Brazil). Our dataset reflects disparity between the numerous primate census conducted in the Atlantic Forest, in contrast to the scarcity of estimates of population sizes and densities. With these data, researchers can develop different macroecological and regional level studies, focusing on communities, populations, species co-occurrence and distribution patterns. Moreover, the data can also be used to assess the consequences of fragmentation, defaunation, and disease outbreaks on different ecological processes, such as trophic cascades, species invasion or extinction, and community dynamics. There are no copyright restrictions. Please cite this Data Paper when the data are used in publications. We also request that researchers and teachers inform us of how they are using the data. © 2018 by the The Authors. Ecology © 2018 The Ecological Society of Americ

Purification and characterization of α-galactosidase from Tachigali multijuga seeds and partial cloning of soybean stachyose synthase gene

A soja, devido ao seu alto valor nutricional, é mundialmente consumida na forma de óleo, margarinas, proteína texturizada e extrato hidrossolúvel. O extrato hidrossolúvel, conhecido como leite de soja, é uma alternativa alimentar para muitas pessoas com intolerância à lactose. No entanto, a presença dos galactooligossacarídeos (GO) restringe o consumo de soja e seus produtos derivados. A mucosa intestinal do homem e animais monogástricos não possui a enzima α-galactosidase necessária para hidrolisar as ligações α-1,6 dos resíduos de galactose presentes nos GO, desta forma estes oligossacarídeos não são digeridos causando diarréia, náuseas, e flatulência. Como parte de nosso esforço para aumentar o consumo de soja e produtos derivados pelo homem, este trabalho teve dois objetivos: purificar e caracterizar uma α-galactosidase de sementes de Tachigali multijuga e avaliar sua capacidade de hidrolisar os GO em leite de soja; e isolar, clonar e seqüênciar parte do gene que codifica a enzima estaquiose sintase (STS) de soja, para ser usada para seu silenciamento por técnicas de engenharia genética. A enzima α-galactosidase foi purificada por precipitação ácida, diálise, cromatografias de troca iônica e filtração em gel. A fração enzimática purificada foi analisada por eletroforese em gel SDS-PAGE e revelou uma única banda protéica com massa molecular de 38 kDa. Atividade máxima da α-galactosidase foi detectada em pH 5,0 - 5,5 a 50 °C. A enzima foi estável no pH 4,5 - 7,0 por 15 min a 40 ºC, mas perdeu total atividade no pH 7,0 por 30 min a 40 ºC. Cerca de 65 % de atividade original foi mantida após préincubação por 200 h a 35 ºC. A meia-vida da α-galactosidase a 40 ºC foi 17,6 h. A α-galactosidase apresentou especificidade absoluta para galactose ligada em posição α, e ela foi completamente inibida por SDS, Hg2+, Cu2+ e Ag+, e parcialmente inibida por D-galactose e melibiose. Os valores de KM para hidrólise do ρNPαGal, melibiose, rafinose e estaquiose foram 0,45; 5,37; 39,62 e 48,80 mM, respectivamente. A α-galactosidase foi inibida competitivamente por galactose quando o ρNPαGal foi usado como substrato, com Ki de 2,74 mM. Os valores de energia de ativação estimados para os substratos ρNPαGal e rafinose foram 13,86 e 4,75 kcal/mol, respectivamente. O tratamento da farinha desengordurada de soja com a α-galactosidase purificada de sementes de Tachigali multijuga resultou em 72 e 50 % de redução do conteúdo de rafinose e estaquiose, respectivamente, após incubação por 4 h a 40 °. A enzima apresentou capacidade para hidrolisar goma de alfarroba e goma guar, sugerindo possível aplicação industrial para promover propriedades gélicas em polissacarídeos. A atividade de STS foi determinada durante o desenvolvimento da semente de soja e em sementes maduras. A quantificação de estaquiose foi feita por HPLC. O conteúdo de estaquiose na soja madura foi 4,10 % e a atividade específica da enzima STS foi 2,15 nkat/mg, usando rafinose e galactinol como substratos. Um fragmento do gene STS foi isolado por PCR usando cDNA de semente em combinação com primers degenerados. A análise do padrão de expressão por RT-PCR mostrou que o gene STS é expresso em todos os estádios de desenvolvimento do grão, além de folhas, caule e raiz. A clonagem de um fragmento de 983 pb no vetor pGEM-T Easy foi confirmada por seqüenciamento. A identidade do fragmento, usando a ferramenta BLAST, confirmou que a seqüência clonada se refere ao gene STS, que até então não tinha sido isolada e seqüenciada em soja. Este trabalho abre perspectivas para pesquisas biotecnológicas visando o desenvolvimento de variedades de soja, com reduzido conteúdo de estaquiose e então mais adequadas para o consumo humano.Because of its high nutritional value, soybean is worldwide consumed in the forms of oil, margarines, texturized protein and hydrosoluble extract. The hydrosoluble extract, known as soymilk, is also an alternative to dairy food for many lactose-intolerant people. However, the presence of galactooligosaccharides (GO) restricts the consumption of soybean and soyderived products. The intestinal mucous membrane of humans and monogastric animals lacks the α-galactosidase enzyme that is necessary for the hydrolysis of α-1,6 bonds of galactosil residues present in GO, consequently these oligosaccharides are not digested causing diarrhea, nauseas and flatulence. As part of our effort to increase the consumption of soybean and soy-derived products by humans, this work was developed with two goals: to purify and characterize one α-galactosidase from Tachigali multijuga seeds and evaluate its capacity for hydrolysing GO in soymilk; and to isolate, clone and sequence part of the gene that encodes the soybean stachyose synthase enzyme (STS), to be used for its silencing by genetic engineering techniques. The α-galactosidase enzyme was purified by acid precipitation, dialysis, ion exchange and gel filtration chromatographies. The purified enzymatic fraction was analyzed by electrophoresis in SDS-PAGE gel revealing only one protein band with estimated molecular mass of 38 kDa. Maximal α-galactosidase activity was detected at pH 5.0 5.5 and 50 ºC. The enzyme was stable at pH 4.5 - 7.0 at 40 ºC for 15 min, but lost its activity after incubation for 30 min at pH 7.0 at 40 ºC. About 65 % of original activity was kept after pre-incubation at 35 ºC for 200 h. Half-life of the α-galactosidase at 40 ºC was 17.6 h. The α-galactosidase showed absolute specificity for galactose linked to the α position, and it was completely inhibited by SDS, Hg2+, Cu2+ and Ag+ and partially inhibited by D-galactose and melibiose. The KM values for the hydrolysis of ρNPαGal, melibiose, raffinose and stachyose were 0.45, 5.37, 39.62 and 48.80 mM, respectively. The enzyme was inhibited competitively by galactose when ρNPαGal was used as substrate, with Ki of 2.74 mM. Activation energies estimated for ρNPαGal and raffinose were 13.86 and 4.75 kcal/mol, respectively. The treatment of defatted soybean flour with the α-galactosidase purified from Tachigali multijuga seeds resulted in 72 and 50 % of reduction in raffinose and stachyose contents, respectively, after incubation at 40 ºC for 4 h. The enzyme showed capacity to hydrolyze the locust bean gum and guar gum, suggesting that it can be used in the industry to improve gelling properties in polysaccharides. STS activity was determined during the development of soybean seed and in mature seeds. Stachyose quantification was carried out by HPLC. Stachyose content in mature soybean seeds was 4.10% and STS specific activity was 2.15 nkat/mg, using raffinose and galactinol as substrates. A fragment of the STS gene was isolated by PCR using seed cDNA in combination with degenerated primers. The analysis of the expression patterns by RT-PCR showed that the STS gene is expressed in all seed developmental stages, as well as in leaves, stem and roots. The cloning of a 983 base-pair fragment in a pGEM-T Easy vector was confirmed by sequencing. The identity of the fragment, using BLAST analysis, confirmed that the cloned sequence referred to the STS gene, which had not been isolated and sequenced in soybeans until then. This work opens perspectives for biotechnological researches aiming at the development of soybean varieties with reduced stachyose content and therefore more suitable for human consumption.Coordenação de Aperfeiçoamento de Pessoal de Nível Superio

Purification and characterization of Aspergillus terreus α-Galactosidases and their use for hydrolysis of Soymilk Oligosaccharides

α-Galactosidases has the potential to hydrolyze α-1-6 linkages in raffinose family oligosaccharides (RFO). Aspergillus terreus cells cultivated on wheat bran produced three extracellular forms of α-galactosidases (E1, E2, and E3). E1 and E2 α-galactosidases presented maximal activities at pH 5, while E3 α-galactosidase was more active at pH 5.5. The E1 and E2 enzymes showed stability for 6 h at pH 4–7. Maximal activities were determined at 60, 55, and 50°C, for E1, E2, and E3 α-galactosidase, respectively. E2 α-galactosidase retained 90% of its initial activity after 70 h at 50°C. The enzymes hydrolyzed ρNPGal, melibiose, raffinose and stachyose, and E1 and E2 enzymes were able to hydrolyze guar gum and locust bean gum substrates. E1 and E3 α-galactosidases were completely inhibited by Hg2+, Ag+, and Cu2+. The treatment of RFO present in soy milk with the enzymes showed that E1 α-galactosidase reduced the stachyose content to zero after 12 h of reaction, while E2 promoted total hydrolysis of raffinose. The complete removal of the oligosaccharides in soy milk could be reached by synergistic action of both enzyme

A Chrysoporthe cubensis enzyme cocktail produced from a low-cost carbon source with high biomass hydrolysis efficiency

Não conta o número de páginas.Low cost and high efficiency cellulolytic cocktails can consolidate lignocellulosic ethanol technologies. Sugarcane bagasse (SCB) is a low cost agro-industrial residue, and its use as a carbon source can reduce the costs of fungi cultivation for enzyme production. Chrysoporthe cubensis grown under solid state fermentation (SSF) with wheat bran has potential to produce efficient enzymatic extracts for SCB saccharification. This fungus was grown under submersed fermentation (SmF) and SSF with in natura SCB, pretreated with acid or alkali and with others carbon sources. In natura SCB induced the highest carboxymethylcellulase (CMCase), xylanase, β-xylosidase, α-galactosidase and mannanase activities by C. cubensis under SSF. In natura and washed SCB, inducers of enzyme production under SSF, did not induce high cellulases and hemicellulases production by C. cubensis in SmF. The C. cubensis enzymatic extract produced under SSF with in natura SCB as a carbon source was more efficient for lignocelulolic biomass hydrolysis than extracts produced under SSF with wheat bran and commercial cellulolytic extract. Chrysoporthe cubensis showed high potential for cellulases and hemicellulases production, especially when grown under SSF with in natura SCB as carbon source

Characterization and biotechnological application of an acid α-galactosidase from Tachigali multijuga Benth. seeds

Tachigali multijuga Benth. seeds were found to contain protein (364 mg g−1 dwt), lipids (24 mg g−1 dwt), ash (35 mg g−1 dwt), and carbohydrates (577 mg g−1 dwt). Sucrose, raffinose, and stachyose concentrations were 8.3, 3.0, and 11.6 mg g−1 dwt, respectively. α-Galactosidase activity increased during seed germination and reached a maximum level at 108 h after seed imbibition. The α-galactosidase purified from germinating seeds had an Mr of 38,000 and maximal activity at pH 5.0–5.5 and 50 °C. The enzyme was stable at 35 °C and 40 °C, but lost 79% of its activity after 30 min at 50 °C. The activation energy (Ea) values for p-nitrophenyl-α-d-galactopyranoside (pNPGal) and raffinose were 13.86 and 4.75 kcal mol−1, respectively. The Km values for pNPGal, melibiose, raffinose, and stachyose were 0.45, 5.37, 39.62 and 48.80 mM, respectively. The enzyme was sensitive to inhibition by HgCl2, SDS, AgNO3, CuSO4, and melibiose. d-Galactose was a competitive inhibitor (Ki = 2.74 mM). In addition to its ability to hydrolyze raffinose and stachyose, the enzyme also hydrolyzed galactomannan

Biochemical composition and indigestible oligosaccharides in Phaseolus vulgaris L. seeds

Common beans have a high nutritional value, but contain galactooligosaccharides (GO), which cause flatulence and intestinal discomfort in humans. The biochemical composition of ten bean cultivars was determined to select those of high protein and low GO contents. The cultivars varied in carbohydrate (47.02–60.17%), GO (3.12 – 5.71%), protein (22.17–33.50%), lipid (1.13–1.81%), moisture (11.42–12.93%) and ash contents (4.08–5.61%). ‘Mexico 222’ presented the highest α-galactosidase activity. Protein and GO contents were positively correlated. ‘Perry Marrow’ combined high protein and low GO concentrations, indicating it can be used in improvement programs aiming at high-quality cultivars for human consumption