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Figure 2 | BMC Microbiology

Figure 2

From: The hemolytic and cytolytic activities of Serratia marcescensphospholipase A (PhlA) depend on lysophospholipid production by PhlA

Figure 2

Phospholipase and hemolytic activities of S. marcescens PhlA. (A) Overnight cultures of wild-type strain S. marcescens niid 298, E. coli DH5αcells carrying pGEMeasy, E. coli DH5αcarrying pGEMeasy-phlAB, S. marcescens niid 298 phlAB deletion mutant, and S. marcescens niid 298 phlAB deletion mutant carrying pGEMeasy-phlAB (1 × 106 cells) were inoculated on blood agar plates and PCY agar plates and incubated at 37°C for 16 and 24 h, respectively. (B) Purified His-PhlA (1 μg) was separated by 12.5% SDS-PAGE, and then was stained with Coomassie blue. Protein standards were in lane M, with relative molecular masses (kDa) at the left. (C) Various phospholipids were mixed with His-PhlA and incubated at 37°C for 1 h. Free fatty acids (FFA) released from phospholipids were detected using a NEFA-C kit. The amount of FFA was determined from an oleic acid calibration curve. Values are averages ± SE of three independent experiments.

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