Showing papers by "Neil R. Gilkes published in 1989"
TL;DR: The CBDs retain their cellulose-binding properties when fused to heterologous proteins and can be used as affinity tags for protein purification, and for enzyme immobilization.
114 citations
TL;DR: It is believed that the cellulose-binding domain (CBDCex) of Cex offers a simple, inexpensive and widely applicable method of enzyme immobilization.
Abstract: We have constructed a fusion between the gene (abg) for a β-glucosidase (Abg) of an Agrobacterium sp. and part of the gene (cex) for an exoglucanase (Cex) of Cellulomonas fimi. The abg-cex fusion encodes a fusion protein (Abg-CBDCex) with the cellulose-binding properties of Cex and β-glucosidase activity of Abg. The fusion protein retained 42 percent of its β-glucosidase activity when bound to cellulose. The binding was stable enough to allow continuous hydrolysis of substrate by the enzyme without detectable leaching of the enzyme from the cellulose. We believe that the cellulose-binding domain (CBDCex) of Cex offers a simple, inexpensive and widely applicable method of enzyme immobilization.
97 citations
TL;DR: CenA′‐′PhoA fusion polypeptides which contain the entire cellulose‐binding domain of CenA bind to cellulose, allowing their purification from periplasmic extracts in a single, facile step and has implications for purification or immobilisation of chimeric proteins on a cheap cellulose matrix.
78 citations
TL;DR: Results indicated that the catalytic domain adopts a tightly folded conformation affording protection from proteolytic attack, which suggests that CenA has a tertiary structure which resembles that of certain fungal cellulases.
72 citations
TL;DR: Two nonglycosylated endoglucanases which bind to Sephadex were purified from culture supernatants of Cellulomonas fimi grown on microcrystalline cellulose, suggesting that the enzymes were related.
Abstract: Two nonglycosylated endoglucanases which bind to Sephadex were purified from culture supernatants of Cellulomonas fimi grown on microcrystalline cellulose. Their Mrs were 120,000 and 130,000. The N-terminal amino acid sequences of the enzymes were identical, suggesting that the enzymes were related. A DNA fragment encoding this N-terminal sequence was cloned in Escherichia coli. The nucleotide sequence corresponding to the N-terminal amino acid sequence was preceded by a sequence encoding a typical leader peptide. Transcripts hybridizing to the cloned fragment were detected in total RNA isolated from C. fimi cells grown on carboxymethyl cellulose but not from cells grown on glycerol or glucose. Transcription started at a cluster of sites 53 to 59 nucleotides upstream of a GUG translation initiation codon and terminated at either of two closely spaced C residues immediately downstream of a region of potential secondary structure. The size of the transcript was approximately 3.5 kilobases, sufficient to encode a polypeptide of 130 kilodaltons. The 130-kilodalton polypeptide is designated endoglucanase C (CenC), and the gene encoding it is designated cenC.
49 citations
28 Jun 1989
TL;DR: In this paper, novel polypeptide compositions and methods for their use are provided comprising fusion proteins capable of binding to a polysaccharide matrix The compositions may be synthesized or prepared by recombinant DNA technology.
Abstract: Novel polypeptide compositions and methods for their use are provided comprising fusion proteins capable of binding to a polysaccharide matrix The compositions may be synthesized or prepared by recombinant DNA technology
33 citations
01 Jan 1989
2 citations
01 Jan 1989
TL;DR: Etude des enzymes cellulolytiques de cellulomonas fimi, β glucosidases chez Trichoderma, production et purification de xylanases.
Abstract: Etude des enzymes cellulolytiques. Cellulases de cellulomonas fimi, β glucosidases. Systemes enzymatiques chez Trichoderma. Production et purification de xylanases
1 citations
Patent•
28 Jun 1989
TL;DR: The authors decrites de nouvelles compositions polypeptidiques and des procedes pour leur emploi, comprenant des proteines de fusion susceptible de se lier a matrice de polysaccharide.
Abstract: Sont decrites de nouvelles compositions polypeptidiques et des procedes pour leur emploi, comprenant des proteines de fusion susceptibles de se lier a une matrice de polysaccharide. Ces compositions peuvent etre synthetisees ou preparees par des techniques de recombinaison d'ADN.