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During the conversion of lignocellulose, phenolic wastewaters are generated. Therefore, researchers have investigated wastewater valorization processes in which these pollutants are converted to chemicals, i.e., lipids. However, wastewaters are lean feedstocks, so these valorization processes in research typically require the addition of large quantities of sugars and sterilization, which increase costs. This paper investigates a repeated batch fermentation strategy with Rhodotorula kratochvilovae EXF7516 and Cutaneotrichosporon oleaginosum ATCC 20509, without these requirements. The pollutant removal and its conversion to microbial oil were evaluated. Because of the presence of non-monomeric substrates, the ligninolytic enzyme activity was also investigated. The repeated batch fermentation strategy was successful, as more lipids accumulated every cycle, up to a total of 5.4 g/L (23% cell dry weight). In addition, the yeasts consumed up to 87% of monomeric substrates, i.e., sugars, aromatics, and organics acids, and up to 23% of non-monomeric substrates, i.e., partially degraded xylan, lignin, cellulose. Interestingly, lipid production was only observed during the harvest phase of each cycle, as the cells experienced stress, possibly due to oxygen limitation. This work presents the first results on the feasibility of valorizing non-sterilized lignocellulosic wastewater with R. kratochvilovae and C. oleaginosum using a cost-effective repeated batch strategy.

Microalgae contain valuable compounds that can be harnessed for industrial applications. Lignocellulose biomass is a plant material containing in abundance organic substances such as carbohydrates, phenolics, organic acids and other secondary compounds. As growth of microalgae on organic substances was confirmed during heterotrophic and mixotrophic cultivation, lignocellulose derived compounds can become a feedstock to cultivate microalgae and produce target compounds. In this review, different treatment methods to hydrolyse lignocellulose into organic substrates are presented first. Secondly, the effect of lignocellulosic hydrolysates, organic substances typically present in lignocellulosic hydrolysates, as well as minor co-products, on growth and accumulation of target compounds in microalgae cultures is described. Finally, the possibilities of using lignocellulose hydrolysates as a common feedstock for microalgae cultures are evaluated.

This work evaluates the possibility of applying enzymatic beech wood (Fagus sylvatica) hydrolysate as a feedstock for Chlorella sorokiniana growth, and fatty acid and pigment production. Beech wood solids were pretreated with NaOH at high temperature to partially remove xylose and Klason lignin, and enable production of glucose during subsequent enzymatic hydrolysis. Neutralized wood enzymatic hydrolysate containing glucose (TGP-Enz10), was tested on Chlorella growth during heterotrophic cultivation and compared with microalgae growth in a medium containing synthetic glucose (TGP). Results show that enzymatic hydrolysate enabled Chlorella growth in the dark for biomass, fatty acid and pigment production due to the presence of glucose, although the productivity obtained was smaller, if compared to heterotrophic cultivation in a synthetic TGP medium. Partial growth inhibition and diminished productivity in wood hydrolysate supplemented Chlorella culture was due to the presence of neutralized citrate buffer. Neutralized citrate buffer (TGP-Cit10) was found to partially inhibit heterotrophic growth and also strongly suppress mixotrophic growth in Chlorella culture. This buffer was also shown to alter fatty acid composition and to slightly affect ChlTotal/CarTotal ratio during heterotrophic cultivation. Heterotrophic Chlorella cultivation with TGP-Enz10 showed that wood enzymatic hydrolysate can constitute a potential feedstock for microalgae cultivation, although the composition of the buffer used during enzymatic hydrolysis should be taken into consideration.