Additional file 1 of Production of extracellular amylase contributes to the colonization of Bacillus cereus 0–9 in wheat roots
Creators: name: Huang, Qiubin
name: Liu, Huiping
name: Zhang, Juanmei
name: Wang, Shaowei
name: Liu, Fengying
name: Li, Chengdie
name: Wang, Gang
Publisher: name: figshare
Year: 2022
Subjects: subject: Microbiology
subject: FOS: Biological sciences
subject: Genetics

description: Additional file 1: Fig. S1. Analysisthe amyC gene (FRY47_05825) signal peptide and smart domain 4 (A) Analysis theAmyC protein signal peptide through SignalP 4.1. S-score: Each 5 amino acidcorresponds to 1 S value, with a high S value in the signal peptide region. 6C-score: Each amino acid will have a C value, with the highest C value at theshear 7 site. Y-score: Y value is a parameter considering S value and C value,which is 8 accurate than C value alone, because there may be more than onehigher C value in a 9 series, but only one shear site; the shear site is withsteep S value and high C value. 10 (B) Analysis the AmyC protein smart domainthrough SMART software. The 60-330 11 position is catalytic activity, and thereare two transmembrane areas at each end of the 12 active area. Fig. S2. Analysis the amyP gene(FRY47_27030) signal peptide and smart domain. 15 (A) Analysis the AmyP proteinsignal peptide through SignalP 4.1. S-score: Each 16 amino acid corresponds to1 S value, with a high S value in the signal peptide region. 17 C-score: Eachamino acid will have a C value, with the highest C value at the shear 18 site.Y-score: Y value is a parameter considering S value and C value, which is 19accurate than C value alone, because there may be more than one higher C valuein a 20 series, but only one shear site; the shear site is with steep S valueand high C value. 21 (B) Analysis the AmyP protein smart domain through SMARTsoftware. The 23 to 22 401 is the catalytic activity, 413 to 496 is the Cdomain of Aamy, 501 to 577 is the 23 TIG domain, 585 to 681 is the domain ofCBM 20, which has starch binding function. Fig.S3. No strains colonization on wheat root (control treatment). Fig. S4. Induced expression of amySgene in E.coli BL21(DE3) strain. The strain was cultured to 10 5 31 cfu/mL andsupplemented with 100 μmol/L IPTG for induction culture 32 at 22°C for 6 h.Then subsequently centrifugated and subjected to ultrasonication. 20 33 mM imidazolesolution was used for a Ni column purification, and obtained the 34 purifiedprotein. (a) represent the commercial α-amylase(Ruibio, Germany); (b) 35represent unpurified AmyS protein; (c) represent purified AmyS protein; (d)represent 36 the purified protein was diluted twice-fold; (e) represent thepurified protein was 37 diluted three-fold; (f) represent the purified proteinwas diluted four-fold; (g) 38 represent the purified protein was dilutedfive-fold; (h) represent the purified protein 39 was diluted six-fold. Fig. S5. Determination of theinhibitory effect of bacteria and mutant strains on R. 42 cerealis on agarplates. The strains were inoculated in liquid LB medium, cultured at 30 °C to10 8 43 cfu/mL, collected, and washed twice with PBS. Then, an equal volume of44 sterile water was added to the pellet, and 5 μL of bacterial suspension washarvested, 45 inoculated on PDA solid medium, cultured at 30 °C for 3 d. Arepresent the strain 0-9; 46 B represent the strain ∆amyS ; C represent the strain∆amySec; D represent the strain 47 ∆amyS/amyS;, Table S1. The primers used in this study.