Purification and properties of an endo-1,4-beta-glucanase translated from a Clostridium josui gene in Escherichia coli.
An endoglucanase encoded by a gene of Clostridium josui was expressed in Escherichia coli and purified. The homogeneous enzyme, with a molecular weight of 39,000, revealed maximum endoglucanase activity at pH 7.2 to 7.5 and a temperature of 65 to 70 degrees C. The enzyme was stable at a temperature lower than 45 degrees C (the growth temperature of the bacterium) in the range of pH 4.5 to 9.0. The amino acid sequence of the enzyme at the N terminus was Val-Glu-Glu-Asp-Ser-Ser-His-Leu-Ile-Thr-Asn-Gln-Ala-Lys-Lys----. The enzyme hydrolyzed cellotetraose to cellobiose and then transferred cellobiose to the residual cellotetraose. The resulting cellohexaose was cleaved to cellotriose.
Fujino
T
T
Research and Technological Division, Nagoya Seiraku Co. Ltd., Japan.
Sasaki
T
T
Ohmiya
K
K
Shimizu
S
S
D000595:Amino Acid Sequence; D002475:Cellobiose; D002480:Cellulase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002482:Cellulose / Q000031:analogs & derivatives*; D003013:Clostridium / Q000201:enzymology* / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005798:Genes, Bacterial; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides; D014176:Protein Biosynthesis; D013379:Substrate Specificity; D013696:Temperature; D013780:Tetroses; D014170:Transformation, Genetic
D009844:Oligosaccharides; D013780:Tetroses; D002475:Cellobiose; C048468:cellotetraose; D002482:Cellulose; D002480:Cellulase