title: Isolation and characterization of a novel glycosyl hydrolase family 74 (GH74) cellulase from the black goat rumen metagenomic library.
abstract: This study aimed to isolate and characterize a novel cellulolytic enzyme from black goat rumen by using a culture-independent approach. A metagenomic fosmid library was constructed from black goat rumen contents and screened for a novel cellulase. The KG37 gene encoding a protein of 858 amino acid residues (92.7 kDa) was isolated. The deduced protein contained a glycosyl hydrolase family 74 (GH74) domain and showed 77% sequence identity to two endo-1,4-β-glucanases from Fibrobacter succinogenes. The novel GH74 cellulase gene was overexpressed in Escherichia coli, and its protein product was functionally characterized. The recombinant GH74 cellulase showed a broad substrate spectrum. The enzyme exhibited its optimum activity at pH 5.0 and temperature range of 20-50 °C. The enzyme was thermally stable at pH 5.0 and at a temperature of 20-40 °C. The novel GH74 cellulase can be practically exploited to convert lignocellulosic biomass to value-added products in various industrial applications in future.
authors.affiliation: Department of Food Science and Technology, Chung-Ang University, Ansung, 456-756, South Korea.
authors.forename: Yun-Hee
authors.initials: YH
authors.lastname: Song
authors.affiliation: Animal Genomics and Bioinformatics Division, National Institute of Animal Science, Rural Development Administration, Wanju, 565-851, South Korea.
authors.forename: Kyung-Tai
authors.initials: KT
authors.lastname: Lee
authors.affiliation: Department of Food Science and Technology, Chung-Ang University, Ansung, 456-756, South Korea.
authors.forename: Jin-Young
authors.initials: JY
authors.lastname: Baek
authors.affiliation: Department of Food Science and Technology, Chung-Ang University, Ansung, 456-756, South Korea.
authors.forename: Min-Ju
authors.initials: MJ
authors.lastname: Kim
authors.affiliation: Animal Genomics and Bioinformatics Division, National Institute of Animal Science, Rural Development Administration, Wanju, 565-851, South Korea.
authors.forename: Mi-Ra
authors.initials: MR
authors.lastname: Kwon
authors.affiliation: Animal Genomics and Bioinformatics Division, National Institute of Animal Science, Rural Development Administration, Wanju, 565-851, South Korea.
authors.forename: Young-Joo
authors.initials: YJ
authors.lastname: Kim
authors.affiliation: Animal Genomics and Bioinformatics Division, National Institute of Animal Science, Rural Development Administration, Wanju, 565-851, South Korea.
authors.forename: Mi-Rim
authors.initials: MR
authors.lastname: Park
authors.affiliation: Animal Genomics and Bioinformatics Division, National Institute of Animal Science, Rural Development Administration, Wanju, 565-851, South Korea.
authors.forename: Haesu
authors.initials: H
authors.lastname: Ko
authors.affiliation: Department of Biological Sciences, Kyonggi University, Suwon, 442-760, South Korea.
authors.forename: Jin-Sung
authors.initials: JS
authors.lastname: Lee
authors.affiliation: Department of Food Science and Technology, Chung-Ang University, Ansung, 456-756, South Korea. keunsung@cau.ac.kr.
authors.forename: Keun-Sung
authors.initials: KS
authors.lastname: Kim
mesh_terms: D000818:Animals; D002480:Cellulase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D045848:Fibrobacter / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D015723:Gene Library; D005820:Genetic Testing; D006041:Goats / Q000382:microbiology*; D006863:Hydrogen-Ion Concentration; D054892:Metagenome; D056186:Metagenomics; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012417:Rumen / Q000382:microbiology*; D017385:Sequence Homology; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D011994:Recombinant Proteins; D002480:Cellulase