Fig. 2: Functional characterization of TriDTCs. | Nature Communications

Fig. 2: Functional characterization of TriDTCs.

From: Noncanonical terpene cyclases for the biosynthesis of diterpenoids regulating chlamydospore formation in plant-associated Trichoderma

Fig. 2

a GC–MS extracted ion chromatogram (EIC, m/z 272) of enzymatic assays using T. atroviride B7 protein extracts: (i) Crude protein incubated with GGPP; (ii) Purified compound 1 standard; (iii) Purified compound 2 standard; (iv) Heat-inactivated control (boiled crude protein with GGPP); (v) Negative control (crude protein without GGPP). b EIC (m/z 272) of heterologous expression products in E. coli BL21 strains containing: (i) pBbA5c-MevT-MBIS, pET-28a-triTPS4, and pCold-TF-tri4155; (ii) pBbA5c-MevT-MBIS, pCDF-Duet1-crtE, and pCold-TF-tri4155; (iii) pBbA5c-MevT-MBIS, pCDF-Duet1-crtE, and pET-28a-tri4155; and (iv) pBbA5c-MevT-MBIS and pET-28a-triTPS4. Peak in the yellow column is GGOH. c Functional analysis in A. oryzae NSAR1 systems containing: (i) PTAex3-Ptri4155-tri4155, (ii) PTAex3-PamyB-tri4155, and (iii) PTAex3 (negative control). d In vitro enzymatic tests using TriDTCs purified from E. coli system: (i) Tri4155 incubated with GGPP; (ii) Tri4155V409A/L335A with GGPP; (iii) Tcie612 with GGPP; (iv) Boiled Tri4155 with GGPP.

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