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oai:figshare.com:article/31414090 2026-09-17T19:09:14Z category_1 category_4 category_7 category_8 category_12 category_21 category_873 category_135 portal_63 item_type_2 month_year_09_2026

Reporter Group-Labeled Synthetic Cellulose: Structural Characterization and Utilization in Mapping the Cellulose Chain-Cleavage Modes of Cellulases Gaurav Singh Kaira (23254921) Manuel Eibinger (1452625) Chao Zhong (2247550) Bernd Nidetzky (86451) Biophysics Biochemistry Medicine Microbiology Cell Biology Biotechnology Chemical Sciences not elsewhere classified Computational Biology residual solid material pnp )- β local nanoscale structure crystalline solid substrate clostridium thermocellum cellulosome cellulases iterative β trichoderma reesei cellulases processive chain cleavages chain cleavage specificities 4 – 10 end reporter group labeled synthetic cellulose reporter group synthetic cellulose reesei </ cleavage modes chain ends cellulose chain labeled cellulose labeled cellooligosaccharides ∼ 5 upon self synthesize reducing structural characterization soluble products show synthesis products formed oligosaccharide composition mass spectrometry like assemblies hydrolase systems directionality dependent degrading enzymes degradation efficiency cellodextrin phosphorylase average degree Iterative β-1,4-glucosylation of p-nitrophenyl (pNP)-β-cellobioside by cellodextrin phosphorylase was applied to synthesize reducing-end reporter group-labeled cellooligosaccharides (average degree of polymerization, DP: ∼5.7; DP range: 4–10) which, upon self-assembly in solution, precipitate as a cellulose allomorph II crystalline material. Atomic force microscopy revealed that the synthetic cellulose-pNP forms nanoscale sheet-like assemblies, with thickness ∼4 nm. The labeled cellulose was used to characterize the chain cleavage specificities of cellulose-degrading enzymes. Reactions were monitored by the release of soluble products as well as the change in oligosaccharide composition of the residual solid material, measured by mass spectrometry. The products formed by hydrolase systems (Trichoderma reesei cellulases; Clostridium thermocellum cellulosome) are controlled by processive chain cleavages from the chain ends. Atomic force microscopy shows that <i>T. reesei</i> Cel7A preferentially attacked sheet edges, with degradation efficiency and directionality dependent on local nanoscale structure. Collectively, we show synthesis of reporter group-labeled cellulose for probing enzymatic chain depolymerization activity on a crystalline solid substrate. 2026-03-09T00:00:00Z Dataset Media 10.1021/acs.biomac.5c02515.s002 https://figshare.com/articles/media/Reporter_Group-Labeled_Synthetic_Cellulose_Structural_Characterization_and_Utilization_in_Mapping_the_Cellulose_Chain-Cleavage_Modes_of_Cellulases/31414090 CC BY-NC 4.0