title: A novel family VII esterase with industrial potential from compost metagenomic library.
abstract: BACKGROUND
  Among the vast microbial genomic resources now available, most microbes are unculturable in the laboratory. A culture-independent metagenomic approach is a novel technique that circumvents this culture limitation. For the screening of novel lipolytic enzymes, a metagenomic library was constructed from compost, and the clone of estCS2 was selected for lipolytic properties on a tributyrin-containing medium.
  
  
  RESULTS
  The estCS2 sequence encodes a protein of 570 amino acid residues, with a predicted molecular mass of 63 kDa, and based on amino acid identity it most closely matches (45%) the carboxylesterase from Haliangium ochraceum DSM 14365. EstCS2 belong to family VII, according to the lipolytic enzyme classification proposed by Arpigny and Jaeger, and it retains the catalytic triad Ser245-Glu363-His466 that is typical of an α/β hydrolase. The Ser245 residue in the catalytic triad of EstCS2 is located in the consensus active site motif GXSXG. The EstCS2 exhibits strong activity toward p-nitrophenyl caproate (C6), and it is stable up to 60°C with an optimal enzymatic activity at 55°C. The maximal activity is observed at pH 9, and it remains active between pH 6-10. EstCS2 shows remarkable stability in up to 50% (v/v) dimethyl sulfoxide (DMSO) or dimethylformamide (DMF). The enzyme has the ability to cleave sterically hindered esters of tertiary alcohol, as well as to degrade polyurethanes, which are widely used in various industries.
  
  
  CONCLUSIONS
  The high stability of EstCS2 in organic solvents and its activity towards esters of ketoprofen and tertiary alcohols, and in polyurethane suggests that it has potential uses for many applications in biotransformation and bioremediation.
authors.affiliation: Bioindustry and Bioenergy Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Yuseong, Daejeon, Republic of Korea.
authors.forename: Chul-Hyung
authors.initials: CH
authors.lastname: Kang
authors.forename: Ki-Hoon
authors.initials: KH
authors.lastname: Oh
authors.forename: Mi-Hwa
authors.initials: MH
authors.lastname: Lee
authors.forename: Tae-Kwang
authors.initials: TK
authors.lastname: Oh
authors.forename: Bong Hee
authors.initials: BH
authors.lastname: Kim
authors.forename: Jung- Hoon
authors.initials: J
authors.lastname: Yoon
mesh_terms: D000595:Amino Acid Sequence; D004950:Esterases / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007218:Industrial Microbiology; D056186:Metagenomics; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D012988:Soil Microbiology; D013379:Substrate Specificity; D013696:Temperature; D014280:Triglycerides / Q000378:metabolism
chemical_list: D014280:Triglycerides; D004950:Esterases