Figure legends
Figure 1. Interaction analysis of DEGs and DEPs. (A)Correction plot analysis based on DEGs and DEPs. (B) Venn diagram of DEGs and DEPs. (C) Heat map based on FPKM value of DEGs and DEPs.
Figure 2 . Diagram of the secondary and tertiary structure of the mutant gene. (A) Comparison of amino acid sequences of alpha-amylases and CYP6AE76. Homologous sequence regions 1, 2, 3 and 4 are surrounded by purple rectangles. The amino acid sequence described above the rectangular regions was taken as representative of regions 1 to 4, respectively. Active sites and those of substrate binding proposed by Matsuura et al. (Matsumura et al., 1984) for Taka-amylase A from are indicated by the purple triangle and black oval, respectively; SRS is represented in green line boxes, heme-binding signature motif (FxxGxxxCxG) (black-dotted rectangular frame), helix C motif (WxxxR) (navy blue box), and PxPF motif are in lightboxes.(B) superimpositions of predicted models of α-amylase and CYP6AE76 from C. pinicolalis and C. punctiferalis with their respective templates.
Figure 3. Gene expression of α-amylase and P450 monooxygenase CYP6AE76 in C. pinicolalisand C. punctiferalis . (A) The relative expression of α-amylase in C. pinicolalis and C. punctiferalis .(B) The relative expression of P450 monooxygenase CYP6AE76 inC. pinicolalis and C. punctiferalis . The gene expression level between the two species was statistically significant (t -test, ***P < 0.001).
Figure 4. The comparison of enzymatic activity of recombinant α-amylase and CYP6AE76 from two species. (A) The amount of ethylidene-pNP-G7 (substrate) cleaved by the purified α-amylase from C. pinicolalis and C. punctiferalis . (B) Conversion of p-nitroanisole to p-nitrophenol in by recombinant cytochrome P450 from C. pinicolalis and C. punctiferalis . The α-amylase and CYP6AE76 enzyme activities were statistically significant (t -test,**P <0.01).
Figure 5. Association analysis the different genes between the two species (C. pinicolalis vs C. punctiferalis ).(A) Volcano map of all differential genes;(B) Top 20 down- and up-accumulated metabolites between the two species.
Figure S1. Transcriptome and proteomics association analysis.(A) Differential gene GO enrichment. (B) and(C) Differential gene GO enrichment of biological process and molecular function, respectively.
Figure S2. KEGG pathway enrichment scatter plot of the different genes.
Figure S3. Candidate gene recombinant expression protein.(A) SDS-PAGE analysis of purified recombinant Candidate proteins. The four SDS-PAGE diagrams are Coomassie Blue staining of purified α-amylase (1 and 2) and CYP6AE76 (3 and 4) protein samples from left to right in C. pinicolalis vs C. punctiferalis , respectively. M: Protein marker; Lane 1,2,5,6,9,10,13 and 14: Sonicated bacterial pellet; 3,4,7,8,11,12,15 and 16: purified protein.(B) Western blot analysis of α-amylase (Line 1 and 2) and CYP6AE76 (Line 3 and 4) protein samples from left to right in C. pinicolalis vs C. punctiferalis , respectively. M: Protein marker.
Figure S4. Metabolite KEGG classification map.
Figure S5. KEGG pathway enrichment scatter plot of the different metabolite.