Summary
Endoglucanase C encoded by the celC gene of Clostridium thermocellum has been purified to homogeneity from a recombinant Escherichia coli strain. It was found that this enzyme is highly efficient in degrading glucans with alternating β-1,4- and β-1,3-linkages but lacks activity on unmodified cellulosic substrates. The properties of endoglucanase C were compared to those of Bacillus subtilis β-glucanase, an enzyme used in the brewing industry for β-glucan degradation. Both enzyme cause a rapid decrease of the viscosity of barley β-glucan as a result of internal chain cleavage. Endoglucanase C hydrolyses non-specifically β-1,3- and β-1,4-bonds adjacent to unsubstituted or 4-O′-substituted cellobiose units. Due to its lower pH optimum and increased thermostability endoglucanase C compares favourably with B. subtilis β-glucanase and seems suitable for use in the mashing process of beer brewing.
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References
Aiba S, Kitai K, Imanaka T (1983) Cloning of a thermostable alpha-amylase gene from Bacillus stearothermophilus in Bacillus stearothermophilus and Bacillus subtilis. Appl Environ Microbiol 46:1059–1065
Aspinall GO (1980) Chemistry of cell wall polysaccharides. In: Preiss J (ed) The biochemistry of plants. Vol. 3. Academic Press, New York, pp 473–500
Bamforth CW (1982) Barley β-glucans: their role in malting and brewing. Brewers Digest 1982:22–35
Bamforth CW, Martin HL (1981) β-Glucan and β-glucan solubilase in malting and brewing. J Inst Brew 87:365–371
Borriss R, Zemek J, Augustin J, Bacova Z, Kuniak L (1980) β-1,3-1,4-glucanase in sporeforming microorganisms. II. Production of β-glucan-hydrolases by various Bacillus species. Zbl Bakt II. Abt 135:435–442
Borriss R (1981) Purification and isolation of an extracellular β-glucanase from Bacillus IMET B376. Z Allg Mikrobiol 21:7–17
Cantwell BA, McConnell DJ (1983) Molecular cloning of a Bacillus subtilis β-glucanase gene in Escherichia coli. Gene 23:211–219
Coughlan MP (1985) The properties of fungal and bacterial cellulases with comment on their production and application. Biotechnol Genet Eng Rev 3:39–109
Enari TM, Markkanen PH (1975) Microbial β-glucanases in brewing. Proc Am Soc Brew Chem 33:13–17
Enkenlund J (1972) Externally added β-glucanase. Process Biochem 7:27–29
Hinchliffe E (1984) Cloning and expression of a Bacillus subtilis endo-1,3-1,4-β-d-glucanase gene in Escherichia coli K12. J Gen Microbiol 130:1285–1291
Igarashi O, Noguchi M, Fujimaka M (1968) Studies on the non-starchy polysaccharides of the endosperm of naked berley. V. The degradations of F-1 β-glucan with some cellulases. Agr Biol Chem 32:272–278
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Manners DJ, Wilson G (1976) Purification of malted barley endo-β-d-glucanase by ion exchange chromatography: some properties of an endo-barley-β-d-glucanase. Carbohydrate Res 48:255–264
Moscatelli EA, Ham EA, Rickes EL (1961) Enzymatic properties of a β-glucanase from Bacillus subtilis. J Biol Chem 236:2858–2862
Parrish FW, Perlin AS, Reese ET (1960) Selective enzymolysis of poly-β-d-glucans, and the structure of the polymers. Can J Chem 38:2094–2104
Petre D, Millet J, Longin R, Beguin P, Girard H, Aubert JP (1986) Purification and properties of the endoglucanase C of Clostridium thermocellum produced in Escherichia coli. Biochimie 68:687–695
Reese ET, Mandels M (1966) β-Glucanases other than cellulase. Methods Enzymol 8:607–615
Schwarz W, Bronnenmeier K, Staudenbauer WL (1985) Molecular cloning of Clostridium thermocellum genes involved in β-glucan degradation in bacteriophage lambda. Biotechnol Lett 7:859–864
Schwarz WH, Schimming S, Staudenbauer WL (1987a) Highlevel expression of Clostridium thermocellum cellulase genes in Escherichia coli. Appl Microbiol Biotechnol 27:50–56
Schwarz WH, Bronnenmeier K, Gräbnitz F, Staudenbauer WL (1987b) Activity staining of cellulases in polyacrylamide gels containing mixed linkage β-glucans. Anal Biochem 164:72–77
Sedmark JJ, Grossberg SE (1977) A rapid, sensitive assay for protein using Coomassie brilliant blue G250. Anal Biochem 79:544–552
Soutschek-Bauer E, Staudenbauer WL (1987) Synthesis and secretion of a heat-stable carboxymethylcellulase from Clostridium thermocellum in Bacillus subtilis and Bacillus stearothermophilus. Mol Gen Genet 208:537–541
Suzuki H, Kaneko T (1976) Degradation of barley glucan and lichenan by a Bacillus pumilus enzyme. Agr Biol Chem 40:577–586
Teather RM, Wood PJ (1982) Use of Congo red-polysaccharide interactions in enumeation and characterization of cellulolytic bacteria from bovine rumen. Appl Environ Microbiol 43:777–780
Westermann DH, Huige NJ (1979) Beer brewing. In: Peppler HJ, Perlman D (eds) Microbial Technology. Vol. 2. Academic Press, New York, pp 1–37
Wood TM (1971) The cellulases of Fusarium solani. Purification and specificity of the β-1,4-glucanase and β-glucosidase components. Biochem J 121:353–362
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Schwarz, W.H., Schimming, S. & Staudenbauer, W.L. Degradation of barley β-glucan by endoglucanase C of Clostridium thermocellum . Appl Microbiol Biotechnol 29, 25–31 (1988). https://doi.org/10.1007/BF00258346
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DOI: https://doi.org/10.1007/BF00258346