Enzymatic extraction of hydroxycinnamic acids from coffee pulp

Ferulic, caffeic, p-coumaric and chlorogenic acids are classified as hydroxycinnamic acids, presenting anticarcinogenic, anti-inflammatory and antioxidant properties. In this work, enzymatic extraction has been studied in order to extract high value-added products like hydroxycinnamic acids from coffee pulp. A commercial pectinase and enzyme extract produced by Rhizomucor pusillus strain 23aIV in solid-state fermentation using olive oil or coffee pulp (CP) as an inducer of the feruloyl esterase activity were evaluated separately and mixed. The total content (covalently linked and free) of ferulic, caffeic, p-coumaric and chlorogenic acids was 5276 mg per kg of coffee pulp. Distribution was as follows (in %): chlorogenic acid 58.7, caffeic acid 37.6, ferulic acid 2.1 and p-coumaric acid 1.5. Most of the hydroxycinnamic acids were covalently bound to the cell wall (in %): p-coumaric acid 97.2, caffeic acid 94.4, chlorogenic acid 76.9 and ferulic acid 73.4. The content of covalently linked hydroxycinnamic acid was used to calculate the enzyme extraction yield. The maximum carbon dioxide rate for the solid-state fermentation using olive oil as an inducer was higher and it was reached in a short cultivation time. Nevertheless, the feruloyl esterase (FAE) activity (units per mg of protein) obtained in the fermentation using CP as an inducer was 31.8 % higher in comparison with that obtained in the fermentation using olive oil as the inducer. To our knowledge, this is the first report indicating the composition of both esterified and free ferulic, caffeic, p-coumaric and chlorogenic acids in coffee pulp. The highest yield of extraction of hydroxycinnamic acids was obtained by mixing the produced enzyme extract using coffee pulp as an inducer and a commercial pectinase. Extraction yields were as follows (in %): chlorogenic acid 54.4, ferulic acid 19.8, p-coumaric acid 7.2 and caffeic acid 2.3. An important increase in the added value of coffee pulp was mainly due to the extraction of chlorogenic acid

Challenges of accessibility of a community heritage tourist route: The Route of the Caste War

This article presents the results of an accessibility analysis of The Caste War Route (RGC), prior to its commercialization as a community heritage product. The analysis consists of a diagnosis of the resource to establish destination-planning strategies. The accessibility diagnosis goes beyond adapting physical spaces for transit, considering that the resource is accessible to all types of people, including economic, spatial and temporal accessibility, criteria on which the research focuses. The diagnosis was prepared through a multidisciplinary investigation that collected information from different sectors with qualitative and quantitative tools that combined the recording of data and the opinion of the residents of the area, key informants; Government officials, museum workers, tourism service providers, non-governmental organizations and visitors were included in this research. Accessibility is a multivariate concept; its analysis required an instrument with cultural indicators distributed in categories, which provides objective, rigorous and relevant information. The research approach was qualitative, including Participatory Action Research and ethnographic techniques such as participant observation (PAR), interviews and document review as part of the process. It is necessary to propose promotional strategies focused on rural cultural products, that disseminate the sites and activities considered heritage by the community, and that the inhabitants are willing to share with visitors, so that local hosts are the ones who offer this service. The necessary strategies are the equal participation and involvement of women and men, the participation of students and academics in training courses and orientation to local service providers. These products face important challenges: they must differentiate themselves from others to build a unique local identity, and at the same time, form alliances with other local communities to create and strengthen local tourism products and services to create a complete touristic experience versus isolated experiences in individual communities. Achieving integration is essential for a successful project and the regional growth and development of the localities involved

Optimisation of biomass, exopolysaccharide and intracellular polysaccharide production from the mycelium of an identified Ganoderma lucidum strain QRS 5120 using response surface methodology

Wild-cultivated medicinal mushroom Ganoderma lucidum was morphologically identified and sequenced using phylogenetic software. In submerged-liquid fermentation (SLF), biomass, exopolysaccharide (EPS) and intracellular polysaccharide (IPS) production of the identified G. lucidum was optimised based on initial pH, starting glucose concentration and agitation rate parameters using response surface methodology (RSM). Molecularly, the G. lucidum strain QRS 5120 generated 637 base pairs, which was commensurate with related Ganoderma species. In RSM, by applying central composite design (CCD), a polynomial model was fitted to the experimental data and was found to be significant in all parameters investigated. The strongest effect (p lt 0.0001) was observed for initial pH for biomass, EPS and IPS production, while agitation showed a significant value (p lt 0.005) for biomass. By applying the optimized conditions, the model was validated and generated 5.12 g/L of biomass (initial pH 4.01, 32.09 g/L of glucose and 102 rpm), 2.49 g/L EPS (initial pH 4, 24.25 g/L of glucose and 110 rpm) and 1.52 g/L of IPS (and initial pH 4, 40.43 g/L of glucose, 103 rpm) in 500 mL shake flask fermentation. The optimized parameters can be upscaled for efficient biomass, EPS and IPS production using G. lucidum

Fronteras in biotecnologia y bioingenieria

The low water activity, the complex nature of the support and/or substrate, the concentation of substrates used and products formed, and the heterogeneity within the system, influence the growth and physiology of the microbial strains used in solid state fermentation (SSF). As a result, the quality and quantity of the products formed from SSF processes significantly differ from those of submerged fermentation (SmF). Some strains that performed well in SmF processes do not perform efficiently in SSF process. Therefore, it is necessary to maximize the potential of the organism through genetic manipulation to suit the SSF process. Some of the desirable characteristics to be considered for the improvement of the microbial strains for SSF are : tolerance to low water activity and elevated temperature, deregulation of stringent metabolic controls, insensitivity to concentrations of the substrates and products. The genes responsible for counteracting the effect of increased osmotic condition, temperature, carbon and nitrogen catabolite repression are to be identified in the SSF organisms. They may be suitable manipulated in the producers or/and may be transferred to the producer organism. (Résumé d'auteur