The cyclic peptide G4CP2 enables the modulation of galactose metabolism in yeast by interfering with GAL4 transcriptional activity

Genetically-encoded combinatorial peptide libraries are convenient tools to identify peptides to be used as therapeutics, antimicrobials and functional synthetic biology modules. Here, we report the identification and characterization of a cyclic peptide, G4CP2, that interferes with the GAL4 protein, a transcription factor responsible for the activation of galactose catabolism in yeast and widely exploited in molecular biology. G4CP2 was identified by screening CYCLIC, a Yeast Two-Hybrid-based combinatorial library of cyclic peptides developed in our laboratory. G4CP2 interferes with GAL4-mediated activation of galactose metabolic enzymes both when expressed intracellularly, as a recombinant peptide, and when provided exogenously, as a chemically-synthesized cyclic peptide. Our results support the application of G4CP2 in microbial biotechnology and, additionally, demonstrate that CYCLIC can be used as a tool for the rapid identification of peptides, virtually without any limitations with respect to the target protein. The possible biotechnological applications of cyclic peptides are also discussed

HISTONE DEACETYLASE19 Controls Ovule Number Determination and Transmitting Tract Differentiation

The gynoecium is critical for the reproduction of flowering plants as it contains the ovules and the tissues that foster pollen germination, growth, and guidance. These tissues, known as the reproductive tract (ReT), comprise the stigma, style, and transmitting tract (TT). The ReT and ovules originate from the carpel margin meristem (CMM) within the pistil. SHOOT MERISTEMLESS (STM) is a key transcription factor for meristem formation and maintenance. In all above-ground meristems, including the CMM, local STM downregulation is required for organ formation. However, how this downregulation is achieved in the CMM is unknown. Here, we have studied the role of HISTONE DEACETYLASE 19 (HDA19) in Arabidopsis (Arabidopsis thaliana) during ovule and ReT differentiation based on the observation that the hda19-3 mutant displays a reduced ovule number and fails to differentiate the TT properly. Fluorescence-activated cell sorting (FACS) coupled with RNA-sequencing (RNA-seq) revealed that in the CMM of hda19-3 mutants, genes promoting organ development are downregulated while meristematic markers, including STM, are upregulated. HDA19 was essential to downregulate STM in the CMM, thereby allowing ovule formation and TT differentiation. STM is ectopically expressed in hda19-3 at intermediate stages of pistil development, and its downregulation by RNA interference alleviated the hda19-3 phenotype. Chromatin immunoprecipitation assays indicated that STM is a direct target of HDA19 during pistil development and that the transcription factor SEEDSTICK is also required to regulate STM via histone acetylation. Thus, we identified factors required for the downregulation of STM in the CMM, which is necessary for organogenesis and tissue differentiation

Bioeconomy perception by future stakeholders: Hearing from European forestry students

This article provides useful information for universities offering forestry programs and facing the growing demand for bioeconomy education. An explorative survey on bioeconomy perception among 1400 students enrolled in 29 universities across nine European countries offering forestry programs was performed. The data have been elaborated via descriptive statistics and cluster analysis. Around 70% of respondents have heard about the bioeconomy, mainly through university courses. Students perceive forestry as the most important sector for bioeconomy; however, the extent of perceived importance of forestry varies between countries, most significantly across groups of countries along a North–South European axis. Although differences across bachelor and master programs are less pronounced, they shed light on how bioeconomy is addressed by university programs and the level of student satisfaction with this. These differences and particularities are relevant for potential development routes towards comprehensive bioeconomy curricula at European forestry universities with a forestry focus.Peer reviewe

Public perceptions of using forests to fuel the European bioeconomy: Findings from eight university cities

The political project on bioeconomy strives to address multiple societal aspirations, namely combine economic growth with environmental sustainability in some socially acceptable manner. The contradictions between the goals and the concrete plans to increase production, processing, and consumption of forest biomass in Europe have however raised sustainability concerns within and beyond its borders. While political actors articulate such contradictions differently and compete for traction for their viewpoints in the public discourse, little is known about how citizens of urban areas perceive this discourse. Conceptualising perception as a multidimensional construct, data from eight European university cities (Bordeaux, Bratislava, Freiburg, Helsinki, Padua, St. Petersburg, Uppsala, Vienna) are statistically analysed to explore its dimensions, the communities of like-minded citizens forming across those dimensions, and the traits associating with membership in each such community. Five communities across six dimensions from biocentrism through distributional aspects to adherence to political goals are identified: adherent-environmentalist, adherent-governmentalist, critical-reformist, critical-agriculturalist, and indifferent. City of residence and perceived familiarity with bioeconomy clearly interact with perception. There is however considerable variation in communities within and across the eight cities, suggesting deeper social tension beyond the public discourse. Much of the within-community variation remains unexplained, though, calling for more work locally. Implications for forest policy are derived.Peer reviewe

Microusinas de etanol de batata-doce : viabilidade econômica e técnica

Fatores ambientais e econômicos impulsionam mundialmente a produção de biocombustíveis. No Brasil, o etanol, produzido da cana-de-açúcar, já é um biocombustível estabelecido e substitui ca. 40% da gasolina, representando 13% do total de energia necessária para transportes. Nesse cenário, o Rio Grande do Sul (RS) é um grande comprador de etanol, tendo produzido nos últimos anos apenas 2% do consumo estadual de etanol hidratado combustível. O estado também consome em média 600 milhões de litros de etanol anidro por ano, adicionados na proporção de 25% à gasolina comum, e, a partir de 2010, 460 milhões de litros de etanol por ano para a produção de polietileno verde. Essa conjuntura demonstra uma oportunidade para aumentar a produção local de etanol. O presente trabalho buscou primeiramente avaliar a viabilidade da produção de etanol no Rio Grande do Sul em um modelo de microusinas descentralizadas (ca. 1.000 L.dia-1). Para tanto, foram empregados os indicadores econômicos, como valor presente líquido, taxa interna de retorno de investimento e tempo de retorno de investimento. Foram comparados cenários que empregaram apenas cana-de-açúcar e combinações de cana-de-açúcar, sorgo sacarino, mandioca e batata-doce. A utilização de cana-de-açúcar sem o consórcio com outra cultura se mostrou inviável, exceto quando mais de 40% ou 80% da produção, para as produtividades de 80 e 50 t.ha-1 respectivamente, é destinada ao consumo próprio. Entre os cenários com combinação de culturas, aqueles que combinaram sorgo com cana-de-açúcar e sorgo com batata-doce foram os únicos que se mostraram viáveis quando toda a produção foi destina à venda para terceiros. Quando produtividades médias de cana-de-açúcar próximas a 80 t.ha-1 podem ser alcançadas, verificou-se que a combinação dessa cultura com sorgo sacarino apresentou o melhor potencial entre os cenários avaliados. Já para regiões onde esses valores não são atingidos, o consórcio de sorgo sacarino e batata-doce se mostrou a melhor opção. Posteriormente, foram realizados experimentos em shaker para estudar a influência da concentração de substrato e da proporção de enzima na hidrólise a frio da batata-doce, determinar o melhor pré-tratamento, verificar a necessidade de suplementação do meio e do controle de pH na condução das hidrólises e fermentações simultâneas e finalmente testar a melhor condição em biorreator. Para hidrolisar o amido, empregou a mistura de enzimas Stargen™ 002 e, para suplementar o meio, o fertilizante NITROFOS KL. Em todos os experimentos, usou-se a cultivar BRS Cuia, cuja caracterização indicou teor de carboidratos de 28,7%, possibilitando a produção de 185 L.t-1 de etanol e 7.400 L. ha-1. A metodologia de superfície de resposta indicou a condição 200 g.L-1 de batata-doce e 45 GAU.g de batata–doce-1 como a que apresentou o melhor compromisso entre alta taxa de formação de glicose na primeira hora (8,3 g.L-1.h-1) e baixo consumo de enzimas. O pré-tratamento de uma hora que levou a maior concentração de glicose (14,3 g.L-1) foi na temperatura de 52°C na presença da mistura de enzimas. O estudo da hidrólise e fermentação simultâneas mostrou que a suplementação do meio não apresenta influência significativa, enquanto o controle de pH aumentou em aproximadamente 40% a produção de etanol. Os testes em biorreator reproduziram os resultados anteriores mesmo sendo realizados em ambiente semiestéril, que se aproxima da condição industrial.In Brazil, sugarcane ethanol is already a reality as a biofuel and replaces 40% of gasoline, meaning 13% of energy for transportation. In this scenario, Rio Grande do Sul has produced only 2% of the annual demand for hydrate ethanol in the last years; therefore it is a big importer of ethanol from other states. Additionally, it consumes every year 600 million liters of anhydrous ethanol mixed in the gasoline and 460 million liters for production of green plastic. These numbers highlight the opportunity of producing ethanol locally. In regard to these facts, the present work aimed to evaluate the feasibility of producing ethanol in small scale plants (ca. 1 000 L.dia-1) in Rio Grande do Sul. For this purpose, economic indicators, such as net present value, internal rate of return and payback period were employed. The compared scenarios involved combinations of sugar cane, sweet sorghum, cassava and sweet potato. When only sugarcane was used, the scenarios for 80 and 50 t.ha-1 were unfeasible if less than 40% or 80% of the production, respectively, was used by the own producer. Regarding the scenarios with mix of crops, the mix of sweet sorghum with sugar cane and sweet sorghum with sweet potato presented the best performances. For regions, where 80 t.ha-1 of sugar cane productivity can be achieved, it was verified that the first combination leads to the best result. For other regions, the combination of sweet sorghum and sweet potato presented itself as the more feasible scenario. As a consequence, in the second part of this work, the cold hydrolysis and fermentation of sweet potato was experimentally studied. For that, it was employed the sweet potato strain BRS Cuia, whose carbohydrate level reaches 28,7%. What it can be translated into a potential to produce 185 L.t-1 ethanol, or equivalently 7 400 L. ha-1. The enzymes blend adopted for the hydrolysis stage was Stargen™ 002, while the fertilizer NITROFOS KL was used for fermentation medium supplementation. The surface response method indicated 200 g.L-1 of sweet potato and 45 GAU.g of sweet potato-1 as the best balance between high glucose formation rate in the first hour (8,3 g.L-1.h-1) and low enzyme consume. The one hour pre-treatment that achieved the highest glucose concentration (14,3 g.L-1) was at 52°C in the presence of the enzymes blend. The study of the simultaneous hydrolysis and fermentation showed that the medium supplementation has no significant effect over the fermentation performance, while the pH control is beneficial, improving the ethanol production in 40%. Finally, the tests in bioreactor could reproduce the previous results, even though the experiments were carried out non-sterilely