title: Thermostable beta-glycosidase-CBD fusion protein for biochemical analysis of cotton scouring efficiency.
abstract: Multidomain proteins for the biochemical analysis of the scouring efficiency of cotton fabrics were constructed by the fusion of a reporter moiety in the N-terminal and the cellulose binding domain (CBD) in the C-terminal. Based on the specific binding of the CBD of Cellulomonas fimi exoglucanase (Cex) to crystalline cellulose (Avicel), the reporter protein is guided to the cellulose fibers that are increasingly exposed as the scouring process proceeds. Among the tested reporter proteins, a thermostable beta-glycosidase (BglA) from Thermus caldophilus was found to be most appropriate, showing a higher applicability and stability than GFP, DsRed, or a tetrameric beta-glucuronidase (GUS) from Escherichia coli, which were precipitated more seriously during the expression and purification steps. When cotton fabrics with different scouring levels were treated with the BglA-CBD and incubated with X-Gal as the chromogenic substrate, an indigo color became visible within 2 h, and the color depth changed according to the conditions and extent of the scouring.
authors.affiliation: Systems Microbiology Research Center, KRIBB, Daejeon 305-806, Korea.
authors.forename: Jae-Seok
authors.initials: JS
authors.lastname: Ha
authors.forename: Young-Mi
authors.initials: YM
authors.lastname: Lee
authors.forename: Su-Lim
authors.initials: SL
authors.lastname: Choi
authors.forename: Jae Jun
authors.initials: JJ
authors.lastname: Song
authors.forename: Cheol-Soo
authors.initials: CS
authors.lastname: Shin
authors.forename: Ju-Hea
authors.initials: JH
authors.lastname: Kim
authors.forename: Seung-Goo
authors.initials: SG
authors.lastname: Lee
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D044602:Cellulases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D040143:Cellulomonas / Q000201:enzymology / Q000235:genetics; D002482:Cellulose / Q000032:analysis / Q000378:metabolism*; D032561:Cotton Fiber; D007203:Indigo Carmine; D007211:Indoles / Q000032:analysis; D008969:Molecular Sequence Data; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D013824:Thermus / Q000201:enzymology / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D007211:Indoles; D011993:Recombinant Fusion Proteins; D002482:Cellulose; D007203:Indigo Carmine; D044602:Cellulases