Thermostable beta-glycosidase-CBD fusion protein for biochemical analysis of cotton scouring efficiency.
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.
Ha
Jae-Seok
JS
Systems Microbiology Research Center, KRIBB, Daejeon 305-806, Korea.
Lee
Young-Mi
YM
Choi
Su-Lim
SL
Song
Jae Jun
JJ
Shin
Cheol-Soo
CS
Kim
Ju-Hea
JH
Lee
Seung-Goo
SG
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
D001426:Bacterial Proteins; D007211:Indoles; D011993:Recombinant Fusion Proteins; D002482:Cellulose; D007203:Indigo Carmine; D044602:Cellulases