Phylogenetic analysis of β-xylanase SRXL1 of Sporisorium reilianum and its relationship with families (GH10 and GH11) of Ascomycetes and Basidiomycetes

Abstract

In this paper, the amino acid sequence of the β-xylanase SRXL1 of Sporisorium reilianum, which is a pathogenic fungus of maize was used as a model protein to find its phylogenetic relationship with other xylanases of Ascomycetes and Basidiomycetes and the information obtained allowed to establish a hypothesis of monophyly and of biological role. 84 amino acid sequences of β-xylanase obtained from the GenBank database was used. Groupings analysis of higher-level in the Pfam database allowed to determine that the proteins under study were classified into the GH10 and GH11 families, based on the regions of highly conserved amino acids, 233-318 and 180-193 respectively, where glutamate residues are responsible for the catalysis.

Figure 1

Alignment of amino acid sequence of the active site region of the family GH11 xylanases: M. oryzae (G4NA54, G4MVY2, G4N696, G4MWS3), M. grisea (Q92245, P0CT48, Q92244), F. oxysporum (F9F5R3, Q9C1R1, Q9C1R2, F9FP27, F9FI26), F. graminearum (Q7ZA57), C. carbonum (Q06562, Q00350), T. lanuginosus (F8UV78, O43097), T. reesei (p36217, P36218), T. harzianum (P48793), A. niger (P55330, P55329, Q12550, Q6QJ75, Q12549), E. nidulans (P55333, P55332), A. pullulans (Q9UW17, Q12562, Q96TR7), S. sclerotiorum (A7EXM7, A7EQZ6), P. variotii (P81536), C. cinerea (A8P8F0), P. chrysosporium (Q9HEZ0, Q9HEY9), U. maydis (Q4P0L3), P. ostreatus (B0FX60). And family GH10: M. oryzae (G4MLU0, G4NIM7, G4NBN8), M. grisea (Q8JIY4, Q01176), F. oxysporum (B3A0S5, O59938), C. carbonum (Q6GXE5), A. niger (G3Y866, C5J411), A. pullulans (Q2PGV8), T. virens (G9NBD2), P. purpurogenum (Q9P8J1), P. oxalicum (E1B2N4, HQ157197), T. aerugineus (G8ZAH1), T. aurantiacus (P23360), A. kawachii (P33559, GAA92552, JT0608), A. oryzae (O94163, Q96VB6), A. fumigatus (Q0H904), A. versicolor (A2I7V1), A. sojae (Q9P955, BAA92882), A. terreus (Q4JHP5, AFD63136, H9BYX9, Q0CBM8), A. aculeatus (O59859), P. crysogenum (B6F253), P. citrinum (BAG12101, B1B533), P. simplicissimum (P56588), P. canescens (Q5S7A8), C. cinerea (A8P570), U. hordei (I2FWP8), S. reilianum (E7A3D3), S. commune (D8Q1J8), T. versicolor (EIW54190), P. strigosozonata (EIN11616), P. chrysosporium (Q9HEZ2, Q9HEZ1, AAG44993, G0ZCU2). Gaps are indicated by a hyphen (–). A standard numbering system for the active site is included up of each block of sequences. The motives conserved in xylanases are indicated in blue that corresponds to the glutamates of the active site.

Figure 2

Phylogenetic analysis of the amino acid sequences of xylanases from Ascomycetes (blue letters) and Basidiomycetes (red letters) of the families GH10 (Access codes on green) and GH11 (Access codes on orange). The numbers located in the clades indicate the estimated time of divergence for enzymes in millions of years (Myr).

Figure 3

Motifs elements present in the fungal xylanases of the family GH10 (A): P. chrysosporyum, Q9HEZ1; A. sojae, Q9P955; M. grisea, Q8J1Y4; S. reilianum, E7A3D3; U. hordei, I2FWP8; A. versicolor, A2I7V1; S. commune, 8Q1J8; T. virens, G9NBD2; T. versicolor, EIW54190; C. carbonum, Q6GXE5; F. oxysporum, O59938; M. grisea, Q01176 y GH11 (B): F. oxysporum, Q9C1R1; M. grisea, Q92245; M. oryzae, G4NA54; A. niger, Q6QJ75; A. pullulans, Q96TR7; F. graminearum, Q7ZA57; A. niger, P55330; E. nidulans, P55333; P. ostreatus, B0FX60; C. cinerea, A8P8F0; C. carbonum, Q00350; C. carbonum, Q06562; E. nidulans, P55332; U. maydis, Q4P0L3; M. grisea, P0CT48. The numbers indicate the number of amino acid or position of signal peptide (SP). The region conserved in all proteins is shown in blue. Glutamates responsible for catalysis are indicated by the letter E followed by a number that shows its position in the sequence.

Figure 4

Relationship of the theoretical structure of the xylanases of GH10 family (A) P. chrysosporyum, Q9HEZ1; A. sojae, Q9P955; M. grisea, Q8J1Y4; S. reilianum, E7A3D3; U. hordei, I2FWP8; A. versicolor, A2I7V1; S. commune, 8Q1J8; , G9NBD2; T. versicolor, EIW54190; C. carbonum, Q6GXE5; F. oxysporum, O59938; M. grisea, Q01176 and GH11 (B) F. oxysporum, Q9C1R1; M. grisea, Q92245; M. oryzae, G4NA54; A. niger, Q6QJ75; A. pullulans, Q96TR7; F. graminearum, Q7ZA57; A. niger, P55330; E. nidulans, P55333; P. ostreatus, B0FX60; C. cinerea, A8P8F0; C. carbonum, Q00350; C. carbonum, Q06562; E. nidulans, P55332; U. maydis, Q4P0L3; M. grisea, P0CT48. With respect to time of appearance (Myr). The modeling was carried as described in methodology.