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. 1986 Jun;51(6):1293–1299. doi: 10.1128/aem.51.6.1293-1299.1986

Properties of a Clostridium thermocellum Endoglucanase Produced in Escherichia coli

Wolfgang H Schwarz 1, Folke Gräbnitz 1, Walter L Staudenbauer 1,*
PMCID: PMC239060  PMID: 16347088

Abstract

A cellulase gene of Clostridium thermocellum was transferred to Escherichia coli by molecular cloning with bacteriophage lambda and plasmid vectors and shown to be indentical with the celA gene. The celA gene product was purified from extracts of plasmid-bearing E. coli cells by heat treatment and chromatography on DEAE-Trisacryl. It was characterized as a thermophilic endo-β-1,4-glucanase, the properties of which closely resemble those of endoglucanase A previously isolated from C. thermocellum supernatants. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis the enzyme purified from E. coli exhibited two protein bands with molecular weights of 49,000 and 52,000. It had a temperature optimum at 75°C and was stable for several hours at 60°C. Endoglucanase activity was optimal between pH 5.5 and 6.5. The enzyme was insensitive against end product inhibition by glucose and cellobiose and remarkably resistant to the denaturing effects of detergents and organic solvents. It was capable of degrading, in addition to cellulosic substrates, glucans with alternating β-1,4 and β-1,3 linkages such as barley β-glucan and lichenan.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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