The polyester polyurethanase gene (pueA) from Pseudomonas chlororaphis encodes a lipase

FEMS Microbiol Lett. 2000 Apr 15;185(2):163-8. doi: 10.1111/j.1574-6968.2000.tb09056.x.


A gene (pueA, polyurethane esterase A) encoding an extracellular polyurethanase (PueA) was cloned from Pseudomonas chlororaphis into Escherichia coli. The enzyme secreted from E. coli showed esterase activity when assayed with p-nitrophenyl acetate. Subcloning of a 3. 2-kb SalI-EcoRI fragment into a T7 RNA polymerase expression vector (pT7-6) produced a (35)S-labeled protein of 65 kDa. Nucleotide sequencing of pueA showed an open reading frame encoding a 65-kDa protein of 617 amino acid residues, with the serine hydrolase consensus sequence GXSXG. PueA was over-expressed using the pT7-6 vector transformed into E. coli BL21(DE3) and was purified in one step using Sephadex G-75.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins*
  • Base Sequence
  • Cloning, Molecular
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Gene Library
  • Lipase / chemistry
  • Lipase / genetics*
  • Lipase / metabolism
  • Molecular Sequence Data
  • Open Reading Frames
  • Polyurethanes / metabolism*
  • Pseudomonas / enzymology*
  • Pseudomonas / genetics
  • Sequence Analysis, DNA


  • Bacterial Proteins
  • Polyurethanes
  • pueA protein, Pseudomonas chlororaphis
  • Lipase

Associated data

  • GENBANK/AF069748