Fig. 4

Identification of lipase protein and gene sequences in ZS6. a In-gel lipase assay. Left panel shows the native PAGE analysis of proteins prepared from supernatant of ZS6 culture. The right panel shows the in-gel analysis of lipases. Bright spots indicate the increase of acidity caused by lipid hydrolysis. b Lipase shared with trypsinized peptide sequences generated in LC–MS/MS analysis. Residue at the end of trypsin peptides were highlighted in bold. Peptide sequences identified by MS analysis are highlighted. Residue at the end of trypsin peptides were highlighted in bold. c Probable lipase protein in SDS-PAGE gel. d Probable lipase encoding DNA is amplified using PCR analysis. Amino acid substitutes between E13 Lip and ZS6 Lip are shown in the bottom panel. Five amino acid substitutions were found in the deduced protein sequences compared with that of the E13 LipA. e Sequence-based lipase phylogenetic tree analysis. Sequences whose Genbank accession number (ZS6, MG897498.1) followed by the lipase family [I–VIII families (Arpigny and Jaeger 1999)] is shown. g Levels of lipase gene expression at 8 h and 16 h after growth based on RT-qPCR analysis are shown