Metabolome and ROAV analysis reveal the effective flavor improvement in the golden biomass of chlorophyll-deficient mutant of Auxenochlorella pyrenoidosa

The green microalga Auxenochlorella pyrenoidosa is realized a promising source of alternative protein due to its high protein content and fermentable property, but the deep green color and off-odor are the bottlenecks for its application in foods. Using atmospheric and room temperature plasma (ARTP) mutagenesis, a novel chlorophylldeficient mutant, A4-1 strain with golden color and better flavor, was successfully obtained in our previous work. In this study, to reveal the formation of the improved flavor in A4-1 strain compared to the wild type (WT), the volatiles basis was analyzed by HS-SPME Arrow-GC-MS, and the potential precursors causing flavor improvement were explored by UPLC-ESI-MS/MS. Results showed the volatiles were mainly composed of hydrocarbons, terpenoids, esters, ketones, alcohols, and aldehydes, and 29 of which were significantly different between the two biomass, largely attributed to variations of primary metabolite content, particularly those enriched in the fatty acids and amino acids. The analysis of relative odor activity value (ROAV) verified that beta-ionone, dimethyl trisulfide and (Z)-6-nonenal were the main odorants, in which the lower contributions of dimethyl trisulfide (offflavor, rancid or cooked oniony) and (Z)-6-nonenal (off-flavor, stink or fishy) were found in A4-1, which made A4-1 biomass become more neutral and superior to WT biomass in terms of odor perception. It is the first report of flavor improvement for green microalgae through breeding technology, and the novel biomass of A4-1 strain will broaden A. pyrenoidosa biomass in food application.