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. 1990 Aug;54(8):2039-45.

Cloning and nucleotide sequence of an esterase gene from Pseudomonas fluorescens and expression of the gene in Escherichia coli

K D Choi  1 , G H Jeohn, J S Rhee, O J Yoo
Affiliations
  • PMID: 1368608
Free article

Cloning and nucleotide sequence of an esterase gene from Pseudomonas fluorescens and expression of the gene in Escherichia coli

K D Choi et al. Agric Biol Chem. 1990 Aug.
Free article

Abstract

A gene coding for a novel esterase of Pseudomonas fluorescens was cloned in this study. DNA sequencing showed that the open reading frame is comprised of 708 nucleotides. The coding sequence of the gene is preceded by a potential Shine-Dalgarno sequence and by a promoter-like structure. Following the stop codon a structure reminiscent of the E. coli rho-independent terminator is present. The enzyme expressed in an E. coli clone was mostly in the periplasmic space, released to the outside of the cell by osmotic shock and purified to homogeneity by QAE-Sephadex A-50 and DEAE-Sepharose columns. The native form of the enzyme consisted of two identical subunits, each with a molecular weight of 27,000. By studying the properties and substrate specificity, the enzyme was classified as an arylesterase (EC 3.1.1.2).

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