Separation and characterization of the complexes constituting the cellulolytic enzyme system of Clostridium thermocellum (original) (raw)
Abstract
Clostridium thermocellum, strain JW20 (ATCC 31449) when growing in cellulose produces a cellulolytic enzyme system, that at the early stage of the fermentation is largely bound to the substrate. As cellulose is consumed the bound enzyme is released as free enzyme to the culture fluid. The bound enzyme fraction extracted with distilled water from the cellulose contains two major components, a large complex (Mr≃100×106) and a small complex Mr≃4.5×106) which were separated by gel filtration and sucrose solved by affinity chromatography into a complex that binds to the column and into a non-bindable mixture of proteins. All four fractions have endo-_β_-glucanase activity but only the two bound complexes and the free bindable complex hydrolyze crystalline cellulose with cellobiose as the main product. These three complexes are qualitatively similar in that they each contain about 20 different polypeptides (Mr values from 45,000 to 200,000) of which about ten are major components. However, the relative amounts of some of the peptides in the complexes differ. At least four polypeptides of the complexes have endo-_β_-glucanase activity.
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Abbreviations
CM:
cellulose, carboxymethyl cellulose
CMCase:
carboxymethyl cellulase cosidered endo-_β_-1,4-glucanase
SDS:
sodium dodecyl sulfate
YAS:
yellow affinity substance
YAS-cellulose:
yellow affinity substance-cellulose complex
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Author notes
- Koyu Hon-nami
Present address: Mitsubishi-Kasei Institute of Life Sciences, 11, Minamiooya, Machida-Shi, Tokyo, Japan - Michael P. Coughlan
Present address: Dept. of Biochemistry, University College, Galway, Ireland
Authors and Affiliations
- Center of Biological Resource Recovery, University of Georgia, 30602, Athens, GA, USA
Koyu Hon-nami, Michael P. Coughlan, Hiromi Hon-nami & Lars G. Ljungdahl - Department of Biochemistry, University of Georgia, 30602, Athens, GA, USA
Koyu Hon-nami, Michael P. Coughlan, Hiromi Hon-nami & Lars G. Ljungdahl
Authors
- Koyu Hon-nami
- Michael P. Coughlan
- Hiromi Hon-nami
- Lars G. Ljungdahl
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Hon-nami, K., Coughlan, M.P., Hon-nami, H. et al. Separation and characterization of the complexes constituting the cellulolytic enzyme system of Clostridium thermocellum.Arch. Microbiol. 145, 13–19 (1986). https://doi.org/10.1007/BF00413021
- Received: 03 February 1986
- Accepted: 07 March 1986
- Issue date: June 1986
- DOI: https://doi.org/10.1007/BF00413021