Cellulose binding fusion proteins having a substrate binding reg

Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase

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435 691, 435 6952, 435 697, 435 711, 4351723, 435177, 435179, 435200, 435209, 435803, 530808, 530814, 935 11, 935 14, C12N 914, C12N 1112, C12N 1500, C12P 2100

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052022471

ABSTRACT:
A fusion protein is prepared containing a polypeptide such as an enzyme and an amino acid sequence having a substrate binding region of a polysaccharidase such as cellulase that has essentially no polysaccharidase activity. By contacting the fusion protein with an affinity matrix containing a substrate such as cellulose for the cellulase substrate binding region, the substrate binding region binds to the affinity matrix to immobilize the polypeptide. The polypeptide can be purified by separating the fusion protein or polypeptide from the affinity matrix. The polypeptide can be separated by cleaving the protein with a Cellulomonas fimi protease.

REFERENCES:
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Watanabe, et al., "Gene Cloning of Chitinase Al from Bacillus circulans WL-12 Revealed Its Evolutionary Relationship to Serratia Chitinase and to the Type III Homology Units of Fibronectin," The Journal of Bio. Chem., vol. 265, pp. 15659-15665 (1990).
Watanabe, et al., "Chitinase System of Bacillus circulans WL-12 and Importance of Chitinase A1 in Chitin Degradation," Journal of Bacteriology, vol. 172, pp. 4017-4022 (1990).
Kellett, et al., "Xylanase B and an arabinofuranosidase from Pseudomonas fluorescens subsp. cellulosa contain identical cellulose-binding domains and are encoded by adjacent genes", Biochem. J., vol. 272, pp. 369-376 (1990).
"Purification and Characterization of Endoglucanase C of Cellulomonas fimi, Cloning of the Gene, and Analysis of In Vivo Transcripts of the Gene", by Moser et al. in Applied and Environmental Microbiology, 55:2480-2487 (1989).
"Precise Excision of the Cellulose Binding Domains from Two Cellulomonas fimi Cellulases by a Homologous Protease and the Effect on Catalysis", by Gilkes et al. in The Journal of Biological Chemistry, 263:10401-10407 (1988).
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