Exploitation of olive oil mil wastewater for esterase production

[Excerpt] Development of bio-sustainable and renewable resource technologies is extremely important on environmental contexts. Waste can contain many valuable substances and, through a suitable process or technology, this material can be converted into value-added products or raw materials that can be used in secondary processes. To reduce the environmental problems caused by olive oil mill wastewater (OOMW), which is the liquid effluent produced during the extraction process of olive oil, this waste can be applied to bioproducts production. (...

Integrated bioprocess approach for the production of xylooligosaccharides

The demand of prebiotic ingredients has been growing over the years as consumers pay more attention to their health. Xylooligosaccharides (XOS) are considered emergent and competitively priced prebiotics, presenting high potential as food ingredients. As a result, the industry is focused on developing new approaches to improve their production efficiency to meet the increasing demand while reducing costs. Hence, the main purpose of this work was to develop an integrated bioprocess, based on one-step fermentation, for the production of prebiotic XOS, towards the simplification and cost reduction of the process. The one-step fermentation of 13 agro-residues was done using two Trichoderma species. The most promising results were found for T. reesei using brewers spent grain (BSG) as substrate. BSG is an inexpensive and abundant agroindustrial residue that was proven interesting for the production of arabino-xylooligosaccharides (AXOS). In order to reduce the production time obtained with T. reesei (3 d), the Bacillus subtilis 3610 wild type (wt) was successfully used to produce AXOS through direct fermentation of BSG, reducing the production time to 12 h. Genetic engineering was used to further optimize the microorganism performance, by cloning the T. reesei xylanase gene coupled with a secretion tag into the B. subtilis chromosome (B. subtilis 3610 clone 2). This strategy led to a yield increase of 33 % comparing to the wt, and 29 % comparing to the T. reesei. B. subtilis 3610 clone 2 was also selected for downscale production of XOS by direct fermentation of commercial beechwood xylan. The maximum production yield, 306 ± 4 mg/g (XOS/xylan), was achieved after 8 h of fermentation operating under one-time impulse fed-batch regimen. In vitro studies using human fecal inocula were performed to evaluate and compare the potential prebiotic effect of commercial lactulose and the XOS herein produced. The significant increase in the production of short chain fatty acids and CO2, added to the reduction of pH and ammonia concentration suggest that the XOS hold potential functional properties for human health. The results gathered provide important insights for the development of new integrated strategies for XOS production from agro-residues.info:eu-repo/semantics/publishedVersio

Automated image analysis of lung branching morphogenesis from microscopic images of fetal rat explants

Article ID 820214Background. Regulating mechanisms of branching morphogenesis of fetal lung rat explants have been an essential tool for molecular research. This work presents a new methodology to accurately quantify the epithelial, outer contour, and peripheral airway buds of lung explants during cellular development from microscopic images. Methods. The outer contour was defined using an adaptive and multiscale threshold algorithm whose level was automatically calculated based on an entropy maximization criterion. The inner lung epithelium was defined by a clustering procedure that groups small image regions according to the minimum description length principle and local statistical properties. Finally, the number of peripheral buds was counted as the skeleton branched ends from a skeletonized image of the lung inner epithelia. Results. The time for lung branching morphometric analysis was reduced in 98% in contrast to the manual method. Best results were obtained in the first two days of cellular development, with lesser standard deviations. Nonsignificant differences were found between the automatic and manual results in all culture days. Conclusions. The proposed method introduces a series of advantages related to its intuitive use and accuracy, making the technique suitable to images with different lighting characteristics and allowing a reliable comparison between different researchers.The authors acknowledge Foundation for Science and Technology (FCT), Portugal, for the fellowship with the references: SFRH/BD/74276/2010 and SFRH/BPD/46851/2008

Construction of a novel and safe S. cerevisiae biocatalyst for lactulose production

Prebiotics are defined as substrates that are utilized by host microorganisms conferring a health benefit. These compounds have been incorporated in a wide variety of food products1. One of the most well-recognized prebiotics is lactulose. Lactulose is not found naturally so it has to be produced through different methods: chemical or enzymatic synthesis and electro-activation2. Recently, the production of lactulose through lactose isomerization catalyzed by cellobiose 2- epimerase from Caldicellulosiruptor saccharolyticus (CsCE) was reported3. This strategy is gaining attention as a preferable methodology for industrial application due to its notable yields. Saccharomyces cerevisiae, one of the most well-characterized microorganisms, is widely used for the heterologous production of several enzymes, also due to the diverse genetic manipulating tools that are currently available. Here, we propose a new and promising S. cerevisiae biocatalyst. Taking advantage of its GRAS status and using lactose as a single substrate, we believe that it can be a more economic and attractive approach for the synthesis of lactulose. The CsCE gene was cloned in the CEN.PK2-1C S. cerevisiae strain, under TEF or GAP promoters control. Both biocatalysts were used in lactulose production, reaching a concentration of 1.26 g/L (TEF promoter) and 2.15 g/L (GAP promoter). The maximum prebiotic yield was 6.20%. These promising results represent the first use of a S. cerevisiae biocatalyst for lactulose production and demonstrated its potential as a sustainable and safe approach for food application.BBC acknowledge her doctoral grant (SFRH/BD/132324/2017) from the Portuguese Foundation of Science and Technology (FCT). This study was supported by FCT under the scope of the strategic funding of UID/BIO/04469/2019 unit, the Project FoSynBio (POCI-01-0145-FEDER-029549) and BioTecNorte operation (NORTE-01-0145-FEDER000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte.info:eu-repo/semantics/publishedVersio

Screening of fungal sources of -galactosidase with potential for the synthesis of prebiotics

Book of Abstracts of CEB Annual Meeting 2017[Excerpt] β-Galactosidases (EC 3.2.1.23), also known as lactases, are a family of enzymes able to catalyse two different types of reactions, namely hydrolysis and transgalactosylation. The hydrolytic activity is commonly applied in the food industries to reduce the lactose content of dairy products, preventing lactose crystallization problems and increasing sweetness, flavour and solubility. On the other hand, transgalactosylation reactions have been explored in the synthesis of lactose-based prebiotics, such as galacto-oligosaccharides (GOS), lactosucrose [1] and lactulose [2], with potential application in the pharmaceutical and food industry. These prebiotics are enzymatically produced through the hydrolysis of lactose and further transfer of a galactosyl residue to a suitable acceptor, i.e. fructose for the disaccharide lactulose; sucrose for the trisaccharide lactosucrose; and lactose for GOS. The sources of βgalactosidase are extensively distributed in nature, namely in microorganisms, plants and animal organs. [...]info:eu-repo/semantics/publishedVersio

Enzymatic synthesis of lactulose using a novel biocatalyst

info:eu-repo/semantics/publishedVersio

Study of the geomorphological and archaeological aspects of Sintra area (Portugal) as contribution to its tourist appraisal and promotion

The Portughese town of Sintra is an UNESCO world heritage site for itscultural and environmental aspects. The town is located 30 km from Lisbon and 15km from the Atlantic Ocean, and is a touristic site in every seasons mainly for itsarchitectural qualities. This work describes the geomorphological and archaeologicalaspects of the town and the surrounding Serra de Sintra in order to give an addedvalue to tourist appraisal and promotion of the area. Serra de Sintra is an ellipticigneous massif 10 km E-W and 5 km N-S, 300-500 m above sea level. The geologicalstructure is complex but it can be simplified as a core of sienites surrounded bygranites intruded in a limestone plateau. The morphogenetic processes, besidecommon landforms such as narrow valleys, scarps and ridges, shaped a variety ofparticular granite morphologies such as round block fields, inselbergs, castle koppies,etc…. The most spectacular ones have been described and classified as geosites in adata-base. The oldest archaeological remains dates to the Mesolithic, but the area hasbeen extensively occupied during the Neolitic/Calcolithic, Bronze and Iron Ages. Inhistorical times Romans, Visigots and Arabs left some interesting architectural andlinguistic marks. All the archaeological sites have been classified according tobibliographic, museal data and field surveys. They are also all included in a data-base.GeoArchaeo-Tourist maps at 1:25,000 and 1.10,000 scales of the Serra de Sintra andSintra town are the information results collected in the geomorphological andcultural data-bases. The maps have been implemented, by means of an ArcGIScomputer programme, integrating geological, geomorphological, archaeological andhistorical aspects with the tourist infrastructures (information points, parking areas,accommodations, panoramic points, etc.) in order to obtain documents readable, simple, clear but scientifically accurate also for non-expert users. This study could besuitably simplified and summarized in a guide book, with enclosed GeoArchaeo-Tourist maps, as a contribution to improve the knowledge and appraisal of the Sintraterritory also for its landscape and archaeological aspects

Monitoring nitrogen nutritional status of vegetables in the social garden of Polytechnic Institute of Bragança, NE Portugal

Urban agriculture is a global phenomenon. In developing countries urban agriculture has had crucial importance in alleviating problems of extreme poverty and unemployment of the populations of the larger cities. In developed countries has been particularly important during periods of economic depression. Nowadays, the urban agriculture in developed countries is increasing, aimed more at leisure, sports and recreation of the urban dwellers. These spaces are also emphasizing environmental education awareness by implementing environmentally friendly agricultural practices. In the case of Portugal, urban agriculture is frequently regulated by norms, implicitly or explicitly, similar to those established for organic farming. The urban agriculture project of the Polytechnic Institute of Bragança (IPB) is one of these kinds of projects in which participants are encouraged to use sustainable farming practices. The fertilizers used are usually farmyard manures and other organic wastes. This work takes part of a larger project that aims to monitor the nutritional status of the plants, the residual mineral nitrogen content in the soil and the contamination of plants and soils with heavy metals. In this report, it will be present results only for the first two goals

Biocatalytic approaches using lactulose: end product compared with substrate

Lactulose is a lactose-based carbohydrate with well-known prebiotic effect and recognized medical applications. Currently, the commercially available lactulose is chemically synthesized. Nevertheless, the process leads to low yields and high levels of by-products. Alternatively, lactulose can be produced by enzymatic synthesis, which provides a cleaner production under mild conditions. Two different enzymatic routes were reported for lactulose production. Lactulose can be obtained through hydrolysis and transfer reactions catalyzed by a glycosidase. Alternatively, lactulose can be produced by direct isomerization of lactose to lactulose catalyzed by cellobiose-2-epimerase. An interesting characteristic of lactulose is also its capacity to act as substrate in additional enzymatic synthesis which leads to the formation of attractive compounds, such as lactulose-based oligosaccharides and lactulose esters. Besides increasing the interest and potential of lactulose, these lactulose-based compounds can also offer new and promising functionalities and applications. Herein, we review the enzymes involved in the synthesis of lactulose, as well as the reaction conditions and yields. The potential of different enzymes is discussed and it is shown that reaction conditions and composition of products depend on the type of enzyme and its microbial source. The conversion of lactulose into lactulose-based compounds is also covered, describing in detail the biocatalysts involved, the reaction conditions used, and the potential of the final products obtained.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684). The authors also thank the FCT for the financial support under the scope of the Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462). SCS also acknowledges her post doc grant (SFRH/BPD/88584/2012) from FCT

Perspectives on the biotechnological production and potential applications of lactosucrose: A review

Lactosucrose is a synthetic trisaccharide composed of galactose, glucose and fructose. This compound is obtained through enzymatic synthesis using lactose and sucrose as substrates. The enzymes involved in the process are able to catalyse both hydrolysis and transfer reactions. The yield and productivity of the process are usually affected by the occurrence of parallel hydrolysis of the newly formed product (lactosucrose). Therefore, it is important to find efficient strategies to avoid or minimize product degradation. Furthermore, in the last decades the demand for lactosucrose has significantly increased. This compound is considered a potential prebiotic and several beneficial effects associated to its consumption have been described. As a result, it has been included in the formulations of functional foods. This review covers the most relevant information about lactosucrose, including its synthesis and purification, beneficial effects at physiological level, and also its potential applications.Fundação para a Ciência e a Tecnologia (FCT) Strategic Project of UID/BIO/04469/2013 unit, the project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462) and the project “BioInd – Biotechnology and Bioengineering for im- proved Industrial and Agro-Food processes”, REF. NORTE-07- 0124-FEDER-000028 co-funded by the Programa Operacional Regional do Norte (ON. 2–ONovo Norte), QREN, FEDER