β-Glucosidase (BGL) is a key rate-limiting enzyme in the cellulose degradation system.To elucidate the molecular features and functional mechanisms of Trichoderma BGLs, 137 validated sequences were retrieved from the NCBI database.The analyses included sequence classification, physicochemical properties, structural features, and phylogenetic relationships.Conserved motifs, domains, tertiary structure modeling, molecular docking, and protein-protein interaction networks were also examined.The results showed that Trichoderma BGLs were distributed across 25 species and belong to four families, including GH1, GH3, GH17, and GH132, with GH3 being predominant.GH3 and GH1 proteins were enriched in α-helices and β-sheets, displaying high stability and hydrophilicity, whereas GH132 and GH17 contained abundant random coils, showing lower stability but greater flexibility.Subcellular localization indicated that 56.2% (77) were secreted extracellularly.Phosphorylation prediction indicated that BGLs were mainly regulated through reversible serine/threonine phosphorylation.GH3 BGLs from Trichoderma reesei contained conserved motifs and a BglX domain, and they cooperated with cell wall-degrading enzymes and carbohydrate metabolism proteins.Molecular docking demonstrated that the blue pocket of the representative GH3 member GFP60323.1 bound cellobiose most stably, while the orange pocket accommodated diverse aromatic substrates.In conclusion, this study systematically characterized the structural and functional properties of Trichoderma BGLs and provided new insights into their potential applications in bioenergy development, food processing, and feed industry.
[1] ZHAN Q W, LIN Q X, WU Y, et al.A fractionation strategy of cellulose, hemicellulose, and lignin from wheat straw via the biphasic pretreatment for biomass valorization[J].Bioresource Technology, 2023, 376:128887.
[2] HOUFANI A A, ANDERS N, SPIESS A C, et al.Insights from enzymatic degradation of cellulose and hemicellulose to fermentable sugars-a review[J].Biomass and Bioenergy, 2020, 134:105481.
[3] BANU J R, SUGITHA S, KAVITHA S, et al.Lignocellulosic biomass pretreatment for enhanced bioenergy recovery:Effect of lignocelluloses recalcitrance and enhancement strategies[J].Frontiers in Energy Research, 2021, 9:646057.
[4] SENGUPTA S, DATTA M, DATTA S.Glycoside Hydrolases[M].Amsterdam:Elsevier, 2023:97-120.
[5] YANG W Q, SU Y W, WANG R B, et al.Microbial production and applications of β-glucosidase-A review[J].International Journal of Biological Macromolecules, 2024, 256:127915.
[6] SHARMA J, KUMAR V, PRASAD R, et al.Engineering of Saccharomyces cerevisiae as a consolidated bioprocessing host to produce cellulosic ethanol:Recent advancements and current challenges[J].Biotechnology Advances, 2022, 56:107925.
[7] GUTIERREZ-GUTIERREZ D A, FUENTES-GARIBAY J A, VIADER-SALVADÓ J M, et al.Biochemical characterization of the β-glucosidase Glu1B from Coptotermes formosanus produced in Pichia pastoris[J].Enzyme and Microbial Technology, 2023, 163:110155.
[8] KANNAN P, SHAFREEN M M, ACHUDHAN A B, et al.A review on applications of β-glucosidase in food, brewery, pharmaceutical and cosmetic industries[J].Carbohydrate Research, 2023, 530:108855.
[9] ZHU Q, HUANG Y Y, YANG Z F, et al.A recombinant thermophilic and glucose-tolerant GH1 β-glucosidase derived from Hehua hot spring[J].Molecules, 2024, 29(5):1017.
[10] 王可心. β-葡萄糖苷酶多种酶学特性的协同进化及其机制[D].南昌:江西师范大学, 2022.
WANG K X.Synergetic evolution and its mechanism of multiple enzymatic properties of β-glucosidase[D].Nanchang:Jiangxi Normal University, 2022.
[11] GODSE R, BAWANE H, TRIPATHI J, et al.Unconventional β-glucosidases:A promising biocatalyst for industrial biotechnology[J].Applied Biochemistry and Biotechnology, 2021, 193(9):2993-3016.
[12] TIWARI P, MISRA B N, SANGWAN N S.β-glucosidases from the fungus Trichoderma:An efficient cellulase machinery in biotechnological applications[J].BioMed Research International, 2013, 2013(1):203735.
[13] LIMA P C, KARIMIAN P, JOHNSTON E, et al.The use of Trichoderma spp.for the bioconversion of agro-industrial waste biomass via fermentation:A review[J].Fermentation, 2024, 10(9):442.
[14] MOU L, PAN R Z, LIU Y S, et al.Isolation of a newly Trichoderma asperellum LYS1 with abundant cellulase-hemicellulase enzyme cocktail for lignocellulosic biomass degradation[J].Enzyme and Microbial Technology, 2023, 171:110318.
[15] TAMURA K, STECHER G, KUMAR S.MEGA11:Molecular evolutionary genetics analysis version 11[J].Molecular Biology and Evolution, 2021, 38(7):3022-3027.
[16] SERRA G M, SANTOS SIQUEIRA A, DE MOLFETTA F A, et al.In silico analysis of a GH3 β-glucosidase from Microcystis aeruginosa CACIAM 03[J].Microorganisms, 2023, 11(4):998.
[17] MOHAMAD SOBRI M F, ABD-AZIZ S, ABU BAKAR F D, et al.In-silico characterization of glycosyl hydrolase family 1 β-glucosidase from Trichoderma asperellum UPM1[J].International Journal of Molecular Sciences, 2020, 21(11):4035.
[18] LIANG Y P, UNIVERISITY Y, ZHAO Y L, et al.Conserved local structural motifs in glycoside hydrolase families facilitate the discovery of functional enzymes[J].Journal of Agricultural and Food Chemistry, 2025, 73(19):11983-11997.
[19] DOUGHERTY D A.The cation- π interaction in chemistry and biology[J].Chemical Reviews, 2025, 125(5):2793-2808.
[20] SIDAR A, VOSHOL G P, ARENTSHORST M, et al.Deciphering domain structures of Aspergillus and Streptomyces GH3-β-Glucosidases:A screening system for enzyme engineering and biotechnological applications[J].BMC Research Notes, 2024, 17:257.
[21] HRMOVA M, ZIMMER J, BULONE V, et al.Enzymes in 3D:Synthesis, remodelling, and hydrolysis of cell wall (1,3;1,4)-β-glucans[J].Plant Physiology, 2023, 194(1):33-50.
[22] ZHANG X J, DUAN P L, SHI S Q, et al.Global characterization of GH3 family glycoside hydrolase genes in Fusarium verticillioides and functional analysis of FvGH3-6[J].Frontiers in Microbiology, 2025, 16:1543210.
[23] YANG Y Z, WANG J, GUO H N, et al.The enzymatic characters of heterologous expressed novel β-1,4-glucosidase originated from Aspergillus fresenii[J].3 Biotech, 2020, 10(6):239.
[24] PANG A P, WANG H Y, LUO Y S, et al.Dissecting cellular function and distribution of β-glucosidases in Trichoderma reesei[J].mBio, 2021, 12(3):e03671-e03620.
[25] ROY A, KALITA B, THANMALAGAN R R, et al.Differential expression and gene correlation analyses reveal core CAZymes in Fusarium oxysporum f.sp.lycopersici exposed to mutagen and heat stress[J].Journal of Phytopathology, 2024, 172(1):e13254.
[26] LINARES-PASTÉN J A, JONSDOTTIR L B, HREGGVIDSSON G O, et al.Modeled 3D-structures of proteobacterial transglycosylases from glycoside hydrolase family 17 give insight in ligand interactions explaining differences in transglycosylation products[J].Applied Sciences, 2021, 11(9):4048.
[27] DEFLANDRE B, JADOT C, PLANCKAERT S, et al.Structure and function of BcpE2, the most promiscuous GH3-family glucose scavenging beta-glucosidase[J].mBio, 2022, 13(4):e00935-e00922.
[28] ZHANG J, ZHAO N, XU J N, et al.Homology analysis of 35 β-glucosidases in Oenococcus oeni and biochemical characterization of a novel β-glucosidase BGL0224[J].Food Chemistry, 2021, 334:127593.
[29] TEUFEL F, ALMAGRO ARMENTEROS J J, JOHANSEN A R, et al.SignalP 6.0 predicts all five types of signal peptides using protein language models[J].Nature Biotechnology, 2022, 40(7):1023-1025.
[30] HALLGREN J, TSIRIGOS K D, PEDERSEN M D, et al.DeepTMHMM predicts alpha and beta transmembrane proteins using deep neural networks[J].bioRxiv, 2022.https://doi.org/10.1101/2022.04.08.487609.
[31] DE ASSIS M A, DA SILVA J J B, DE CARVALHO L M, et al.A multiomics perspective on plant cell wall-degrading enzyme production:Insights from the unexploited fungus Trichoderma erinaceum[J].Journal of Fungi, 2024, 10(6):407.
[32] WANG H, LIN X N, LI S, et al.Rational molecular design for improving digestive enzyme resistance of beta-glucosidase from Trichoderma viride based on inhibition of bound state formation[J].Enzyme and Microbial Technology, 2020, 133:109465.
[33] HU C X, WANG Y B, WANG W Y, et al.A trapped covalent intermediate as a key catalytic element in the hydrolysis of a GH3 β-glucosidase:An X-ray crystallographic and biochemical study[J].International Journal of Biological Macromolecules, 2024, 265:131131.
[34] SIVARAMAKRISHNAN M, VEERAGANTI NAVEEN PRAKASH C, CHANDRASEKAR B.Multifaceted roles of plant glycosyl hydrolases during pathogen infections:More to discover[J].Planta, 2024, 259(5):113.
