Expression of ATP Nucleotide 3′-Pyrophosphokinase Gene from Streptomyces in Escherichia Coli and its Effects on Host Cells

Satoshi Ezaki, Takashi Toyono, Shigeru Muta, Jun ichiro Mukai

Research output: Contribution to journalArticle

Abstract

Several actinomycetes produce nucleotide 3′-pyrophosphokinase (EC 2.7.6.4), which tansfers the 5′-β, γ-pyrophosphoryl group of ATP or dATP to a variety of purine and pyrimidine nucleotides at their 3′-OH site. To elucidate the expression mechanism and to attempt the use of a gene coding for this enzyme in a heterologous host, we expressed the gene from Streptomyces morookaensis in Escherichia coli. A DNA fragment that included this gene was inserted downstream of the lac promoter in a multicopy plasmid, pUC 119. The resultant plasmid, pUP3, was introduced into E. coli JM 109. When the cells were incubated in the presence of isopropyl-thio-β-D-galactopyranoside (IPTG), the enzyme was found to be secreted into periplasmic space. E. coli cells harboring pUP 312, derived from pUP 3, grew slowly in the presence of IPTG. 3′-Pyrophosphoryl nucleotides were detected in the nucleotide pools of the E. coli cells that produced this enzyme.

Original languageEnglish
Pages (from-to)695-702
Number of pages8
JournalNippon Nogeikagaku Kaishi
Volume69
Issue number6
DOIs
Publication statusPublished - Jan 1 1995

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All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science
  • Chemistry (miscellaneous)
  • Medicine (miscellaneous)

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