Additional file 1 of Development and characterization of a glycine biosensor system for fine-tuned metabolic regulation in Escherichia coli
Creators: name: Hong, Kun-Qiang
name: Zhang, Jing
name: Jin, Biao
name: Chen, Tao
name: Wang, Zhi-Wen
Publisher: name: figshare
Year: 2022
Subjects: subject: Biochemistry
subject: Microbiology
subject: FOS: Biological sciences
subject: Genetics
subject: Molecular Biology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Ecology
subject: Biological Sciences not elsewhere classified
subject: Information Systems not elsewhere classified
subject: Developmental Biology
subject: Computational Biology

description: Additional file 1: Supporting Information Supporting Information of Materials and methods. Table S1. Primers used in this study. Table S2. Plasmids used in this study. Table S3. Strains used in this study. Table S4. Glycine riboswitches used in this study. Table S5. Instant Error-prone PCR Kit instructions 2.0. Table S6. Controlled Error-prone PCR Kit instructions V1.4. Table S7. Km values of mutation and wild-type glycine riboswitches. Table S8. Sequences of synthetic glycine-ON and -OFF riboswitches. Table S9 Mutations and its locations of synthetic glycine-ON and -OFF riboswitches. Figure captions Figure S1. Glycine toxicity testing of E. coli MG1655 harboring glycine riboswitch in M9 medium with 0, 3, 7, 13, 27, 53, 80, 107, 426, 1705 mM glycine, respectively. Figure S2. Fluorescence intensity of strain MG1655-BS centrifugal pellets cultured in M9 medium supplemented with 0, 3, 7, 13, 27, 53, 80 mM glycine, respectively. Figure S3. Effect of tetA gene on glycine riboswitch and its function as genetic selection. Figure S4. Statistical analysis of mutation sites. Figure S5. Schematic diagram of long-range tertiary interactions (the α, β, γ, and δ interactions) in glycine riboswitch. Figure S6. Fermentation performances (a. OD600, b. Glucose, c. Formate, d. Acetate) of WB105-18A and WB105-18P in M9 medium supplemented with 0, 7, 13, 27, 80 mM glycine under anaerobic conditions with an initial OD600 of 0.05. Figure S7. Lactate maximum concentration of WB105-18A, WB105-15A and WB105-48A in M9 medium supplemented with 0, 1, 4, 7, 13, 27, and 80 mM glycine. Figure S8. D-Lactate production of W105-15A. Figure S9. Fermentation performances (a. Lactate, b. OD600, c. Glucose, d. Formate, e. Acetate) of WB105-15A and WB105-15P in M9 medium supplemented with 0, 7, 13, 27, 80 mM glycine under anaerobic conditions with an initial OD600 of 0.05. Figure S10. Fermentation performances (a. Lactate, b.OD600, c. Glucose, d. Formate, e. Acetate) of WB105-48A and WB105-48P in M9 medium supplemented with 0, 7, 13, 27, 80 mM glycine under anaerobic conditions with an initial OD<sub>600</sub> of 0.05. Figure S11. Statistical analysis of enrichment efficiency. Figure S12. Specific enzyme activities of 23 AGXT mutations, three AGXT library strains and control. Figure S13. Crystal structures of wild-type AGXT and mutant AGXT22. Figure S14. Distances between the site 20 in a subunit and the 43rd, 46th, 52nd, and 56th residue of another adjacent subunit on wild-type (left) and AGXT26 (right). Figure S15. Distance between the site 196 and the 7th residue of another adjacent subunit on wild-type (left) and AGXT26 (right).