Anaerobic Digestion of Microalga Chlorella protothecoides and Metagenomic Analysis of Reddish-Colored Digestate

Abstract

Microalga Chlorella protothecoides materials were assessed as substrates for anaerobic digestion (AD) aiming at the simultaneous production of biogas/methane and pigments: whole autotrophic (AA) and heterotrophic algae (H); extracted heterotrophic microalgae from lipid production (HExt); and pretreated heterotrophic microalgae through enzymatic (HPEnz), autoclave (HPA), and ultrasound (HPU) processes. AA was more suitable for AD than H, as it was more efficiently converted into methane (279 vs. 180 L CH4/kg VSin). In comparison, the pretreatment of heterotrophic microalgae had a positive effect on AD, with registered methane yield increases from 263 to 290 L CH4/kg VSin (HPU, HPA, HExt). Reddish pigmentation developed in H and HPU units due to the presence of purple non-sulfur bacteria (PNSB). This phenomenon and the changes in microbiota structure during AD were confirmed by metagenomic analysis. At the end of the process, the relative abundance of Clostridiales and Bacillales increased, enhancing the hydrolysis of compounds in acetate. Consistently, Methanosaeta became the comparatively dominant methanogen, meaning that methane was produced through the acetoclastic methanogenesis pathway. The obtained results indicate for AD biorefinery feasibility—regarding the simultaneous production of biogas/methane—a digestate flow and pigments (bacteriochlorophyll a and carotenoids).