Fig. 2.

Fig. 2. Protein domains and their phylogenetic relationships with typical bacterial collagenases or collagenolytic proteases. (A) Schematic representations of the maturase domain organization of typical collagenases or collagenolytic proteases from different microorganisms. All the domain structures were deduced from the amino acid sequences of TSS (OBR56241) from Brevibacillus sp. WF146 [17], SOT (BAI44325) from Streptomyces omiyaensis [46], kumamolisin-As (BAC41257) from Alicyclobacillus sendaiensis [53], MO-1 (AB260948) from Geobacillus collagenovorans MO-1 [93], MCP-01 (ABD14413) from Pseudoalteromonas sp. SM9913 [67], myroicolsin (AEC33275) from Myroides profundi D25 [65], AcpII (AB505451) from Alkalimonas collagenimarina [83], VMC (AAC23708) from Vibrio mimicus [83], VHC (BAK39964) from Grimontia (Vibrio) hollisae [94] and ColG (BAA77453) from Clostridium histolyticum [11]. CD: catalytic domain [19]; βJR: β-jelly roll domain [17, 21]; CBD: collagen-binding domain [19]; P: P-proprotein convertase domain [20]; PKD: polycystic kidney disease-like domain [20]; PPC: pre-peptidase C-terminal domain [20]; AD: activator domain [91]; PA: protease-associated domain [92]. (B) A rootless phylogenetic tree was constructed from the amino acid sequence alignment of full-length enzymes using the neighbor-joining method in ClustalX and MEGA7 to investigate the evolutionary relationship between TSS and other S8 family subtilases. For the proteases displayed above, the enzymes were divided into the following groups: true subtilisins; HAPs (high-alkaline proteases); ICPs (intercellular proteases); OSPs (oxidatively stable proteases); HMPs (high-molecular-mass proteases); PISs (phylogenetically intermediate subtilisins); thermitase; proteinase K; pyrolysin; and Kexin and lantibiotic peptidase. The origins of the sequences aligned: Kexin (OLN81751) from Colletotrichum chlorophyti; furin isoform X1 (XP_011249120) from Mus musculus (house mouse); Vpr (M76590) from Bacillus subtilis; Bha (G83753) from Bacillus halodurans C-125; lantibiotic (KJS88019) from Desulfosporosinus sp. BICA1–9; KP-43 (AB051423) from Bacillus sp. strain KSM-KP43; KP-9860 (AB046403) from Bacillus sp. strain KSM-KP9860; INT72 (P29139) from Bacillus polymyxa 72; Isp-Q (Q45621) from Bacillus sp. strain NKS-21; pyrolysin (AAB09761) from Pyrococcus furiosus DSM 3638; proteinase K (1205229A) from Parengyodontium album; PR1A (AAV97788) from Metarhizium acridum; thermitase (KAA1806649) from Bacillus cereus; subtilisin Carlsberg (2SEC_E) from Bacillus licheniformis; BPN’ (Q44684) from Bacillus amyloliquefaciens; subtilisin E (P04189) from Bacillus subtilis 168; LD1 (AB085752) from Bacillus sp. strain KSM-LD1; ALP-1 (Q45523) from Bacillus sp. strain NKS-21; M-protease (Q99405) from Bacillus clausii KSM-K16; MO-1 (AB260948) from Geobacillus sp. MO- 1; myroicolsin (AEC33275) from Myroides profundi; MCP-01 (ABD14413) from Pseudoalteromonas sp. SM9913; AcpII (AB505451) from Alkalimonas Collagenimarina; and TSS (1039472844) from Brevibacillus sp. WF146. Collagenolytic proteases from S8 subtilases, including myroicolsin, MCP-01, AcpII, and MO-1, are represented as green dots, whereas TSS is represented using an orange asterisk. (C) Homology modeling and structural fitting chart of TSS (RoseTTAFold, https://github.com/RosettaCommons/RoseTTAFold) and KP-43 (PDB code1WMF) by SpdbViewer. The catalytic and βJR domains were represented by light blue and green for TSS, and purple and gray for KP-43, respectively. The side chains of the catalytic triad of TSS (D-H-S) were shown in black.

© J. Microbiol. Biotechnol.