MOESM1 of Deficiency of biodegradable plastic-degrading enzyme production in a gene-deletion mutant of phyllosphere yeast, Pseudozyma antarctica defective in mannosylerythritol lipid biosynthesis
2019-07-06T05:00:00Z (GMT) by
Additional file 1: Figure S1. PCR analysis of the parental strain GB-4(0) and strain ΔPaEMT1. The primer sets used in PCR amplification to assess PaEMT1 deletion (A). Agarose gel electrophoresis of amplified DNA fragments to confirm gene disruption. The gene fragments were amplified using PaEMT1_inner_F1 (Primer A) and PaEMT1_inner_R1 (Primer B), or PaEMT1_up_F2 (Primer C) and NATfragment_R1 (Primer D) (B). Figure S2. Schematic diagram of the procedure used to obtain cell-free extract. Figure S3. PaE production by strain ΔPaEMT1 in m-3×FMM medium supplemented with 8 % xylose and various concentrations of TritonX-100. Cell growth (white) and PaE activity (gray) (A) and SDS-PAGE and western blot analyses of culture supernatant (B). CMC, critical micelle concentration; M, marker; P, purified PaE. The amount of each culture supernatant loaded in the gel was 10 μl for CBB staining and western blotting. The results of the cell growth and PaE activity assays are shown as the average of three different experiments. Error bars show standard deviations. Figure S4. TLC analysis of culture supernatants used for the PaE activity assay. Culture supernatants (A) and precipitates including cells (B).