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Utilization of renewable and low-cost lignocellulosic wastes has received major focus in industrial lactic acid production. The use of high solid loadings in biomass pretreatment potentially offers advantages over low solid loadings including higher lactic acid concentration with decreased production and capital costs. In this study, an isolated Enterococcus faecalis SI with optimal temperature 42 °C was used to produce optically pure L-lactic acid (> 99%) from enzyme-saccharified hydrolysates of acid-impregnated steam explosion (AISE)-treated plywood chips. The L-lactic acid production increased by 10% at 5 L scale compared to the similar fermentation scheme reported by Wee et al. The fermentation with a high solid loading of 20% and 35% (w/v) AISE-pretreated plywood chips had been successfully scaled up to process development unit scale (100 L) and pilot scale (9 m 3 ), respectively. This is the first report of pilot-scale lignocellulosic lactic acid fermentation by E. faecalis with high lactic acid titer (nearly 92 g L -1 ) and yield (0.97 kg kg -1 ). Therefore, large-scale L-lactic acid production by E. faecalis SI shows the potential application for industries.