A new method for measuring scouring efficiency of natural fibers based on the cellulose-binding domain-beta-glucuronidase fused protein.
Cellulose-binding domains (CBDs) are characterized by their ability to strongly bind to different forms of cellulose. This study examined the use of a recombinant CBD fused to the reporter enzyme beta-glucuronidase (CBD-GUS) to determine the extent of removal of the water-repellent waxy component of cotton fiber cuticles following hydrolytic treatment, i.e., scouring. The CBD-GUS test displayed higher sensitivity and repeatability than conventional water absorb techniques applied in the textile industry. Increases in the levels of CBD-GUS bound to the exposed cellulose correlated to increases in the fabric's hydrophilicity as a function of the severity of the scouring treatment applied, clearly indicating that the amount of bound enzyme increases proportionally with the amount of available binding sites. The binding of CBD-GUS also gave measurable and repeatable results when used on untreated or raw fabrics in comparison with conventional water drop techniques. The quantitative response of the reaction as bound enzyme activity was optimized for fully wettable fabrics. A minimal free enzyme concentration-to-swatch weight ratio of 75:1 was found to be necessary to ensure enzyme saturation (i.e., a linear response), corresponding to a free enzyme-to-bound enzyme ratio of at least 3:5.
Degani
Ofir
O
Department of Environmental Biotechnology, MIGAL-Galilee Technology Center, South Industrial Zone, Kiryat Shmona 10200, Israel.
Gepstein
Shimon
S
Dosoretz
Carlos G
CG
D019677:Artificial Gene Fusion; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism; D002863:Chromogenic Compounds / Q000032:analysis; D032561:Cotton Fiber / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism*; D011133:Polysaccharide-Lyases; D011485:Protein Binding; D017434:Protein Structure, Tertiary / Q000235:genetics; D011549:Pseudomonas / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D002863:Chromogenic Compounds; D011994:Recombinant Proteins; D002482:Cellulose; D002265:Carboxylic Ester Hydrolases; C019952:cutinase; D005966:Glucuronidase; D011133:Polysaccharide-Lyases; C027349:pectin lyase