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. 2023 Oct 12;89(10):e01187-23. doi: 10.1128/aem.01187-23

Fig 4.

Fig 4

Characterization of the function of propane monooxygenase and the essential role of each subunit in the initial degradation of MTBE. (A) Western blot analysis of prmABCD expression in the supernatant of E. coli Rosetta (DE3) pET-prm. Lanes 1, 2, 3, and 4 represent the expression of prmA, prmB, prmC, and prmD with the C-terminal 3 × FLAG tag in the supernatant, respectively. The expected protein band sizes of PrmA-3*FLAG, PrmB-3*FLAG, PrmC-3*FLAG, and PrmD-3*FLAG were 61.6, 42.9, 14.5, and 40.0 kDa, respectively. (B) Time course of MTBE transformation and intermediate TBA production by E. coli Rosetta (DE3) (pET-prm). The TBA levels of the abiotic control and the empty vector pET-28a(+) control were below the threshold of detection. (C) Time course of TBF transformation and intermediate TBA production by E. coli Rosetta (DE3) expressing plasmid pET-prm. (D) MTBE consumption by E. coli Rosetta (DE3) containing different expression plasmids. The experiments were performed in triplicate, the results shown here are the average value of three biological replicates, and error bars show standard deviations.