An End-to-end Pipeline for Succinic Acid Production at an Industrially Relevant Scale usingIssatchenkia orientalis

Author:

Tran Vinh G.,Mishra Somesh,Bhagwat Sarang S.,Shafaei Saman,Shen Yihui,Allen Jayne L.,Crosly Benjamin A.,Tan Shih-I,Fatma Zia,Rabinowitz Joshua,Guest Jeremy S.,Singh Vijay,Zhao Huimin

Abstract

AbstractAs one of the top value-added chemicals, succinic acid has been the focus of numerous metabolic engineering campaigns since the 1990s. However, microbial production of succinic acid at an industrially relevant scale has been hindered by high downstream processing costs arising from neutral pH fermentation. Here we describe the metabolic engineering ofIssatchenkia orientalis, a non-conventional yeast with superior tolerance to highly acidic conditions, for cost-effective succinic acid production. Through deletion of byproduct pathways, transport engineering, and expanding the substrate scope, the resulting strains could produce succinic acid at the highest titers in sugar-based media at low pH (pH 3) in fed-batch fermentations using bench-top reactors, i.e. 109.5 g/L in minimal medium and 104.6 g/L in sugarcane juice medium. We further performed batch fermentation in a pilot-scale fermenter with a scaling factor of 300×, achieving 63.1 g/L of succinic acid using sugarcane juice medium. A downstream processing comprising of two-stage vacuum distillation and crystallization enabled direct recovery of succinic acid, without further acidification of fermentation broth, with an overall yield of 64.0%. Finally, we simulated an end-to-end low-pH succinic acid production pipeline, and techno-economic analysis and life cycle assessment indicate our process is financially viable and can reduce life cycle greenhouse gas emissions by 34-90% relative to fossil-based production processes. We expectI. orientaliscan serve as a general industrial platform for the production of a wide variety of organic acids.

Publisher

Cold Spring Harbor Laboratory

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