Cloning, sequencing, and expression of the gene encoding the Clostridium stercorarium xylanase C in Escherichia coli

• Published in: Bioscience, biotechnology, and biochemistry Vol. 63; no. 9; pp. 1596 - 1604
• Main Authors: Ali, M.K. (Mie Univ., Tsu (Japan). Faculty of Bioresources), Fukumura, M, Sakano, K, Karita, S, Kimura, T, Sakka, K, Ohmiya, K
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 01.09.1999; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; cellulose-binding domain; CLONACION MOLECULAR; CLONAGE MOLECULAIRE; Cloning, Molecular; CLOSTRIDIUM; Clostridium - enzymology; Clostridium - genetics; Clostridium stercorarium; DNA Primers - genetics; DNA, Bacterial - genetics; Endo-1,4-beta Xylanases; Enzyme Stability; ESCHERICHIA COLI; Escherichia coli - genetics; EXPRESION GENICA; EXPRESSION DES GENES; Fundamental and applied biological sciences. Psychology; GENE EXPRESSION; Genes, Bacterial; GENETIC ENGINEERING; Genetics; GENIE GENETIQUE; HIDROLASAS; Hydrogen-Ion Concentration; HYDROLASE; HYDROLASES; Immunochemistry; INGENIERIA GENETICA; Mice; Mission oriented research; MOLECULAR CLONING; Molecular Sequence Data; NUCLEOTIDE SEQUENCE; Open Reading Frames; p-Nitrophenyl-^b-D-cellobioside; p-Nitrophenyl-^b-D-xylopyranoside; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Restriction Mapping; SECUENCIA NUCLEOTIDICA; Sequence Homology, Amino Acid; SEQUENCE NUCLEOTIDIQUE; xylan; xylanase; Xylosidases - chemistry; Xylosidases - genetics; Xylosidases - metabolism; xynC gene; XynC protein; Purification; Nucleotide sequence; Enzyme; Escherichia coli; Sequence alignment; Clostridiales; Homology; Endo-1,4-β-xylanase; Gene expression; Enzymatic activity; Glycosidases; Substrate specificity; Clostridiaceae; Bacteria; Hydrolases; Recombinant protein; Molecular cloning; O-Glycosidases; Molecular domain; Aminoacid sequence; Enterobacteriaceae
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.63.1596

The nucleotide sequence of the Clostridium stercorarium F-9 xynC gene, encoding a xylanase XynC, consists of 3,093 bp and encodes a 1,031-amino acids with a molecular weight of 115,322. XynC is a multidomain enzyme composed of an N-terminal signal peptide and six domains in the following order: two thermostabilizing domains; a family 10 xylanase domain, a family IX cellulose-binding domain, and two S-layer homologous domains. Immunological analysis indicated the presence of XynC in the culture supernatant of C. stercorarium F-9 and in the cells, most likely on the cell surface. XynC purified from a recombinant E. coli was highly active toward xylan and slightly active toward p-nitrophenyl-beta-D-xylopyranoside, p-nitrophenyl-beta-D-cellobioside, p-nitrophenyl-beta-D-glucopyranoside, and carboxymethylcellulose; XynC hydrolyzed xylan and xylooligosaccharides larger than xylotriose to produce xylose and xylobiose. This enzyme was optimally active at 85 degrees C and was stable up to 75 degrees C at pH5.0 and over the pH range of 4 to 7 at 25 degrees C