Nucleotide sequence of the cellulase gene<i>celD</i>encoding endoglucanase D of<i>Clostridium thermocellum</i>

Joliff, Gwennaël; Béguin, Pierre; Aubert, Jean-Paul

doi:10.1093/nar/14.21.8605

Cited by 128 publications

(54 citation statements)

References 25 publications

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“…This assumption is supported by the finding that most functionally important residues identified in the six substrate-binding sites of CelD are conserved in CenC. In this context, it is interesting to note that both CenC (this study) and CelD (10,23,48) are highly active on chromophoric and fluorophoric glycosides derived from ␤-D-1,4-cellobiose and lactose. The absence of significant activity of CenB on these substrates supports the classification of CenC and CenB in different subfamilies (50).…”

Section: Discussionsupporting

confidence: 52%

Characterization of CenC, an enzyme from Cellulomonas fimi with both endo- and exoglucanase activities

et al. 1996

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The cenC gene, encoding ␤-1,4-glucanase C (CenC) from Cellulomonas fimi, was overexpressed in Escherichia coli with a tac-based expression vector. The resulting polypeptide, with an apparent molecular mass of 130 kDa, was purified from the cell extracts by affinity chromatography on cellulose followed by anion-exchange chromatography. N-terminal sequence analysis showed the enzyme to be properly processed. Mature CenC was optimally active at pH 5.0 and 45؇C. The enzyme was extremely active on soluble, fluorophoric, and chromophoric glycosides (4-methylumbelliferyl ␤-glycosides, 2-chloro-4-nitrophenyl-␤-D-cellobioside, and 2-chloro-4-nitrophenyl-lactoside) and efficiently hydrolyzed carboxymethyl cellulose, barley ␤-glucan, lichenan, and, to a lesser extent, glucomannan. CenC also hydrolyzed acid-swollen cellulose, Avicel, and bacterial microcrystalline cellulose. However, degradation of the latter was slow compared with its degradation by CenB, another C. fimi cellulase belonging to the same enzyme family. CenC acted with inversion of configuration at the anomeric carbon, in accordance with its classification as a family 9 member. The enzyme released mainly cellobiose from soluble cellodextrins and insoluble cellulose. Attack appeared to be from the reducing chain ends. Analysis of carboxymethyl cellulose hydrolysis suggests that CenC is a semiprocessive enzyme with both endo-and exoglucanase activities.

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Section: Discussionsupporting

confidence: 52%

Characterization of CenC, an enzyme from Cellulomonas fimi with both endo- and exoglucanase activities

et al. 1996

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“…CelF, the CelS equivalent in C. cellulolyticum, also displays significant activity in reducing the viscosity of a CMC solution (271). Some of the cloned endoglucanase genes are celA (26), celB (105), celC (298), celD (143,326), celE (113), celF (245), celG (188), celH (361), celI (101,123,373), celJ (encoding component S2 of the complex) (1,8), celM (162), celN (373), celQ (9), celT (172), and celX (113).…”

Section: Genes and Enzymesmentioning

confidence: 99%

Cellulase, Clostridia, and Ethanol

Demain

Newcomb

2005

Microbiol Mol Biol Rev

810

579

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SUMMARY Biomass conversion to ethanol as a liquid fuel by the thermophilic and anaerobic clostridia offers a potential partial solution to the problem of the world's dependence on petroleum for energy. Coculture of a cellulolytic strain and a saccharolytic strain of Clostridium on agricultural resources, as well as on urban and industrial cellulosic wastes, is a promising approach to an alternate energy source from an economic viewpoint. This review discusses the need for such a process, the cellulases of clostridia, their presence in extracellular complexes or organelles (the cellulosomes), the binding of the cellulosomes to cellulose and to the cell surface, cellulase genetics, regulation of their synthesis, cocultures, ethanol tolerance, and metabolic pathway engineering for maximizing ethanol yield.

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“…A large number of genes have been identified that code for various endoglucanases (Beguin et al, 1985 ;Grepinet & Beguin, 1986;Joliff et al, 1986; McGavin & Forsberg, Substrate spec8city and enzyme assays. The E. coli periplasmic fraction was used for substrate specificity assays for the cloned enzyme.…”

Section: Introductionmentioning

confidence: 99%

Nucleotide sequence and characteristics of endoglucanase gene engB from Clostridium cellulovorans

Foong

Hamamoto²,

Shoseyov³

et al. 1991

Journal of General Microbiology

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An endoglucanase gene, engB, from ClostridYum cellulovorans, previously cloned into pUCl9, has been further characterized and its product investigated. The enzyme, EngB, encoded by the gene was secreted into the periplasmic space of Escherichiu coli. The enzyme was active against carboxymethylcellulose, xylan and lichenan but not Avicel (crystalline cellulose). The sequenced gene showed an open reading frame of 1323 base pairs and coded for a protein with a molecular mass of 48.6 kDa. The mRNA contained a typical Gram-positive ribosomebinding site sequence GGAGG and a sequence coding for a putative signal peptide. There is high amino acid and base sequence homology between the N-terminal regions of EngB and another C. cellulovorans endoglucanase, EngD, but they differ significantly in their C-termini. Deletion analyses revealed that up to 32 amino acids of the N-terminus and 52 amino acids of the C-terminus were not required for catalytic activity. The conserved reiterated domains at the C-terminus of EngB were similar to those from endoglucanases from other cellulolytic bacteria. According to our deletion analyses, this region is not needed for catalytic activity.

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Nucleotide sequence of the cellulase genecelDencoding endoglucanase D ofClostridium thermocellum

Cited by 128 publications

References 25 publications

Characterization of CenC, an enzyme from Cellulomonas fimi with both endo- and exoglucanase activities

Characterization of CenC, an enzyme from Cellulomonas fimi with both endo- and exoglucanase activities

Cellulase, Clostridia, and Ethanol

Nucleotide sequence and characteristics of endoglucanase gene engB from Clostridium cellulovorans

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