Chaga (Inonotus obliquus) is a medicinal fungus used in traditional medicine of Native American and North Eurasian cultures. Several studies have demonstrated the medicinal properties of chaga’s bioactive molecules. For example, several terpenoids (e.g., betulin, betulinic acid and inotodiol) isolated from I. obliquus cells have proven effectiveness in treating different types of tumor cells. However, the molecular mechanisms and regulation underlying the biosynthesis of chaga terpenoids remain unknown. In this study, we report on the optimization of growing conditions for cultured I. obliquus in presence of different betulin sources (e.g., betulin or white birch bark). It was found that better results were obtained for a liquid culture pH 6.2 at 28 °C. In addition, a de novo assembly and characterization of I. obliquus transcriptome in these growth conditions using Illumina technology was performed. A total of 219,288,500 clean reads were generated, allowing for the identification of 20,072 transcripts of I. obliquus including transcripts involved in terpenoid biosynthesis. The differential expression of these genes was confirmed by quantitative-PCR. This study provides new insights on the molecular mechanisms and regulation of I. obliquus terpenoid production. It also contributes useful molecular resources for gene prediction or the development of biotechnologies for the alternative production of terpenoids.
Yellow birch barks is one of the abundant species in Quebec with harvest surplus in several regions. Biofuels or biochemicals such as biobutanol can be produced using the surplus feedstock, however challenges such as the cost of pretreatment, production of unwanted by-products in the fermentation process, and the efficient recovery of solvents must be addressed to make it feasible. The objectives of this study are to establish the optimal conditions to produce biobutanol from Eastern Canadian yellow birch; to identify natural/local Clostridium sp. strains that are capable of producing Isopropanol-Butanol-Ethanol (IBE) from synthetic sugar mixtures, as candidates for metabolic engineering and to benchmark solvent producing ability with commercially available strains; and to elucidate the challenges of paradigm shift to IBE production. Alkali pretreatment of the biomass using chemical that are present in the Kraft process were performed, followed by enzymatic hydrolysis to obtain fermentable sugars and subsequent fermentation with Clostridium acetobutylicum DSM 792. The results showed that the produced Acetone-Butanol-Ethanol (ABE) solvent concentration were 6.6-8.2 g/L of acetone; 11.2-13.1 g/L of butanol; and 2.5-2.7 g/L of ethanol. The organic acids concentration was acetic acid, 1.1-1.8 g/L, and butyric acid, 0.1-0.2 g/L. Further fermentation experiments to benchmark IBE were performed using both Clostridium beijerinckii DSM 6423 and wild isolated strains, which revealed the gaps in terms of yields and the need to optimize the fermentation paradigm. Moreover, alternative process sequences for product recovery were identified, and the impact of prior liquid-liquid extraction elucidated.
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