title: Characterization of a gene encoding cellulase from uncultured soil bacteria.
abstract: To detect cellulases encoded by uncultured microorganisms, we constructed metagenomic libraries from Korean soil DNAs. Screenings of the libraries revealed a clone pCM2 that uses carboxymethyl cellulose (CMC) as a sole carbon source. Further analysis of the insert showed two consecutive ORFs (celM2 and xynM2) encoding proteins of 226 and 662 amino acids, respectively. A multiple sequence analysis with the deduced amino acid sequences of celM2 showed 36% sequence identity with cellulase from the Synechococcus sp., while xynM2 had 59% identity to endo-1,4-beta-xylanase A from Cellulomonas pachnodae. The highest enzymatic CMC hydrolysis was observable at pH 4.0 and 45 degrees C with recombinant CelM2 protein. Although the enzyme CelM2 additionally hydrolyzed avicel and xylan, no substrate hydrolysis was observed on oligosaccharides such as cellobiose, pNP-beta-cellobioside, pNP-beta-glucoside, and pNP-beta-xyloside. These results showed that CelM2 is a novel endo-type cellulase.
authors: Microbial Genetics Division, National Institute of Agricultural Biotechnology, Rural Development Administration, Suwon, Korea.
authors: Soo-Jin
authors: SJ
authors: Kim
authors: Chang-Muk
authors: CM
authors: Lee
authors: Bo-Ram
authors: BR
authors: Han
authors: Min-Young
authors: MY
authors: Kim
authors: Yun-Soo
authors: YS
authors: Yeo
authors: Sang-Hong
authors: SH
authors: Yoon
authors: Bon-Sung
authors: BS
authors: Koo
authors: Hong-Ki
authors: HK
authors: Jun
mesh_terms: D000595:Amino Acid Sequence; D001419:Bacteria / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D002266:Carboxymethylcellulose Sodium / Q000378:metabolism; D002480:Cellulase / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015698:Genomic Library; D007723:Korea; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D002480:Cellulase; D002266:Carboxymethylcellulose Sodium