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. 2022 Jun 7;50(11):6587–6600. doi: 10.1093/nar/gkac476

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© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 5. — Improving 2′-fucosyllactose production by ordered control of competitive pathways and enhancing GTP supply. (A) A bifunctional thermosensitive genetic circuit was constructed using the temperature-responsive biosensors and CRISPRi based NOT gate. The 2′-fucosyllactose synthesis and GTP supply were upregulated by the temperature-responsive biosensor, and the competitive pathways were downregulated by the temperature -responsive repression cascade. G6P, glucose-6-phosphate; F6P, fructose-6-phosphate; M6P, mannose-6- phosphate; M1P, mannose-1-phosphate. (B) Effects of weakening PPP and glycolysis on 2′-fucosyllactose production. (C) Trends of cell growth, lactose, sucrose, extracellular and intracellular 2′-fucosyllactose concentration of the strain MT9 under ordered control from 30 to 34 to 37°C (changing the temperature to 34 and 37°C orderly after 12- and 18-h growth). (D) The metabolic pathway of GTP synthesis in B. subtilis. (E) The cell growth and titer of GTP recycling-modified strains. In addition, trends of cell growth, sucrose, extracellular and intracellular 2′-fucosyllactose concentration of the strain MT17 are shown. Data are presented as mean ± s.d. of three replicates.