Abstract
Microbial biorefinery is a promising route toward sustainable production of glycolic acid (GA), a valuable raw material for various industries. However, inherent microbial GA production has limited substrate consumption using either d-xylose or d-glucose as carbon catabolite repression (CCR) averts their co-utilization. To bypass CCR, a GA-producing strain using d-xylose via Dahms pathway was engineered to allow cellobiose uptake. Unlike glucose, cellobiose was assimilated and intracellularly degraded without repressing d-xylose uptake. The final GA-producing E. coli strain (CLGA8) has an overexpressed cellobiose phosphorylase (cep94A) from Saccharophagus degradans 2–40 and an activated glyoxylate shunt pathway. Expression of cep94A improved GA production reaching the maximum theoretical yield (0.51 g GA g−1 xylose), whereas activation of glyoxylate shunt pathway enabled GA production from cellobiose, which further increased the GA titer (2.25 g GA L−1). To date, this is the highest reported GA yield from d-xylose through Dahms pathway in an engineered E. coli with cellobiose as co-substrate.
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Acknowledgements
The authors would like to acknowledge the National Research Foundation of Korea (NRF) under the Basic Science Research Program through the Ministry of Education and the Korea Institute of Technology Evaluation and Planning (KETEP) funded by the Ministry of Trade, Industry & Energy for funding the research.
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This work was supported by the National Research Foundation of Korea (NRF) under the Basic Science Research Program through the Ministry of Education (2018R1D1A1B07043993 and 2020R1A6A1A03038817) and by the Korea Institute of Technology Evaluation and Planning (KETEP) funded by the Ministry of Trade, Industry & Energy (MOTIE No. 20194010201750).
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RBC and ABB conceived and designed the study, performed the experiments, analyzed the data, and wrote the manuscript. GMN analyzed the data, supervised, wrote, and revised the manuscript. WKL and WJC acquired the funding and resources, supervised, and revised the manuscript. All the authors read and approved the final version of the manuscript.
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Cabulong, R.B., Bañares, A.B., Nisola, G.M. et al. Enhanced glycolic acid yield through xylose and cellobiose utilization by metabolically engineered Escherichia coli. Bioprocess Biosyst Eng 44, 1081–1091 (2021). https://doi.org/10.1007/s00449-020-02502-6
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DOI: https://doi.org/10.1007/s00449-020-02502-6