Additional file 3 of New insights into the dihydro-mureidomycin biosynthesis controlled by two unusual proteins in Streptomyces roseosporus
Creators: name: Liu, Ning
name: Xu, Yang
name: Shang, Fei
name: Sun, Huiying
name: Liu, Xiang
name: Huang, Ying
name: Tan, Huarong
name: Zhang, Jihui
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Microbiology
subject: FOS: Biological sciences
subject: Cell Biology
subject: Infectious Diseases
subject: FOS: Health sciences

description: Additional file 3. Table S1. Strains used in this study. Table S2. Plasmids used in this study. Table S3. Primers used in this study. Figure S1. Verification of SSGG-02980 and/or 03002 disruption mutants and the complementary strains. Agarose gel electrophoresis of the PCR products for verification of SSGG-02980 and/or 03002 gene disruption mutants and their complementary strains. M, DL2000 DNA marker. Lanes 1, 3, 5, 7, 9 and 11, PCR analyses of SSGG-02980 in Sr-hA, Δ02980-hA, Δ02980c-hA, Δ03002-hA, Δ03002c-hA and Δ02980/03002-hA. Lanes 2, 4, 6, 8, 10 and 12, PCR analyses of SSGG-03002 in Sr-hA, Δ02980-hA, Δ02980c-hA, Δ03002-hA, Δ03002c-hA and Δ02980/03002-hA. In the gene disruption mutants, no PCR products were obtained for the relevant genes, while the expected bands were shown in the complementory strains, verifying the correctness of the strains. Figure S2. UPLC-HRMS analyses of mureidomycins in Δ03002-hA. A, EIC spectra of MRDs (1-6) (top) and rMRDs (1’-6’) (bottom) in Δ03002-hA. Only MRDs were produced but 1’-6’ were not detected. B, Mass spectra of the extracted quasi-molecular ions of mureidomycins analogues 1-6. Figure S3. UPLC-HRMS analyses of mureidomycins in Δ02980-hA. A, EIC spectra of MRDs (1-6) (top) and rMRDs (1’-6’) (bottom) in Δ02980-hA. B, Mass spectra of the extracted quasi-molecular ions of MRDs and rMRDs (1, 3-6 and 1’, 3’-6’). The extracted MS for 2 (m/z 899.3247) and 2’ (m/z 901.3372) were not observed due to too low abundance. Compound 1’, 3’ -6’ were accumulated, while the production of 1-6 was significantly reduced. Figure S4. UPLC-HRMS analyses of mureidomycins in Δ02980c-hA. A, EIC spectra of MRDs (1-6) (top) and rMRDs (1’-6’) (bottom) in Δ02980c-hA. B, Mass spectra of the extracted quasi-molecular ions of 1-6 and 1’-6’. The production of both MRDs and rMRDs was restored. Figure S5. UPLC-HRMS analyses of mureidomycins in Δ03002c-hA. A, EIC spectra of MRDs (1-6) (top) and rMRDs (1’-6’) (bottom) in Δ03002-hA. B, Mass spectra of the extracted quasi-molecular ions of mureidomycins analogues 1, 3-6 and 1’, 3’-6’. The extracted MS for 2 (m/z 899.3245) and 2’ (m/z 901.3394) were not observed due to too low abundance. Figure S6. UPLC-HRMS analyses of mureidomycins in Δ02980/03002-hA. A, EIC spectra of MRDs (1-6) (top) and rMRDs (1’-6’) (bottom) in Δ02980/03002-hA. Only MRDs (1-6) were produced but rMRDs (1’-6’) were not detected. B, Mass spectra of the extracted quasi-molecular ions of 1-6. Table S4. NMR data for component (3) in CD3OD. a, 13C NMR data were assigned based on combination of the HMBC correlations and the comparison with that of N-acetylmureidomycin E and G. Abbreviations for the structure units: m-Tyr (m-tyrosine), AMBA (2-amino-3-methylaminobutyric acid). Table S5. NMR data for component (4) in CD3OD. a, 13C NMR data were assigned based on combination of the HMBC correlations and the comparison with that of N-acetylmureidomycin E and G. Abbreviations for the structure units: m-Tyr (m-tyrosine), AMBA (2-amino-3-methylaminobutyric acid). Table S6. NMR data for component (5) in CD3OD. a, 13C NMR data were assigned based on combination of the HMBC correlations and the comparison with that of N-acetylmureidomycin E. Abbreviations for the structure units: m-Tyr (m-tyrosine), AMBA (2-amino-3-methylaminobutyric acid). Table S7. NMR data for component (3’) in CD3OD. a, 13C NMR data were assigned based on combination of the HMBC correlations and the comparison with that of N-acetylmureidomycin E and G. Abbreviations for the structure units: m-Tyr (m-tyrosine), AMBA (2-amino-3-methylaminobutyric acid). Table S8. NMR data for component (4’) in CD3OD. a, 13C NMR data were assigned based on the combination of HMBC correlations and the comparison with that of N-acetylmureidomycin E and G. Abbreviations for the structure units: m-Tyr (m-tyrosine), AMBA (2-amino-3-methylaminobutyric acid). Figure S7. 1H NMR spectrum (500 MHz) of component (3) in CD3OD. Figure S8. 13C NMR spectrum (125 MHz) of component (3) in CD3OD. Figure S9. Dept135 spectrum of component (3) in CD3OD. Figure S10. COSY spectrum of component (3) in CD3OD. Figure S11. HSQC spectrum of component (3) in CD3OD. Figure S12. HMBC spectrum of component (3) in CD3OD. Figure S13. 1H NMR spectrum (500 MHz) of component (4) in CD3OD. Figure S14. 13C NMR spectrum (125 MHz) of component (4) in CD3OD. Figure S15. Dept135 spectrum of component (4) in CD3OD. Figure S16. COSY spectrum of component (4) in CD3OD. Figure S17. HSQC spectrum of component (4) in CD3OD. Figure S18. HMBC spectrum of component (4) in CD3OD. Figure S19. 1H NMR spectrum (500 MHz) of component (5) in CD3OD. Figure S20. 13C NMR spectrum (125 MHz) of component (5) in CD3OD. Figure S21. Dept135 spectrum of component (5) in CD3OD. Figure S22. COSY spectrum of component (5) in CD3OD. Figure S23. HSQC spectrum of component (5) in CD3OD. Figure S24. HMBC spectrum of component (5) in CD3OD. Figure S25. 1H NMR spectrum (500 MHz) of component (3’) in CD3OD. Figure S26. 13C NMR spectrum (125 MHz) of component (3’) in CD3OD. Figure S27. Dept135 spectrum of component (3’) in CD3OD. Figure S28. COSY spectrum of component (3’) in CD3OD. Figure S29. HSQC spectrum of component (3’) in CD3OD. Figure S30. HMBC spectrum of component (3’) in CD3OD. Figure S31. 1H NMR spectrum (500 MHz) of component (4’) in CD3OD. Figure S32. 13C NMR spectrum (125 MHz) of component (4’) in CD3OD. Figure S33. Dept135 spectrum of component (4’) in CD3OD. Figure S34. COSY spectrum of component (4’) in CD3OD. Figure S35. HSQC spectrum of component (4’) in CD3OD. Figure S36. HMBC spectrum of component (4’) in CD3OD. Figure S37. 1H NMR spectrum (500 MHz) of component (5’) in CD3OD. Figure S38. 13C NMR spectrum (125 MHz) of component (5’) in CD3OD. Figure S39. Putative structure of compound 2’ and its isomer.