Production of biofuels and fine chemicals from biomass-derived carbohydrates through biorefinery attracts much attention because it is recognized as an environmentally friendly process. Microalgae can serve as promising carbohydrate producers for biorefinery rather than woody and crop biomass due to high biomass productivity, high CO2 fixation, and no competition with food production. However, microalgae with high carbohydrate productivity have not been well investigated despite intensive studies of microalgal lipid production. In this study, the carbohydrate production of Pseudoneochloris sp. strain NKY372003 isolated as a high carbohydrate producer, was investigated. Cultivation conditions with various combinations of nutrient contents and photon flux density were examined to maximize the biomass and carbohydrate productivities. At the optimal condition, the biomass and carbohydrate production of this strain reached 8.11 +/- 0.37 g/L and 5.5 +/- 0.2 g/L, respectively. As far as we know, this is the highest carbohydrate production by microalgae among ever reported. Cell staining with Lugol's solution visualized intracellular starch granules. Because algal starch can be converted to biofuels and building blocks of fine chemicals, Pseudoneochloris sp. NKY372003 will be a promising candidate for production of fermentable carbohydrates towards biofuels and fine chemicals production. (C) 2020, The Society for Biotechnology, Japan. All rights reserved.