Supporting Information

Analyzing the Substrate Specificity of a Class of Long-Horned--Derived Xylanases by Using Synthetic Arabinoxylan Oligo- and Polysaccharides Yannick Pauchet,*[a] Colin Ruprecht,[b] and Fabian Pfrengle[b, c]

cbic_201900687_sm_miscellaneous_information.pdf Table S1. Details on the Cerambycidae-derived GH5_2 sequences used for the phylogenetic analysis presented in Figure 2 and for the amino acid alignment presented in Figure S1.

Acronym Species Family Subfamily Function Reference AJA1 Apriona japonica Cerambycidae Xylanase (1) AJA2 Apriona japonica Cerambycidae Lamiinae Cellulase (1) AJA3 Apriona japonica Cerambycidae Lamiinae Cellulase (1) AJA4 Apriona japonica Cerambycidae Lamiinae Cellulase (1) AJA5 Apriona japonica Cerambycidae Lamiinae No activity (1) AJA6 Apriona japonica Cerambycidae Lamiinae Cellulase (1) AGL1 Anoplophora glabripennis Cerambycidae Lamiinae Xylanase (2) AGL2 Anoplophora glabripennis Cerambycidae Lamiinae Cellulase (2) AGL3 Anoplophora glabripennis Cerambycidae Lamiinae No activity (2) AGL4 Anoplophora glabripennis Cerambycidae Lamiinae No activity (2) AGL5 Anoplophora glabripennis Cerambycidae Lamiinae Cellulase (2) AGL6 Anoplophora glabripennis Cerambycidae Lamiinae Cellulase (2) MAL1 Monochamus alternatus Cerambycidae Lamiinae Not tested a MAL2 Monochamus alternatus Cerambycidae Lamiinae Not tested a MAL3 Monochamus alternatus Cerambycidae Lamiinae Not tested a MAL4 Monochamus alternatus Cerambycidae Lamiinae Not tested a MAL5 Monochamus alternatus Cerambycidae Lamiinae Not tested a MAL6 Monochamus alternatus Cerambycidae Lamiinae Not tested a MMYA myops Cerambycidae Lamiinae Not tested (3) MMYB Cerambycidae Lamiinae Not tested (3) MMYC Mesosa myops Cerambycidae Lamiinae Not tested (3) MMYD Mesosa myops Cerambycidae Lamiinae Not tested (3) MMYE Mesosa myops Cerambycidae Lamiinae Not tested (3) MMYF Mesosa myops Cerambycidae Lamiinae Not tested (3) OAL Oncideres albomarginata chamela Cerambycidae Lamiinae Not tested (4) PHI Cerambycidae Lamiinae Cellulase (5) AGE Cerambycidae Lamiinae Cellulase (6) ACH Anoplophora chinensis Cerambycidae Lamiinae Cellulase (7) aGH5_2 sequences were curated from an assembly of RNA-Seq data (SRA accessions: SRX1302202; SRX1605798; SRX1605842; SRX1606425; SRX1606426). 1. Pauchet Y, Kirsch R, Giraud S, Vogel H, Heckel DG. Identification and characterization of plant cell wall degrading enzymes from three glycoside hydrolase families in the cerambycid beetle Apriona japonica. Biochem Mol Biol. 2014;49:1-13. 2. McKenna DD, Scully ED, Pauchet Y, Hoover K, Kirsch R, Geib SM, et al. Genome of the Asian longhorned beetle (Anoplophora glabripennis), a globally significant invasive species, reveals key functional and evolutionary innovations at the beetle-plant interface. Genome biology. 2016;17(1):227. 3. Liu J, Song K, Teng H, Zhang B, Li W, Xue H, et al. Endogenous cellulolytic enzyme systems in the Mesosa myops (Insecta: Coleoptera) studied by transcriptomic analysis. Acta biochimica et biophysica Sinica. 2015;47(9):741-8. 4. Calderon-Cortes N, Watanabe H, Cano-Camacho H, Zavala-Paramo G, Quesada M. cDNA cloning, homology modelling and evolutionary insights into novel endogenous cellulases of the borer beetle Oncideres albomarginata chamela (Cerambycidae). Insect molecular biology. 2010;19(3):323-36. 5. Sugimura M, Watanabe H, Lo N, Saito H. Purification, characterization, cDNA cloning and nucleotide sequencing of a cellulase from the yellow-spotted longicorn beetle, Psacothea hilaris. European Journal of Biochemistry. 2003;270(16):3455-60. 6. Wei YD, Lee KS, Gui ZZ, Yoon HJ, Kim I, Zhang GZ, et al. Molecular cloning, expression, and enzymatic activity of a novel endogenous cellulase from the mulberry longicorn beetle, Apriona germari. Comparative biochemistry and physiology Part B, Biochemistry & molecular biology. 2006;145(2):220-9. 7. Chang CJ, Wu CP, Lu SC, Chao AL, Ho TH, Yu SM, et al. A novel exo-cellulase from white spotted longhorn beetle (Anoplophora malasiaca). Insect Biochem Mol Biol. 2012;42(9):629-36.

Figure S1. Amino acid alignment of cerambycid-derived GH5_2 proteins. The predicted signal peptide was removed from each protein sequence prior to alignment. The degree of amino acid conservation is indicated by purple shadings. A GH5_2 sequence derived from the bacterium Zobellia galactanivorans (ZGA, CAZ97841.1) was used as an outgroup. Details on the sequences used here can be found in Table 1. The two catalytic glutamic acid residues are indicated by red arrows. Residues implicated in binding are indicated by black arrows. AJA: Apriona japonica; AGL: Anoplophora glabripennis; AGE: Apriona germari; PHI: Psacothea hilaris; OAL: Oncideres albomarginata chamela; ACH: Anoplophora chinensis; MMY: Mesosa myops; MAL: Monochamus alternatus.