Browsing by Author "Marapara, Jorge L."

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A simple and efficient method for high-quality total RNA isolation from oleaginous microalgae
(Plant Cell Biotechnology and Molecular Biology, 18(1-2), 15-21, 2017-01-06) Castro, Juan C.; Rodríguez, Hicler N.; Maddox, J. Dylan; Jiu, Bruce; Petterman, Joe B.; Marapara, Jorge L.; Cobos, Marianela
Microalgae are a very diverse group of microorganisms with great potential for biotechnological applications. To best exploit these organisms is necessary to expand our knowledge at its molecular levels. To realize these studies, however, it is essential to have efficient and reproducible methods for total RNA isolation. The objective of this research was to develop a method for high-quality total RNA isolation from three oleaginous microalgae species promissories for biotechnological applications. Three strains Ankistrodesmus sp., Chlorella sp., and Scenedesmus sp. were cultured on CHU-10 medium. Then, were harvested by centrifugation and total RNA was isolated with three commercial kits, one previously published method, and our improved CTAB-LiCl method. Quality and quantity of total RNA isolated were assayed with standard spectrophotometric and electrophoretic techniques. The highest quality (A260/A280: 1.95-2.02, A260/A230: 1.12-1.84) and yield (16.40-93.63 mg/g fresh weight) of total RNA from the three microalgae strains were obtained with our improved CTAB-LiCl method. In addition, the time required to complete the isolation process and the cost were relatively lowest. In conclusion, the improved CTAB-LiCl method, in contrast to frequently used commercial kits, and a previously published method, is effective and reliable to produce total RNA of high quality and quantity that is suitable for cDNA synthesis and polymerase chain reaction.
De novo RNA-Seq analysis of the oleauginous microalgae Ankistrodesmus sp. UCP0001: Gene identification and metabolic pathways reconstruction for the biosynthesis of fatty acids and triacylglycerols
(Plant Cell Biotechnology and Molecular Biology 18(5-6): 219-230, 2017-05-15) Castro, Juan C.; Maddox, J. Dylan; Paredes, Jae D.; Rodríguez, Hicler N.; Aguilar, Carla P.; Marapara, Jorge L.; Castro, Carlos G.; Casuso, María Z.; Cobos, Marianela
Microalgae have great potential as feedstock to produce next-generation biofuels. The scarceness of genomic level information, however, prevents the rational de novo microalgae strain design. In this research, we describe the next-generation sequencing, de novo assembly, and functional annotation of the transcriptome of Ankistrodesmus sp. UCP0001. In total 48,867,830 high-quality sequence reads were de novo assembled into 38,414 unigenes (mean length = 508 bp, N50 = 1038 bp). Seventy-two percent of unigenes presented mapping information. Based on the KEGG pathway assignment, the fatty acids and the triacylglycerol biosynthesis pathways were reconstructed. Our results demonstrate that the synergy among high-throughput sequencing technologies and appropriate bioinformatics tools provides a fast, low-cost, and effective approach to generate invaluable functional genomic information in non-model microalgae species (e.g., Ankistrodesmus sp.). With the de novo assembled and annotated transcriptome we have successfully identified genes encoding enzymes and reconstructed metabolic pathways for the biosynthesis of fatty acids and triacylglycerols in this microalgae species. This genetic information could be used for the de novo microalgae strain design with desirable characteristics to produce biodiesel and capabilities for the biosynthesis of others valuable bioactive compounds of interest to the pharmacological, food, and cosmetic industries.
Isolation, Characterization, and Biotechnological Potential of Native Microalgae From the Peruvian Amazon
(Universidad Científica del Perú, 2019-08-31) Cobos Ruiz, Marianela; Castro Gómez, Juan C.; Paredes Rodríguez, Jae D.; Pérez, Sheyla; Dylan Maddox, Joseph; Estela Moreno, Segundo L; Tresierra Ayala, Álvaro Benjamín; Saldaña Rojas, Juan R.; Marapara, Jorge L.; Adrianzén, Pedró M.; Gonzáles, Rosana
The objective of this chapter is to provide scientific information on basic aspects to be taken into account to achieve the successful isolation, biochemical, and molecular characterization and then to evaluate the biotechnological potential of native microalgae of the Peruvian Amazon. Recent investigations reported by our research team has demonstrated that the isolated native microalgae from the Peruvian Amazon have a great potential for the biotechnological production of biodiesel and nutraceuticals. This biotechnological potential was identified thanks to the application of various protocols that were standardized by the authors over the last 5 years. In conclusion, the native microalgae of the Peruvian Amazon have biotechnological potential and are therefore promising for the production of both biodiesel and nutraceuticals. Various species of microalgae were identified, isolated, cultured, and characterized using biochemical, nutraceutical, and molecular techniques, the isolation stage being the starting point to achieve various biotechnological applications. Ankistrodesmus sp. is one of the microalgae with potential for the production of biodiesel and microalgae such as Haematococcus pluvialis, Scenedesmus sp., and Chlorella sp., among others demonstrated a high potential for nutraceutical production. The stress conditions to which microalgae are subjected are being a determining factor for the production of biodiesel and nutraceuticals.
Nutritional evaluation and human health-promoting potential of compounds biosynthesized by native microalgae from the Peruvian Amazon
(Universidad Científica del Perú, 2020-07-12) Cobos Ruiz, Marianela; Pérez, Sheyla; Braga, Janeth; Vargas Arana, Gabriel; Flores, Leenin; Paredes Rodríguez, Jae Diana; Dylan Maddox, Joseph; Marapara, Jorge L.; Castro Gómez, Juan C.
A plausible strategy to mitigate socioeconomic problems in the Peruvian Amazon is through the sustainable exploitation of biodiversity resources, such as native microalgae. Several studies worldwide affirm that these microorganisms are excellent sources of higher value products for human nutrition and possess health-promoting biochemicals, but these attributes are unknown for the native microalgae of Peru. Therefore, the aim of this investigation was to evaluate the nutritional and human health-promoting potential of compounds biosynthesized by native microalgae from the Peruvian Amazon. Ten native microalgae strains of the groups cyanobacteria and chlorophyta were cultured in BG-11 medium and their biomass harvested and dried. Standardized methods were then used to determine proximate composition, fatty acids and amino acids composition, antioxidant activity, and total phenolic content. All ten microalgae strains produce primary nutrients, the entire spectrum of essential amino acids, essential fatty acids, and 3 of the 10 microalgae strains produced eisosapentaenoic acid. Additionally, all microalgae strains exhibited antioxidant activities and contained phenolic compounds. In conclusion, native microalgae strains from the Peruvian Amazon analyzed in this study possess the ability to biosynthesize and accumulate several nutrients and compounds with human health-promoting potential.