Enhancing synthesis of ethyl lactate in rice baijiu fermentation by adding recovered granular cells

Ethyl lactate is the most abundant ester in semi-solid rice baijiu fermentation, affecting the flavor of baijiu to a great extent. The present study aimed to investigate the spatial distribution and formation contributor of ethyl lactate by removing the microorganisms and extracellular enzymes from the upper, middle, and lower fermentation broth during the later fermentation stage. The removal of suspended substances by centrifugation did not affect the ethyl lactate content in the top and middle fermentation broth containing free cells, enzymes, and starch particles. After day 5 of fermentation, only the lower fermentation broth containing granular cells attached to the starch could continue to accumulate lactic acid, thereby increasing the ethyl lactate content. The results showed that the chemical reactions were the main contributor to the increased ethyl lactate content at the anaphase of fermentation rather than enzymatic catalysis or microbial metabolism. Sequencing of granular cells revealed the main lactic acid producers at different fermentation stages. Lactobacillus helveticus showed the highest abundance of 94.45-95.40% on day 5, which decreased to 29.58-30.20% on day 15, while Lactobacillus acetotolerans showed the highest abundance of 47.93-49.72% at day 15. Additionally, the granular cells were recovered and used for supplementary inoculation in the next batch, which significantly increased the ethyl lactate content. This study provided a novel strategy for improving the ethyl lactate content in semi-solid baijiu fermentation.