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

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

R V Stern  1 , G T Howard
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The polyester polyurethanase gene (pueA) from Pseudomonas chlororaphis encodes a lipase

R V Stern et al. FEMS Microbiol Lett. .

Abstract

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.

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