BBB : Vol. 63 (1999) , No. 9 pp.1596-1604

Cloning, Sequencing, and Expression of the Gene Encoding the Clostridium stercorarium Xylanase C in Escherichia coli

Mursheda K. ALI¹⁾, Masayuki FUKUMURA¹⁾, Katsushi SAKANO¹⁾, Shuichi KARITA²⁾, Tetsuya KIMURA¹⁾, Kazuo SAKKA¹⁾ and Kunio OHMIYA¹⁾

1) Department of Bioscience, Faculty of Bioresources, Mie University
2) Center for Molecular Biology and Genetics, Mie University

(Received April 8, 1999)
(Accepted June 7, 1999)

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-β-D-xylopyranoside, p-nitrophenyl-β-D-cellobioside, p-nitrophenyl-β-D-glucopyranoside, and carboxymethylcellulose. XynC hydrolyzed xylan and xylooligosaccharides larger than xylotriose to produce xylose and xylobiose. This enzyme was optimally active at 85°C and was stable up to 75°C at pH 5.0 and over the pH range of 4 to 7 at 25°C.

Key words:

xylanase; Clostridium stercorarium; cellulose-binding domain

	To cite this article:
	Mursheda K. ALI, Masayuki FUKUMURA, Katsushi SAKANO, Shuichi KARITA, Tetsuya KIMURA, Kazuo SAKKA and Kunio OHMIYA, “Cloning, Sequencing, and Expression of the Gene Encoding the Clostridium stercorarium Xylanase C in Escherichia coli”, Biosci. Biotechnol. Biochem., Vol. 63, 1596-1604 (1999) .

	doi:10.1271/bbb.63.1596
	JOI JST.JSTAGE/bbb/63.1596