Purification and properties of an endo-β-1,4-glucanase from Clostridium thermocellum

Pétré, Jean; Longin, R.; Millet, J

doi:10.1016/s0300-9084(81)80061-2

Cited by 132 publications

(73 citation statements)

References 11 publications

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“…Bacterial cellulose (BC) has an average DP of 2,000 -3,000 (Hestrin, 1963;Fierobe et al, 2002;Valjamae et al, 1999), while bacterial microcrystalline cellulose (BMCC) prepared by treatment of BC with acids ranges from 130 -1,300, depending on hydrolysis conditions (Valjamae et al, 1999). The DP of phosphoric-acid swollen cellulose (PASC) ranges from 30 to more than 1,000 Krassig, 1985;Petre et al, 1981;Wood and McCrae, 1972), depending on the DP of the starting substrate (Wood, 1988;Hoshino et al, 1997), as well as the phosphoric acid incubation time and temperature (Krassig, 1993).…”

Section: Degree Of Polymerizationmentioning

confidence: 99%

Toward an aggregated understanding of enzymatic hydrolysis of cellulose: Noncomplexed cellulase systems

Zhang

Lynd

2004

Biotech & Bioengineering

1,640

1,284

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Information pertaining to enzymatic hydrolysis of cellulose by noncomplexed cellulase enzyme systems is reviewed with a particular emphasis on development of aggregated understanding incorporating substrate features in addition to concentration and multiple cellulase components. Topics considered include properties of cellulose, adsorption, cellulose hydrolysis, and quantitative models. A classification scheme is proposed for quantitative models for enzymatic hydrolysis of cellulose based on the number of solubilizing activities and substrate state variables included. We suggest that it is timely to revisit and reinvigorate functional modeling of cellulose hydrolysis, and that this would be highly beneficial if not necessary in order to bring to bear the large volume of information available on cellulase components on the primary applications that motivate interest in the subject. B

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Section: Degree Of Polymerizationmentioning

confidence: 99%

Toward an aggregated understanding of enzymatic hydrolysis of cellulose: Noncomplexed cellulase systems

Zhang

Lynd

2004

Biotech & Bioengineering

1,640

1,284

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“…It is the major endoglucanase in the cellulosome as demonstrated by both early studies (Petre et al 1981) and recent proteomic analysis (Gold and Martin 2007;Zverlov et al 2005). It is optimally active at 75°C and is resistant to the denaturing effects of detergents and organic solvents (Schwarz et al 1986).…”

Section: Introductionmentioning

confidence: 99%

“…The endoglucanase CelA was one of the first cellulases to be purified (Petre et al 1981). It is the major endoglucanase in the cellulosome as demonstrated by both early studies (Petre et al 1981) and recent proteomic analysis (Gold and Martin 2007;Zverlov et al 2005).…”

Section: Introductionmentioning

confidence: 99%

Mutations from a family-shuffling-library reveal amino acid residues responsible for the thermostability of endoglucanase CelA from Clostridium thermocellum

2010

Biotechnol Lett

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We constructed a library of chimeras from the major endoglucanase, CelA, of Clostridium thermocellum and a less stable endoglucanase CelB from Clostridium josui with multiple point mutations using low-fidelity family-shuffling method. Mutations that inactivated the enzyme were rapidly eliminated with high-throughput screening. The activities and thermostabilities of selected variants were evaluated, and four amino acid substitutions, K249R, P258S, S329N and E355G, were identified as having significant impact on the thermostability of CelA without affecting enzymatic activity. In the crystal structure of CelA, most of them are away from the activity cleft and are responsible for the stabilization of secondary structures.

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“…A recent ultrastructural study has confirmed the existence of large extracellular complexes in C. thermocellum and has also provided evidence for the likely mechanism of cellulolysis (Bayer & Lamed, 1986). Biochemical analysis of the cellulosome has achieved some success through the purification and characterization of two endoglucanases with molecular sizes of 56 kDa (Petre et al, 1981) and around 90 kDa (Ng & Zeikus, 1981). The objective of complementary genetic studies has been to dissect and characterize the cellulase complex by cloning and expressing cellulase-encoding genes in a suitable host.…”

Section: Introductionmentioning

confidence: 99%

Molecular Cloning of Clostridium Thermocellum DNA and the Expression of Further Novel Endo- ,4-glucanase Genes in Escherichia Coli

Romaniec

Clarke²,

Hazlewood³

1987

Microbiology

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Sau3A fragments of Clostridium thermocellum (NCIB 10682) DNA were ligated into the BamHI site of pBR322 and expressed in Escherichia coli HBlOl and a Lac-mutant thereof. Twentyeight clones with carboxymethylcellulase (CMCase) activity were selected from two libraries by means of the Congo Red plate assay. Restriction enzyme analysis indicated that the CMCase+ clones contained a total of 13 unique DNA inserts. Hybridization of recombinant plasmids with chromosomal DNA confirmed the physical maps in all but one case and was further used to demonstrate the absence of homology between the Hind111 restriction fragments of similar size which occurred in many of the clones. Without exception, CMCase+ E. coli clones expressed endoglucanase activity, but differed with respect to the amount and nature of the enzyme activity produced; additionally, some clones had exoglucanase activity which, in at least one case, was not attributable to the production of a second enzyme. For a few selected clones, the partially purified CMCase was analysed by electrophoresis. A temperature profile characteristic of a thermostable enzyme was demonstrated for the endoglucanase of one of the most active clones. Based on the evidence presented here, it is probable that the 13 unique DNA fragments described do not contain any of the C. thermocellum endoglucanase genes previously cloned.

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Purification and properties of an endo-β-1,4-glucanase from Clostridium thermocellum

Cited by 132 publications

References 11 publications

Toward an aggregated understanding of enzymatic hydrolysis of cellulose: Noncomplexed cellulase systems

Toward an aggregated understanding of enzymatic hydrolysis of cellulose: Noncomplexed cellulase systems

Mutations from a family-shuffling-library reveal amino acid residues responsible for the thermostability of endoglucanase CelA from Clostridium thermocellum

Molecular Cloning of Clostridium Thermocellum DNA and the Expression of Further Novel Endo- ,4-glucanase Genes in Escherichia Coli

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