Rice straw-derived lipid production by HMF/furfural-tolerant oleaginous yeast generated by adaptive laboratory evolution.

Research on producing medium- and long-chain hydrocarbons as drop-in biofuels has recently accelerated. In addition, lipids are emerging as precursors for biofuel production, and thus, microbial lipid production utilizing agrowastes is becoming a feasible platform technology. Nonetheless, microorganisms are often inhibited by furan aldehydes in biomass-derived hydrolysates. Accordingly, this study aimed to develop oleaginous yeast strains that can tolerate furan aldehydes for producing lipids as biofuel precursors. Rhodosporidium toruloides was selected as the target for adaptive laboratory evolution. The evolved strain, which was obtained from 16 rounds of subcultures, showed a 2.5-fold higher specific growth rate than the wild-type strain in the presence of furan aldehydes and slightly higher lipid production in rice straw hydrolysate. The results discussed in this study provide insights into the production of lipid production by oleaginous yeast utilizing agrowastes as feedstock to obtain drop-in biofuels and contribute to feasible strategies to address climate crises.

Adsorption Study of Acid Soluble Lignin Removal from Sugarcane Bagasse Hydrolysate by a Self-Synthesized Resin for Lipid Production.

An adsorption resin CX-6 was synthesized and used for acid soluble lignin (ASL) removal from sugarcane bagasse hydrolysate (SCBH). The adsorption conditions of pH value, amount of adsorbent, initial ASL concentration, and temperature on ASL adsorption were discussed. The results showed the adsorption capacity of ASL was negatively affected by increasing temperature, solution pH, and adsorbent dose, and was positively affected by increasing initial concentration. The maximum adsorption capacity of ASL was 135.3 mg/g at initial ASL concentration 6.46 g/L, adsorption temperature 298 K, and pH 1. Thermodynamic study demonstrated that the adsorption process was spontaneous and exothermic. Equilibrium and kinetics experiments were proved to fit the Freundlich isotherm model and pseudo-second-order model well, respectively. Fermentation experiment showed that the SCBH after combined overliming with resin adsorption as fermentation substrate for microbial lipid production by Trichosporon cutaneum and Trichosporon coremiiforme was as better as that of SCBH by combined overliming with active charcoal adsorption, and more efficient than that of SCBH only by overliming. Moreover, the regeneration experiment indicated that the CX-6 resin is easy to regenerate and its recirculated performance is stable. In conclusion, our results provide a promising adsorbent to detoxify lignocellulose hydrolysate for further fermentation.

Use of Crude Glycerol as Sole Carbon Source for Microbial Lipid Production by Oleaginous Yeasts.

Crude glycerol, discharged from biodiesel production process, is a potential carbon source for microbial lipid production. The capability of using crude glycerol as sole carbon source for microbial lipid production by oleaginous yeasts Trichosporon fermentans and Trichosporon cutaneum was investigated for the first time. T. fermentans and T. cutaneum could use crude glycerol for efficient lipid production, and the optimal glycerol concentration for which were 50 and 70 g/L, respectively. The optimum nitrogen source, C/N, inoculum concentration, and pH were yeast extract + peptone, 60, 10.0%, and 6.0, respectively. The most suitable culture temperature for T. fermentans and T. cutaneum were 28 and 30 °C, respectively. Under the optimal conditions, the maximum biomass, lipid content, lipid yield, and lipid coefficient of T. fermentans and T. cutaneum were 16.0 g/L, 32.4%, 5.2 g/L, and 16.5%, and 17.4 g/L, 32.2%, 5.6 g/L, and 17.0%, respectively. Moreover, it was found that methanol present in the crude glycerol had minor influence on the lipid production. Addition of surfactant potassium oleate into the medium could slightly stimulate the cell growth and lipid accumulation of both yeasts. This study shows that T. fermentans and T. cutaneum are promising strains for lipid production on crude glycerol.

Characterization of newly isolated oleaginous yeasts - Cryptococcus podzolicus, Trichosporon porosum and Pichia segobiensis.

The yeast strains Cryptococcus podzolicus, Trichosporon porosum and Pichia segobiensis were isolated from soil samples and identified as oleaginous yeast strains beneficial for the establishment of microbial production processes for sustainable lipid production suitable for several industrial applications. When cultured in bioreactors with glucose as the sole carbon source C. podzolicus yielded 31.8% lipid per dry biomass at 20°C, while T. porosum yielded 34.1% at 25°C and P. segobiensis 24.6% at 25°C. These amounts correspond to lipid concentrations of 17.97 g/L, 17.02 g/L and 12.7 g/L and volumetric productivities of 0.09 g/Lh, 0.1 g/Lh and 0.07 g/Lh, respectively. During the culture of C. podzolicus 30 g/l gluconic acid was detected as by-product in the culture broth and 12 g/L gluconic acid in T. porosum culture. The production of gluconic acid was eliminated for both strains when glucose was substituted by xylose as the carbon source. Using xylose lipid yields were 11.1 g/L and 13.9 g/L, corresponding to 26.8% and 33.4% lipid per dry biomass and a volumetric productivity of 0.07 g/Lh and 0.09 g/Lh, for C. podzolicus and T. porosum respectively. The fatty acid profile analysis showed that oleic acid was the main component (39.6 to 59.4%) in all three strains and could be applicable for biodiesel production. Palmitic acid (18.4 to 21.1%) and linolenic acid (7.5 to 18.7%) are valuable for cosmetic applications. P. segobiensis had a considerable amount of palmitoleic acid (16% content) and may be suitable for medical applications.