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The study revealed the microbial mechanism of HNAD at high temperature and provided new insights into high-temperature biological nitrogen removal.This work aimed to firstly establish an efficient and novel two-stage cultivation process to produce microalgal biomass rich in protein using a heterotrophic Chlorella sp. MBFJNU-17 strain. In the first-stage cultivation, to reduce the glucose and urea utilization, microalga achieved a high biomass at 40 g/L glucose and 1 g/L urea; meantime, the expression from starch biosynthesis genes of micr