Data for Xylose Metabolic Engineering of Issatchenkia orientalis for 3-Hydroxypropionic Acid Production from Cellulosic Hydrolysates without Nutrient Supplementation

Tan, L.R., Kim, S.M., Cho, Y.B., Tan, S.I., Deshavath, N.N., Wei, N., Singh, V., Yoshikuni, Y., Zhao, H., Jin, Y.S. May 3, 2026. Data for: “Xylose Metabolic Engineering of Issatchenkia orientalis for 3-Hydroxypropionic Acid Production from Cellulosic Hydrolysates without Nutrient Supplementation.” University of Illinois Urbana-Champaign. DOI: 10.13012/B2IDB-1480828_V1.

Bioconversion of lignocellulosic biomass offers a promising alternative to petroleum-based chemical production. However, inefficient xylose utilization and toxic compounds in cellulosic hydrolysate limit microbial fermentation, as the hydrolysate contains substantial amounts of xylose in addition to glucose. To address these challenges, we engineered Issatchenkia orientalis to produce 3-hydroxypropionic acid (3-HP) directly from sorghum hydrolysate under low-pH conditions. A heterologous xylose utilization pathway consisting of XYL1, XYL2, and XYL3 from Scheffersomyces stipitis was introduced into an engineered 3-HP producing strain, enabling efficient conversion of xylose to 3-HP. The engineered strain produced 46.8 g/L 3-HP from sorghum hydrolysate without nutrient supplementation. To eliminate the lag phase under low-pH conditions, fermentation was conducted at pH 6.0 for the first three days, after which pH control was discontinued and in situ 3-HP accumulation buffered the culture. This partial pH control strategy increased 3-HP productivity by 55% from 0.20 to 0.31 g/L•h, while maintaining low-pH conditions. Introducing an additional copy of XYL2 further increased 3-HP titer to 53.5 g/L and the yield by 33%, from 0.30 to 0.40 g/g sugars, with pH reaching 4.5 at the end of fermentation. This represents one of the highest reported 3-HP titers and yields from cellulosic hydrolysate without additional nutrient supplementation. This work demonstrates a nutrient-independent and low-pH bioprocess for upgrading lignocellulosic hydrolysate into 3-HP, highlighting the industrial potential of engineered xylose-utilizing I. orientalis for sustainable production of platform chemicals from renewable feedstocks.

Illinois Data Bank: Plasmids, primers, 3-HP/xylitol/fermentation profiles, pH changes