title: Cloning and characterization of a novel GH44 family endoglucanase from mangrove soil metagenomic library.
abstract: A novel endoglucanase gene, mgcel44, was isolated from a mangrove soil metagenomic library by functional-based screening. It encodes a 648-aa peptide with a catalytic domain of glycosyl hydrolase family 44. The deduced amino acid sequence of mgcel44 shares less than 50 % identity with endoglucanases in GenBank database. mgcel44 was cloned and overexpressed in Escherichia coli. The recombinant enzyme, MgCel44, has a molecular mass of 70.8 kDa as determined by SDS-PAGE. Its optimal pH and temperature for activity were 6 and 45 °C, respectively. It was highly active at 25-45 °C and pH 5-8. Its activity was enhanced in 0.5 M NaCl by >1.6-fold and stable up to 1.5 M NaCl. MgCel44 was resistant to several organic solvents and had high activity at 15 % (v/v) solvent after incubating for 24 h at 25 °C.
authors.affiliation: Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, People's Republic of China, maizhimao@163.com.
authors.forename: Zhimao
authors.initials: Z
authors.lastname: Mai
authors.forename: Hongfei
authors.initials: H
authors.lastname: Su
authors.forename: Jian
authors.initials: J
authors.lastname: Yang
authors.forename: Sijun
authors.initials: S
authors.lastname: Huang
authors.forename: Si
authors.initials: S
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D032407:Avicennia; D002480:Cellulase / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability / Q000187:drug effects; D015723:Gene Library; D006863:Hydrogen-Ion Concentration; D007477:Ions; D056186:Metagenomics; D008670:Metals / Q000494:pharmacology; D008969:Molecular Sequence Data; D005810:Multigene Family; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D012987:Soil; D012997:Solvents / Q000494:pharmacology; D013379:Substrate Specificity / Q000187:drug effects; D013696:Temperature
chemical_list: D007477:Ions; D008670:Metals; D011994:Recombinant Proteins; D012987:Soil; D012997:Solvents; D012965:Sodium Chloride; D002480:Cellulase