title: Cellulose hydrolysis ability of a Clostridium thermocellum cellulosome containing small-size scaffolding protein CipA.
abstract: Mutant Clostridium thermocellum YM72 that produces small-size scaffolding protein CipA (ssCipA) was isolated from wild-type YM4. Sequencing of ssCipA revealed that two domains, cohesin 6 and cohesin 7, were not present. Cellulosome prepared from YM72 exhibited a significant reduction of hydrolysis ability on crystalline celluloses such as Sigmacell type-20 and cellulose from Halocynthia. To investigate this influence in vitro, artificial cellulosomes were assembled as recombinant CipA (rCipA) and ssCipA (rssCipA) using native free-cellulosomal subunits. The cellulosome assembled using rssCipA showed a 1.8-fold decrease in the hydrolysis of crystalline cellulose compared with that of rCipA. However, no significant differences in the hydrolysis of carboxymethylcellulose and acid-swollen cellulose were observed. One protein band was missing from the complex that was assembled using rssCipA (confirmed by native-PAGE). The missing protein was identified as CelJ, which is a major cellulosomal subunit. This suggests that insufficient cooperation of CelJ into the cellulosome results in the significant reduction of hydrolysis toward crystalline cellulose. These results indicate that cohesin 6 and 7 may be responsible for the cooperation of CelJ through cohesin and dockerin interactions, and adequate cooperation of CelJ into the cellulosome is important for significant hydrolysis of crystalline cellulose.
authors: Biological Resources and Post-harvest Division, Japan International Research Center for Agricultural Sciences (JIRCAS), 1-1 Ohwashi, Tsukuba, Ibaraki 303-8686, Japan.
authors: Lan
authors: L
authors: Deng
authors: Biological Resources and Post-harvest Division, Japan International Research Center for Agricultural Sciences (JIRCAS), 1-1 Ohwashi, Tsukuba, Ibaraki 303-8686, Japan.
authors: Yutaka
authors: Y
authors: Mori
authors: Biological Resources and Post-harvest Division, Japan International Research Center for Agricultural Sciences (JIRCAS), 1-1 Ohwashi, Tsukuba, Ibaraki 303-8686, Japan.
authors: Junjarus
authors: J
authors: Sermsathanaswadi
authors: University of Tsukuba Graduate School of Life and Environmental Sciences, 1-1-1 Tennoudai, Tsukuba, Ibaraki 305-8572, Japan.
authors: Waraporn
authors: W
authors: Apiwatanapiwat
authors: Biological Resources and Post-harvest Division, Japan International Research Center for Agricultural Sciences (JIRCAS), 1-1 Ohwashi, Tsukuba, Ibaraki 303-8686, Japan; University of Tsukuba Graduate School of Life and Environmental Sciences, 1-1-1 Tennoudai, Tsukuba, Ibaraki 305-8572, Japan. Electronic address: akosugi@affrc.go.jp.
authors: Akihiko
authors: A
authors: Kosugi
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000235:genetics; D002480:Cellulase / Q000235:genetics / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism*; D044603:Cellulosomes / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics; D048013:Acetivibrio thermocellus / Q000235:genetics / Q000378:metabolism*; D006868:Hydrolysis; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D000097722:Cohesins
chemical_list: D001426:Bacterial Proteins; D018797:Cell Cycle Proteins; D002480:Cellulase; D002482:Cellulose; D002868:Chromosomal Proteins, Non-Histone; D008565:Membrane Proteins; D011994:Recombinant Proteins; D000097722:Cohesins; C080369:CipA protein, Clostridium; C101903:CelJ protein, Clostridium thermocellum
keywords: CelJ; Cellulosome; Clostridium thermocellum; Cohesin; Crystalline cellulose