Microwave-assisted enzymatic extraction (MAEE) was used for the separation of polysaccharides from micro-Chlorella. The extraction condition of MAEE was optimized by Box-Behnken design and response surface methodology. Results showed that the optimal condition for the extraction of Chlorella sp. crude polysaccharides (CSCP) was at 50°C for 2.3 h with 380 W of microwave power and 0.31% of enzyme dosage. Under the optimal extraction condition, the extraction yield of CSCP reached 0.72%. Similarly, the α-amylase modification conditions of the CSCP were also optimized, in which the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging rate was used as the response value. The scavenging rate of DPPH free radicals was 17.58% when enzyme dosage was 271 U/g at 51°C for 14 min. Moreover, the enzyme-modified CSCP presented a typical heteropolysaccharide mainly including glucose (48.84%), ribose (13.57%) and mannose (11.30%). MAEE used in this work achieved a high extraction yield of CSCP, which provides an efficient method for the extraction of CSCP from Chlorella sp.

Digestates from different anaerobic digesters are promising substrates for microalgal culture, leading to effective wastewater treatment and the production of microalgal biomass. However, further detailed research is needed before they can be used on a large scale. The aims of this study were to investigate the culture of Chlorella sp. in Digestate_M from anaerobic fermentation of brewer’s grains and brewery wastewater (BWW) and to explore the potential use of the biomass produced under different experimental conditions, including diverse cultivation modes and dilution ratios. Cultivation in Digestate_M initiated from 10% (v/v) loading, with 20% BWW, obtained maximum biomass production, reaching 1.36 g L⁻¹ that was 0.27g L⁻¹ higher than 1.09 g L⁻¹ of BG11. In terms of Digestate_M remediation, the maximum removal of ammonia nitrogen (NH₄⁺-N), chemical oxygen demand, total nitrogen, and total phosphorus reached 98.20%, 89.98%, 86.98%, and 71.86%, respectively. The maximum lipid, carbohydrate, and protein contents were 41.60%, 32.44%, and 27.72%, respectively. The growth of Chlorella sp. may be inhibited when the Y(II)–F_v/F_m ratio is less than 0.4.

Androstenedione (ADSD) is one of the widely detected androgens in diverse aquatic environments. However, there were few reports on the molecular mechanism of Chlorella vulgaris exposure to ADSD. In our previous research, we have investigated the genes associated with chlorophyll metabolism in Chlorella vulgaris response to ADSD. In this study, we focus on continuously up-regulated genes to explore the mechanism underlying Chlorella vulgaris resistance to ADSD toxicity. Chlorella vulgaris was exposed to ADSD with five concentration gradients. The continuously up-regulated genes were enriched by Series Test of Cluster (STC) analysis and verified by qRT-PCR. Microalgae Super Oxidase Dimutase (SOD) and Microalgae Malonic dialdehyde (MDA), two indicators of oxidative stress, were determined by ELISA after exposure to ADSD. The results showed that ADSD can stimulate the production of extracellular polymeric substances (EPS) and lead to enlargement in the cell body of Chlorella vulgaris. In addition, steroid biosynthesis and oxidoreductase activity processes were consistently up-regulated upon exposure to ADSD. In conclusion, our study highlighted the crucial role of phenotypic modification, hormone synthesis, and redox mechanisms in protecting Chlorella vulgaris cells from the harmful effects of ADSD contamination.