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. 2023 Jan 6;33(3):319–328. doi: 10.4014/jmb.2212.12026

Fig. 4. Identification of mainly expressed secretory aspartyl proteases in M. restricta and the inhibitory effect of M. restricta on the formation of S. aureus biofilm.

Fig. 4

A. Unrooted phylogenetic tree of the putative secretory aspartyl proteases in M. restricta compared with MGL_1932 (MgSAP1) and MGL_3328 in M. globosa and FUN_000223 (MfSAP1) in M. furfur. The numbers at the nodes indicate the bootstrap values. B. Coomassie brilliant blue staining of SDSPAGE of the extracellular media (Flow though), binding buffer (Binding), and the enriched fraction (Elution) from pepstatin A-agarose affinity purification. The contents of the band in the block box (approximately 42 kDa) were analyzed by orbitrap mass spectrometry and identified as MRET_3770, MRET_2827, MRET_2825, and MRET_2661. C. Biofilm enrichment was measured by the crystal violet assay as described in the Methods section. Serial concentration of S. aureus (S.A) in axenic culture, S.A co-cultured with M. restricta (M.R), or S.A cultured in M.R spent media were examined. Black box, S.A axenic culture; red box, S.A co-cultured with M.R (OD600: 1); blue box, S.A co-cultured with M.R (OD600: 0.1); green box, S.A cultured in M.R spent media; yellow box, M.R axenic culture. D. The ratios of biofilm formation. Biofilm formation was measured by spectrophotometry. *, p < 0.1; **, p < 0.01; ***, p < 0.001.