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Molecular Characterization of the Extracellular poly(3-hydroxyoctanoic Acid) [P(3HO)] Depolymerase Gene of Pseudomonas Fluorescens GK13 and of Its Gene Product

Molecular Characterization of the Extracellular poly(3-hydroxyoctanoic Acid) [P(3HO)] Depolymerase Gene of Pseudomonas Fluorescens GK13 and of Its Gene Product

A Schirmer, D Jendrossek

J Bacteriol. 1994 Nov;176(22):7065-73.

PMID: 7961472

Abstract:

phaZPfi, the gene encoding the extracellular poly(3-hydroxyoctanoic acid) depolymerase of Pseudomonas fluorescens GK13, was cloned, sequenced, and characterized. It comprises 837 bp and is transcribed as a monocistronic message of about 950 bp from a putative sigma 70-like promoter 32 bp upstream of the ATG start codon. The deduced protein of 278 amino acids reveals a typical leader peptide at its N terminus. When expressed in Escherichia coli, the mature depolymerase started with Ala-23, whereas the mature enzyme purified from P. fluorescens GK13 started with both Leu-34 and Arg-35 determining proteins of 26,687 and 26,573 Da, respectively. The depolymerase is a strongly hydrophobic protein and includes the lipase consensus sequence Gly-X-Ser-X-Gly, which is known for serine hydrolases. Replacement of the central residue, Ser-172, in the corresponding sequence (Gly-Ile-Ser-Ser-Gly) of PhaZPfl with alanine resulted in complete loss of enzyme activity, indicating that the poly(3-hydroxyoctanoic acid) depolymerase belongs to the family of serine hydrolases.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP88930089	(±)-3-Hydroxyoctanoic acid		88930-08-9	Price

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