Additional file 1 of Origin of the 3-methylglutaryl moiety in caprazamycin biosynthesis
Creators: name: Bär, Daniel
name: Konetschny, Benjamin
name: Kulik, Andreas
name: Xu, Houchao
name: Paccagnella, Davide
name: Beller, Patrick
name: Ziemert, Nadine
name: Dickschat, Jeroen S.
name: Gust, Bertolt
Publisher: name: figshare
Year: 2022
Subjects: subject: Biochemistry
subject: Space Science
subject: Microbiology
subject: FOS: Biological sciences
subject: Genetics
subject: 59999 Environmental Sciences not elsewhere classified
subject: FOS: Earth and related environmental sciences
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: Ecology
subject: Immunology
subject: FOS: Clinical medicine
subject: 69999 Biological Sciences not elsewhere classified
subject: Cancer
subject: 110309 Infectious Diseases
subject: FOS: Health sciences
subject: Plant Biology

description: Additional file 1: Figure S1. Degradation of leucine and isovalerate via the Liu-pathway as described for P. aeruginosa PAO1 [29]. Figure S2. Genetic organization of the caprazamycin biosynthetic gene cluster. Genes putatively involved in colour coding according to McErlean et al. [17]. Figure S3. Extracted ion chromatograms of S. coelicolor M1154 (three individual mutants). Masses are shown for caprazamycin aglycons E/F with m/z of 930.5, caprazamycin aglycons C/D/G with m/z of 944.5, caprazamycin aglycons A/B with m/z of 958.5 and the respective hydroxyacylcaprazols E/F with m/z of 802.5, hydroxyacylcaprazols C/D/G with m/z of 816.5 and hydroxyacylcaprazols A/B with m/z of 830.5. Figure S4. Extracted ion chromatograms of S. coelicolor M1154/cpzLK09 (three individual mutants). Masses are shown for caprazamycin aglycons E/F with m/z of 930.5, caprazamycin aglycons C/D/G with m/z of 944.5, caprazamycin aglycons A/B with m/z of 958.5 and the respective hydroxyacylcaprazols E/F with m/z of 802.5, hydroxyacylcaprazols C/D/G with m/z of 816.5 and hydroxyacylcaprazols A/B with m/z of 830.5. Figure S5. Extracted ion chromatograms of S. coelicolor M1154/cpzDB04 (three individual mutants). Masses are shown for caprazamycin aglycons E/F with m/z of 930.5, caprazamycin aglycons C/D/G with m/z of 944.5, caprazamycin aglycons A/B with m/z of 958.5 and the respective hydroxyacylcaprazols E/F with m/z of 802.5, hydroxyacylcaprazols C/D/G with m/z of 816.5 and hydroxyacylcaprazols A/B with m/z of 830.5. Figure S6. Genetic organization of clusters encoding for Liu-pathway from P. aeruginosa PAO1, caprazamycin wildtype producer S. sp. MK730-62F2 and the heterologous caprazamycin producer S. coelicolor M1154. Additional genes in Streptomyces strains are shown transparent. Table shows genes from P. aeruginosa PAO1 and their proposed function along with homologue genes found in S. sp. MK730-62F2 and S. coelicolor M1154. Values in brackets indicate % identities/similarities (n.s. no significant similarities). Figure S7. Distribution of liu clusters in Actinobacteria and Streptomyces. A: Cblaster detected 4837 similar clusters containing at least three genes homolog to the query sequence sco2775-sco2779 from S. coelicolor M1154. We discovered that 1.431 out of 31.598 (4.5%) Actinobacteria assemblies and 1.344 out of 2.574 (52.2%) Streptomyces assemblies and listed in the GTDB contain a liu cluster with homologues of all five query genes. B: Cblaster detected 5009 similar clusters containing at least three genes homolog to the query sequence sco2774-sco2779 from S. coelicolor M1154. We discovered that 1.246 out of 31.598 (3.9%) Actinobacteria assemblies and 1.241 out of 2.574 (48.2%) Streptomyces assemblies and listed in the GTDB contain a liu cluster with homologues of all six query genes. A homologue of sco2774 could only be found in 1.376 out of 5009 (27,5%) of the Actinobacteria clusters detected by cblaster, whereas the majority of detected Streptomyces cluster contain this gene with 1.343 out of 1.652 sequences (81,3%). Figure S8. Extracted ion chromatograms of S. coelicolor M1154Δsco2776-sco2779/cpzLK09 (three individual mutants). Masses are shown for caprazamycin aglycons E/F with m/z of 930.5, caprazamycin aglycons C/D/G with m/z of 944.5, caprazamycin aglycons A/B with m/z of 958.5 and the respective hydroxyacylcaprazols E/F with m/z of 802.5, hydroxyacylcaprazols C/D/G with m/z of 816.5 and hydroxyacylcaprazols A/B with m/z of 830.5. Figure S9. Extracted ion chromatograms of S. coelicolor M1154Δsco2776-sco2779/cpzDB04 (three individual mutants). Masses are shown for caprazamycin aglycons E/F with m/z of 930.5, caprazamycin aglycons C/D/G with m/z of 944.5, caprazamycin aglycons A/B with m/z of 958.5 and the respective hydroxyacylcaprazols E/F with m/z of 802.5, hydroxyacylcaprazols C/D/G with m/z of 816.5 and hydroxyacylcaprazols A/B with m/z of 830.5. Figure S10. Total ion chromatograms of S. coelicolor M1154Δsco2776-sco2779/cpzDB04 and S. coelicolor M1154Δsco2776-sco2779/cpzLK09. MS2-fragmentation patterns of peak 1 (hydoxyacylcaprazol E, Rt 12.9 min) and peak 2 (caprazamycin aglycon E, Rt 13.8 min) are shown together with corresponding fragmentation schemes. Figure S11. Extracted ion chromatograms of S. coelicolor M1154Δsco2779/cpzLK09 (three individual mutants). Masses are shown for caprazamycin aglycons E/F with m/z of 930.5, caprazamycin aglycons C/D/G with m/z of 944.5, caprazamycin aglycons A/B with m/z of 958.5 and the respective hydroxyacylcaprazols E/F with m/z of 802.5, hydroxyacylcaprazols C/D/G with m/z of 816.5 and hydroxyacylcaprazols A/B with m/z of 830.5. Figure S12. Extracted ion chromatograms of S. coelicolor M1154Δsco2779/cpzDB04 (three individual mutants). Masses are shown for caprazamycin aglycons E/F with m/z of 930.5, caprazamycin aglycons C/D/G with m/z of 944.5, caprazamycin aglycons A/B with m/z of 958.5 and the respective hydroxyacylcaprazols E/F with m/z of 802.5, hydroxyacylcaprazols C/D/G with m/z of 816.5 and hydroxyacylcaprazols A/B with m/z of 830.5. Figure S13. Extracted ion chromatograms of S. coelicolor M1154Δsco2776/cpzLK09 (three individual mutants). Masses are shown for caprazamycin aglycons E/F with m/z of 930.5, caprazamycin aglycons C/D/G with m/z of 944.5, caprazamycin aglycons A/B with m/z of 958.5 and the respective hydroxyacylcaprazols E/F with m/z of 802.5, hydroxyacylcaprazols C/D/G with m/z of 816.5 and hydroxyacylcaprazols A/B with m/z of 830.5. Figure S14. Extracted ion chromatograms of S. coelicolor M1154Δsco2776/cpzDB04 (three individual mutants). Masses are shown for caprazamycin aglycons E/F with m/z of 930.5, caprazamycin aglycons C/D/G with m/z of 944.5, caprazamycin aglycons A/B with m/z of 958.5 and the respective hydroxyacylcaprazols E/F with m/z of 802.5, hydroxyacylcaprazols C/D/G with m/z of 816.5 and hydroxyacylcaprazols A/B with m/z