title: A novel esterase gene cloned from a metagenomic library from neritic sediments of the South China Sea.
abstract: BACKGROUND
  Marine microbes are a large and diverse group, which are exposed to a wide variety of pressure, temperature, salinity, nutrient availability and other environmental conditions. They provide a huge potential source of novel enzymes with unique properties that may be useful in industry and biotechnology. To explore the lipolytic genetic resources in the South China Sea, 23 sediment samples were collected in the depth < 100 m marine areas.
  
  
  RESULTS
  A metagenomic library of South China Sea sediments assemblage in plasmid vector containing about 194 Mb of community DNA was prepared. Screening of a part of the unamplified library resulted in isolation of 15 unique lipolytic clones with the ability to hydrolyze tributyrin. A positive recombinant clone (pNLE1), containing a novel esterase (Est_p1), was successfully expressed in E. coli and purified. In a series of assays, Est_p1 displayed maximal activity at pH 8.57, 40°C, with ρ-Nitrophenyl butyrate (C4) as substrate. Compared to other metagenomic esterases, Est_p1 played a notable role in specificity for substrate C4 (kcat/Km value 11,500 S-1m M-1) and showed no inhibited by phenylmethylsulfonyl fluoride, suggested that the substrate binding pocket was suitable for substrate C4 and the serine active-site residue was buried at the bottom of substrate binding pocket which sheltered by a lid structure.
  
  
  CONCLUSIONS
  Esterase, which specificity towards short chain fatty acids, especially butanoic acid, is commercially available as potent flavoring tools. According the outstanding activity and specificity for substrate C4, Est_p1 has potential application in flavor industries requiring hydrolysis of short chain esters.
authors: State Key Laboratories for Agro-biotechnology and College of Biological Sciences, China Agricultural University, Beijing, P.R. China.
authors: Qing
authors: Q
authors: Peng
authors: Xue
authors: X
authors: Zhang
authors: Meng
authors: M
authors: Shang
authors: Xu
authors: X
authors: Wang
authors: Guili
authors: G
authors: Wang
authors: Bingxue
authors: B
authors: Li
authors: Guohua
authors: G
authors: Guan
authors: Ying
authors: Y
authors: Li
authors: Youshao
authors: Y
authors: Wang
mesh_terms: D000595:Amino Acid Sequence; D001419:Bacteria / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002681:China; D004795:Enzyme Stability; D004950:Esterases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism; D019015:Geologic Sediments / Q000032:analysis / Q000382:microbiology*; D007700:Kinetics; D056186:Metagenomics; D008969:Molecular Sequence Data; D009792:Oceans and Seas; D010802:Phylogeny; D012623:Seawater / Q000382:microbiology*; D016415:Sequence Alignment; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D005227:Fatty Acids; D004950:Esterases