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Fig. 1 | Microbial Cell Factories

Fig. 1

From: Functionalized PHB granules provide the basis for the efficient side-chain cleavage of cholesterol and analogs in recombinant Bacillus megaterium

Fig. 1

Characterization of B. megaterium (strain MS941) whole-cell system for the conversion of cholesterol and analogs. a Tricistronic vector pSMF2.1_SCCAA encoding side-chain cleavage enzyme CYP11A1 as well as redox partners AdR and Adx under the control of a xylose-inducible promoter (P xylA , promoter xylose utilization operon; xylR, xylose repressor; tetR, tetracycline resistance gene; ampR, ampicillin resistance gene). b HPLC-chromatograms of the conversion of cholesterol after 48 h (light gray strain MS941 transformed with pSMF2.1_SCCAA, black untransformed strain, S cholesterol, P pregnenolone, F flow-through peak). c HPLC-chromatograms of the conversion of 7-DHC after 48 h (light gray strain MS941 transformed with pSMF2.1_SCCAA, black untransformed strain, S 7-DHC, P 7-DHP, F flow-through peak). d HPLC chromatograms of the conversion of β-sitosterol after 48 h (light gray strain MS941 transformed with pSMF2.1_SCCAA, black untransformed strain, S β-sitosterol, P pregnenolone, C campesterol contained in the sample as minor impurity, F flow-through peak). e Effect of different final concentrations of Quillaja saponin (inset) on product yield. The substrate was dissolved in a 45% 2-hydroxypropyl-β-cyclodextrin with varying saponin concentrations and added to the cultures directly after protein induction. Pregnenolone yield after cholesterol conversion was determined by HPLC. f Conversion rates of substrates cholesterol, 7-DHC and β-sitosterol after 48 h. Error bars indicate standard deviation of triplicate biological experiments.

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