Cloning and expression of the Zymomonas mobilis 'production of ethanol' genes in Lactobacillus casei

R. Shane Gold, Michael M. Meagher, Suxiang Tong, Robert W. Hutkins, Tyrrell Conway

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Abstract

This study describes the expression of the Zymomonas mobilis genes coding for pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adh) in Lactobacillus casei 686. To promote transcription, the promoter and ribosome binding site (RBS) from the Lactococcus lactus subsp. lactis-derived vector, pMGE36e, were inserted upstream of the pdc gene. The former sequences were positioned such that translation of pdc was coupled to translation of an 81- base pair open reading frame terminating within the pdc initiation site. The recombinant plasmid (pRSG02) was electroporated into L. casei, and transformants were obtained. Northern analysis confirmed the production of a 3.1-kb transcript corresponding to the predicted size of the PET operon. Western blot analyses revealed that the recombinant strain expressed both enzymes. The recombinant produced more than twice the ethanol produced by the parental L. casei strain.

Original languageEnglish (US)
Pages (from-to)256-260
Number of pages5
JournalCurrent Microbiology
Volume33
Issue number4
DOIs
StatePublished - Oct 1 1996

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ASJC Scopus subject areas

  • Microbiology
  • Applied Microbiology and Biotechnology

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